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Introduction to the Holocaust
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Introduction to the HolocaustNestlé Research Center, Vers-chez-les-Blanc, PO Box 44, CH-1000 Lausanne 26, Switzerland., Search for more papers by this author. Nestlé Research Center, Vers-chez-les-Blanc, PO Box 44, CH-1000 Lausanne 26, Switzerland., Search for more papers by this author. Nestlé Research Center, Vers-chez-les-Blanc, PO Box 44, CH-1000 Lausanne 26, Switzerland., Search for Essay more papers by Millennium Development and Jamaica, this author. Francesca Stingele. E-mail firstname.lastname@example.org; Tel. (+41) 21 785 8923; Fax (+41) 21 785 8925. Streptococcus thermophilus Sfi6 produces an exopolysaccharide (EPS) composed of Essay, glucose, galactose and N -acetylgalactosamine in Essay Young the molar ratio of 1:2:1. The Holocaust Essay. The genes responsible for the EPS biosynthesis have been isolated previously and found to plants grow in the, be clustered in a 14.5 kb region encoding 13 genes. The Holocaust Essay. Transfer of this gene cluster into a worn path pdf a non-EPS-producing heterologous host, Lactococcus lactis MG1363, yielded an EPS with a similar high molecular weight, but a different structure from the EPS from the native host. The structure of the recombinant EPS was determined by means of 1 H homonuclear and 1 H- 13 C heteronuclear two-dimensional nuclear magnetic resonance (NMR) spectra and was found to be → 3)-β- d -Glc p -(1 → 3)-α- d -Gal p -(1 → 3)-β- d -Gal p -(1 → as opposed to → 3)[α- d -Gal p -(1 → 6)]-β- d -Glc p -(1 → 3)-α- d -Gal p NAc-(1 → 3)-β- d -Gal p -(1 → for the wild-type S. thermophilus Sfi6. Furthermore, L. lactis MG1363 (pFS101) was also lacking a UDP- N -acetylglucosamine C4-epimerase activity, which would provide UDP-GalNAc for Essay a GalNAc incorporation into the EPS and probably caused the substitution of GalNAc by Gal in the recombinant EPS. This modification implies that (i) bacterial glycosyltransferases could potentially have multiple specificities for that grow in the the donor and the acceptor sugar molecule; and (ii) the repeating unit polymerase can recognize and polymerize a repeating unit that differs in the backbone as well as in the side-chain from its native substrate. In recent years, exopolysaccharides (EPSs) produced by The Holocaust Essay, lactic acid bacteria (LAB) have attracted the attention of the food industry because of their interesting rheological properties combined with their GRAS (generally recognized as safe) status (Sandford and Bard, 1983; Sutherland, 1998). Art Essay. Furthermore, it is Essay now accepted that complex carbohydrates are implicated in a multitude of biological recognition reactions leading, for about and Oversized example, to the modulation of the immune system and host–pathogen interactions (Roberts, 1995; Sutherland, 1998). Numerous such activities have been reported for glycoconjugates and oligosaccharides (Ding et al ., 1995; Zopf and Roth, 1996). However, the same oligosaccharides lined up on a polysaccharide backbone, such as a bacterial EPS, should have an even stronger affinity for its ligands, because of the multivalency offered by the repeating structure. In more general studies, polysaccharides have been reported to exhibit health-beneficial properties, such as antiulcer activities (Nagaoka et al ., 1994) and immune stimulation (Oda et al ., 1982; Nakajima et al ., 1995). Detailed structural studies have been performed on EPSs from different LAB, such as Streptococcus thermophilus (Doco et al ., 1990; Bubb et al ., 1997; Lemoine et al ., 1997; Faber et al ., 1998), Lactococcus lactis ssp. cremoris (Gruter et al ., 1992; Nakajima et al ., 1992), Lactobacillus delbrückii ssp. Essay. bulgaricus (Gruter et al ., 1993), Lactobacillus helveticus (Yamamoto et al ., 1994; 1995; Robijn et al ., 1995a; Staaf et al ., 1996; Stingele et al ., 1997), Lactobacillus sake (Robijn et al ., 1995b), Lactobacillus paracasei (Robijn et al ., 1996a), and difference and secondary sources Lactobacillus acidophilus (Robijn et al ., 1996b). As indicated by the structural diversity of Essay, EPSs produced by LAB, these bacteria contain a vast pool of glycosyltransferases that have not been exploited so far. For this reason, LAB are good model systems for investigating the plants dark complexity of EPS biosynthesis in order to be able to engineer tailor-made polysaccharides with the desired properties. We have previously isolated and characterized the 14.5 kb gene cluster responsible for EPS production from S. thermophilus Sfi6 (Stingele et al ., 1996). According to Essay, homology searches, the 13 genes could be organized into four functional regions (Fig. 1): a central region with four genes showing homology with glycosyltransferases required for the biosynthesis of the EPS-repeating unit ( epsE to epsI ); two regions flanking this central region that showed homology to and Oversized, enzymes involved in polymerization and Essay export ( epsC and epsD for difference primary chain-length control, epsK and epsM for the export and epsJ for the polymerization of the EPS-repeating unit) and, at the beginning of the gene cluster, a gene of which the predicted protein shared homology with a regulatory protein ( epsA ). . Organization of the exopolysaccharide ( eps ) gene cluster from S. thermophilus Sfi6. Flag and hairpin indicate the eps gene cluster promoter and terminator respectively. Essay. Genes are grouped according to Millennium Development Essay, the homologies of their predicted proteins, with proteins of known functions that are a regulatory protein (horizontal hatching), polysaccharide polymerase and chain-length determinator (vertical hatching) and glycosyltransferases (cross-hatching). In order to confirm that this gene cluster alone was capable of directing EPS biosynthesis, it was cloned into the non-EPS-producing heterologous host L. Essay. lactis MG1363, and its capacity to produce EPS was tested on a plate assay. From the plate assay and preliminary fermentation trials carried out with L. lactis (pFS101), it was evident that this recombinant organism was capable of producing EPS (Stingele et al ., 1996). On the other hand, we noted that L. A Worn Path Pdf. lactis (pFS101) had no UDP- N -acetylglucosamine C4-epimerase activity and The Holocaust was therefore unable to produce UDP-GalNAc, which is a worn path pdf required for The Holocaust the incorporation of GalNAc into the S. thermophilus Sfi6 EPS. In this study, we present the characterization and structural determination of the recombinant EPS and show that it is between primary and secondary sources different from the original one produced by The Holocaust Essay, S. A Worn Path Pdf. thermophilus Sfi6. This finding constitutes a first example for heterologous expression of a modified EPS in a Gram-positive microorganism. Characterization of recombinant, EPS-producing L. lactis MG1363 (pFS101) The construction of Essay, strain L. lactis (pFS101) was as described previously (Stingele et al ., 1996). In order to gain information about the stability and EPS-producing capacities of the strain in fermentations, the difference and secondary transcription of the eps gene cluster in The Holocaust S. A Worn Path Pdf. thermophilus Sfi6 was analysed and compared with that of L. lactis (pFS101). As shown in 2Fig. 2A, in S. Essay. thermophilus , the expression of the eps genes is directed by one transcript of ≈15 kb containing the entire cluster. The transcription site has been mapped upstream of epsA by primer extension (Fig. 2B), and the same transcriptional start point was found to be used in L. lactis (pFS101). Therefore, L. lactis (pFS101) recognizes the primary same promoter upstream of epsA as S. thermophilus . . Transcription analysis of the eps gene cluster. The Holocaust. A. Northern hybridization of S. thermophilus Sfi6 and a non-ropy control strain YS4. A Worn Path Pdf. Total RNA was hybridized with PCR-amplified and Essay [α- 32 P]-CTP-labelled fragments of epsA to a worn path pdf, epsB (nucleotides 916–3017 of The Holocaust Essay, accession no. U40830) , epsF to epsG (nucleotides 5876–7532 of accession no. U40830) and epsM (nucleotides 12684–13843 of accession no. Difference Between Primary. U40830). B. Mapping of Essay, transcription start site of the eps gene cluster expressed in Essay Young and Oversized S. thermophilus Sfi6 and L. The Holocaust Essay. lactis MG1363 (pFS101). The proposed promoter sequence upstream of epsA in S. Digital Vs. Traditional. thermophilus Sfi6 was reported by Essay, Stingele and between Mollet (1996). In small-scale batch culture as well as during 24 h fermentations, the growth of L. lactis (pFS101) was similar to that of its untransformed isotype. Additionally, the stability of plasmid pFS101 during fermentation was tested with the ruthenium red plate assay (Stingele et al ., 1996). After 24 h fermentations, only white colonies appeared, showing that the EPS-producing phenotype and thus the plasmid were stably maintained. Hence, plasmid pFS101 and the expression of the eps genes did not seem to interfere with L. lactis house-keeping functions. The Holocaust. The EPS yield after a 24 h fermentation and acetone precipitation of the Development Goals and Jamaica Essay protein-free culture supernatant was only 10 mg l −1 for L. The Holocaust. lactis (pFS101) compared with 100 mg ml −1 for S. thermophilus Sfi6. Estimation of the and secondary molecular weight by gel filtration chromatography on Superose-6 indicated that the recombinant EPS had a molecular weight of ≥ 2 × 10 6 , which is similar to that of the wild-type EPS produced by the S. thermophilus strain. Essay. Therefore, L. lactis (pFS101) was able to produce a high-molecular-mass EPS. UDP -N -acetylglucosamine 4-epimerase activity in S. thermophilus Sfi6 and that dark L. lactis MG1363 strains. The biosynthesis of polysaccharides is initiated by the synthesis of the component sugar nucleotides, which are the donor substrate of the The Holocaust glycosyltransferases. For the polysaccharide components Gal and Glc, the of the Races Essay sugar nucleotides UDP-Gal and UDP-Glc are readily available, as they are required for house-keeping functions. On the other hand, UDP-GalNAc is The Holocaust not essential and has to be synthesized from UDP-GlcNAc by a UDP- N -acetylglucosamine 4-epimerase (EC 22.214.171.124). The presence of this enzyme will thus determine whether UDP-GalNAc is being produced. As the EPS of plants that grow in the dark, S. thermophilus contains GalNAc, the UDP- N -acetylglucosamine 4-epimerase activity of the host L. lactis MG1363 was determined and compared with the level of enzyme activity found in S. thermophilus Sfi6. As anticipated, the specific activity for The Holocaust Essay S. thermophilus was high (1.70 × 10 −1 U mg −1 protein) and corresponded to difference and secondary, a 75.5% reversible conversion rate of The Holocaust Essay, UDP-GalNAc into UDP-GlcNAc. Surprisingly, the activity in L. lactis was hardly detectable (3.8 × 10 −3 U mg −1 protein, conversion rate of ≤ 2.5%) and not detectable for L. Plants Grow In The. lactis (pFS101). Essay. These results indicate that L. lactis (pFS101) is unable to produce UDP-GalNAc from UDP-GlcNAc. Essay About Young. Therefore, L. lactis (pFS101) was either producing an altered EPS without GalNAc or was capable of synthesizing UDP-GalNAc by a novel pathway. In order to gain further insight into The Holocaust the biosynthesis of the about recombinant EPS in the heterologous organism, knowledge of its primary structure was a prerequisite. Composition and NMR spectroscopy of the EPS produced by The Holocaust Essay, L. lactis MG1363 (pFS101) Quantitative monosaccharide analysis combined with the absolute configuration determination showed that the L. lactis (pFS101)-produced EPS contained d -Gal and d -Glc in a molar ratio of 2:1. Methylation analysis of the L. lactis (pFS101)-produced EPS indicated a linear repeating unit with 3-substituted galactopyranose and 3-substituted glucopyranose (molar ratio 2:1). Three anomeric proton resonances could be distinguished in the one-dimensional 1 H NMR spectrum of the EPS of L. Difference And Secondary Sources. lactis (pFS101), namely a low-field H-1 resonance at 5.17 ppm (broad singlet, integral ≈1) and Essay two overlapping H-1 signals at 4.72 ppm (broad singlet, integral ≈2) (Fig. Essay And Oversized. 3). The one-dimensional 1 H NMR spectrum showed broad lines resulting from the The Holocaust high viscosity of the solution observed even at 67°C. The undecoupled 1 H- 13 C heteronuclear single-quantum coherence (HSQC) spectrum was also consistent with the presence of three monosaccharide units, exhibiting two pairs of signals in the anomeric region with integrals ≈1:2 (Fig. 4). . 1 H NMR spectrum of L. lactis MG1363 (pFS101) EPS recorded in 2 H 2 O at 500 MHz and Essay Young 67°C. The protons reported in the spectrum are identified by Essay, the residue letter code (A–C) and the corresponding carbon atom number (1–6). Essay About. The intensity of the proton signals surrounding the residual water signal are decreased compared with the anomeric protons because of presaturation. . Undecoupled 1 H- 13 C HSQC spectrum of L. lactis MG1363 (pFS101) EPS recorded in 2 H 2 O at Essay 67°C. Labelling as in Fig. 3. The three hexopyranose components of the Digital vs. Traditional Art Essay repeating unit were designated A, B, and C in accordance with the decreasing anomeric proton chemical shifts. The anomeric 1 H chemical shift from unit A (5.17 ppm) suggested an α configuration, while the anomeric 1 H chemical shift from The Holocaust Essay units B and C (overlapping at 4.72 ppm) suggested β configurations. No N -acetyl group resonances were observed. The assignment of the 1 H and 13 C NMR signals of the EPS was based on two-dimensional homonuclear (total correlation spectroscopy; TOCSY) and heteronuclear ( 1 H- 13 C HSQC) spectra recorded at 67°C, and the results are shown in Tables 1 and 2 respectively. The 1 H assignment was started from the anomeric resonances of residues A, B and C. By using TOCSY experiments with increasing mixing times, connectivities from the anomeric resonances to the resonances of H-2,3,4 were traced for Essay about Young all three residues. Connectivities to H-5 of residue B and to H-5,6a,6b of residue A were also found. An example of a 85 ms mixing time TOCSY is Essay shown in Fig. That Dark. 5. Partial confirmation of these resonances was obtained by correlating the 1 H resonances with the corresponding 13 C resonances in the undecoupled 1 H- 13 C HSQC spectrum and Essay simultaneously assigning the 13 C chemical shifts. The missing H-5 and H-6a,6b resonances were detected by both finding the C-5 and difference primary sources C-6 resonances in the undecoupled 1 H- 13 C HSQC spectrum and The Holocaust Essay following H-2,3,4 traces in the TOCSY spectra. . 1 H-1H TOCSY spectrum of L. Art Essay. lactis MG 1363 (pFS101) EPS recorded in 2 H 2 O at 67°C with a mixing time of 85 ms. Labelling as in Fig. 3. Low-field H-1 and H-4 chemical shifts (δ H-1 5.17, δ H-4 4.26), a large one-bond 1 H- 13 C scalar coupling ( 1 J C-1,H-1 = 175 Hz), a high-field C-1 chemical shift (98.65 ppm) and the very weak TOCSY cross-peaks between H-1 and The Holocaust both H-5 and Digital vs. Traditional H-6a,6b characterize residue A as being an α- d -Gal p unit. The anomeric chemical shifts (δ H-1 4.72, δ C-1 106.80) and the presence of TOCSY cross-peaks between H-1 and H-5 indicate that residue B must be a β- d -Glc p unit. The anomeric chemical shifts (δ H-1 4.72, δ C-1 106.80), a low-field shifted H-4 (4.18 ppm) incompatible with a β- d -Glc p unit, the C-2 and C-3 chemical shifts (δ C-2 72.62, δ C-3 80.31) more typical of a β- d -Gal p than of a β- d -Glc p residue (Bock and Thøgersen, 1982), a high-field shifted C-4 (67.75 ppm) and the absence of TOCSY cross-peaks between H-1 and either H-5 or H-6a,6b characterize residue C as being a β- d -Gal p unit. The substitution pattern of the three residues was deduced from the Essay 13 C chemical shifts. The low-field shifted C-3 chemical shifts for all three residues [δ C-3 (A) 82.05, δ C-3 (B) 87.71, δ C-3 (C) 80.31] when compared with the a worn path pdf corresponding methyl aldohexopyranoside [δ C-3 (Me α- d -Gal p ) 70.5, δ C-3 (Me β- d -Glc p ) 76.8, δ C-3 (Me β- d -Gal p ) 73.8; Bock et al ., 1983] indicate that all residues are 3-substituted. The sequence of the monosaccharide residues was deduced from the Essay presence of cross-peaks in the heteronuclear multiple-bond correlations (HMBC) spectrum correlating C-1(A) with H-3(C), C-3(C) with H-1(A), C-1(B) with H-3(A), C-3(A) with H-1(B), C-1(C) with H-3(B) and C-3(B) with H-1(C), as well as cross-peaks in the nuclear Overhauser effect (NOESY) spectra between H-1(A) and H-3(C), H-1(B) and H-3(A) and H-1(C) and H-3(B) (data not shown). In conclusion, based on the monosaccharide analysis, the methylation analysis and The Laws Races Essay the various NMR studies, the sequence of the repeating unit of L. Essay. lactis (pFS101) EPS could be formulated as Structure I. In order to facilitate comparisons, the hexopyranoses of the S. thermophilus Sfi6 EPS (Doco et al ., 1990) were designated A to D by labelling the units in the backbone as in L. lactis (pFS101) and the α- d -Gal p side-chain as D. From our biochemical data on the EPS biosynthesis in S. thermophilus (F. Stingele, unpublished), we know that the synthesis of the EPS repeating unit is initiated by the transfer of the β- d -Gal p C residue onto a lipophilic carrier. Consequently, the structure originally published by Doco et al . (1990) is shown as Structure II. The one-dimensional 1 H NMR spectrum of the L. lactis (pFS101) EPS recorded at 67°C was compared with a one-dimensional 1 H NMR spectrum of the Digital vs. Traditional S. thermophilus EPS recorded under identical conditions. The absence of N -acetyl peaks and the presence of three versus four anomeric resonances reflects the changes found in the repeating unit of the two EPSs. As only partial assignments were presented in the original work by Doco et al . (1990), a new NMR analysis of the The Holocaust Essay S. Plants That Grow In The. thermophilus Sfi6 EPS was carried out at The Holocaust Essay 80°C, and the resulting complete assignment is included in Tables 1 and Millennium Development Essay 2 (J. Lemoine, personal communication). Although the chemical shifts were recorded at different temperatures and calibrated differently, some comparisons can be made between the chemical shifts of the The Holocaust two EPSs. For residue A, the α- d -Gal p NAc is Development and Jamaica replaced by an α- d -Gal p residue. The corresponding chemical shifts are very similar, except for the 2 position, which is expected to be different in N -acetylgalactosamine and galactose. For residue B, the Essay absence of a branching point at the 6 position changes the H-6 and C-6 chemical shifts significantly more than the other chemical shifts. Finally, for residue C, which is the only unit to be totally identical between the two EPSs (3-substituted β- d -Gal p ), the overall changes in chemical shifts are small compared with the differences found for residues A and B. We have previously identified and characterized a gene cluster involved in EPS biosynthesis from S. thermophilus Sfi6. The 13 genes responsible for EPS production were cloned into a Gram-positive vector and between and secondary transferred successfully into the non-ropy heterologous host L. lactis MG1363. The resulting strain, L. lactis (pFS101), was capable of producing an EPS, of which the The Holocaust primary structure proved to be different from the one produced by Digital Art Essay, the original S. The Holocaust. thermophilus strain (structures I and Essay about Young and Oversized II respectively). The Holocaust. The first change was that the GalNAc residue in the backbone of the repeating unit of the a worn path pdf S. thermophilus EPS was substituted by a Gal residue in The Holocaust Essay the L. lactis (pFS101) EPS. As the α-1,3- N -acetylgalactosaminyltransferase gene is Young present and The Holocaust Essay functional in the eps gene cluster (F. Stingele, unpublished) and the transcription of the eps genes is efficient (Fig. 2), the expression of the α-1,3- N -acetylgalactosaminyltransferase should also be effective in L. lactis (pFS101). On the other hand, in L. Between Primary And Secondary Sources. lactis (pFS101), no UDP-GlcNAc 4-epimerase activity was detected compared with that in S. thermophilus (75.5% conversion rate), indicating that L. lactis (pFS101) is unable to produce UDP-GalNAc. As no gene involved in the nucleotide sugar pathway is Essay present in the S. The Laws Of The Essay. thermophilus Sfi6 gene cluster, all necessary nucleotide sugars have to be provided by the host. Thus, as L. lactis MG1363 does not seem to provide UDP-GalNAc, it is conceivable that the α-1,3- N -acetylgalactosaminyltransferase could be using UDP-Gal, which would result in a Gal and not a GalNAc incorporation. A donor specificity for both nucleotide sugars, UDP-Gal and UDP-GalNAc, has been reported previously for the mammalian β-1,4-galactosyltransferase by Essay, Palcic and Hindsgaul (1991) and shows that glycosyltransferases do have the potential to transfer Gal and GalNAc. The second modification consisted of difference between primary and secondary sources, a conversion of the tetrameric branched repeating unit with Gal as an α-1,6-linked side-chain ( S. thermophilus ) to Essay, a trimeric linear repeating unit [ L. lactis (pFS101)]. An explanation for the absence of the Gal branching could be that the S. thermophilus α-1,6-galactosyltransferase relies on the presence of the N -acetyl group of the neighbouring unit in order to be functional. This specificity would be reminiscent of the recognition pattern that has been observed for the rat liver α-2,6-sialyltransferase that does not tolerate a modification of the amide grouping of the Galβ(1–4)GlcNAc acceptor (Wlasichuk et al ., 1993). Interestingly though, the β-1,3-glucosyltransferase does not seem to recognize the N -acetyl group of the GalNAc residue in the S. Between Primary. thermophilus EPS, as Glc is transferred to Gal in the L. lactis (pFS101) EPS. Taken together, our evidence combined with what is known about donor and acceptor specificity of Essay, mammalian glycosyltransferases suggests that the modification from the S. thermophilus Sfi6 to the L. lactis MG1363 (pFS101) EPS is primarily caused by the inability of L. lactis MG1363 to Essay Young and Oversized, provide UDP-GalNAc. Essay. This could entail a Gal instead of a GalNAc incorporation for a worn path pdf the second residue of the EPS precursor and no transfer of the The Holocaust Essay Gal residue onto the C-6 of the difference and secondary terminal Glc of the trimeric EPS precursor because of the missing amide group of the second Gal residue. It is tempting to assume that the examples of sugar specificity patterns found among mammalian glycosyltransferases could explain the alteration that occurred in the EPS produced by L. lactis (pFS101), but the sugar recognition pattern for the S. Essay. thermophilus Sfi6 glycosyltransferases remains to be established. An important conclusion resulting from these modifications is that the putative polymerase (EpsJ) and chain-length determinator (EpsC, EpsD) involved in the EPS biosynthesis do not seem to have an exclusive selectivity for the native repeating unit. They are capable of polymerizing both the native tetrameric branched and the recombinant trimeric linear repeating unit to a high molecular weight. This recombinant trimeric repeating unit not only misses the side-chain but also bears a modified monosaccharide in The Laws the backbone (GalNAc to Gal conversion). The Holocaust Essay. Among the numerous examples of polysaccharide synthesis from difference between sources other microorganisms as different as capsule biosynthesis in Streptococcus pneumoniae (Kolkman, 1997) or xanthan biosynthesis in Essay Xanthomonas campestris (Betlach et al ., 1997), these are the first results showing an infidelity of the polymerase towards the polysaccharide backbone. Depending on the genetic background in which a polysaccharide gene cluster is Goals and Jamaica Essay being expressed, this infidelity provides a further degree of flexibility in creating an The Holocaust Essay even larger structural diversity of polysaccharides. The bacterial strains used in this study are listed in that in the Table 3. S. thermophilus Sfi6 or the non-ropy control strain S. thermophilus YS4 were grown routinely in M17 (Difco Laboratories) supplemented with 1% lactose at 42°C. The Holocaust Essay. L. lactis MG1363 (pFS101) was grown in difference primary and secondary M17 supplemented with 1% glucose and 5 μg ml −1 erythromycin at 30°C. Total RNA was isolated from S. thermophilus Sfi6 and YS4 and L. lactis MG1363 (pFS101) by a method modified according to Shaw and The Holocaust Essay Clewell (1985). Exponentially growing cells (20 ml) were poured onto 20 ml of frozen and crushed killing medium (0.02 M Tris-HCl, pH 7.3, 5 mM MgCl 2 , 0.02 M NaN 3 , 400 μg ml −1 chloramphenicol) and centrifuged. The cells were resuspended in 2 ml of lysis buffer (0.02 M Tris-HCl, pH 8, 3 mM EDTA, 0.2 M NaCl) supplemented with 5 mg ml −1 lysozyme and 5 mM EDTA, pH 8, and between primary incubated at The Holocaust Essay 37°C for difference 15 min. After centrifugation, the Essay cells were resuspended in 300 μl of lysis buffer, and 300 μl of lysis buffer supplemented with 2% SDS prewarmed at 95°C were added. The lysate was incubated at 95°C for Digital Art Essay 1 min and, after the addition of Essay, 600 μl of Millennium Development Goals and Jamaica Essay, a phenol–chloroform mixture (1:1), saturated in lysis buffer for a further 3 min at 65°C. The Holocaust Essay. After centrifugation, the supernatant was extracted with chloroform, and the RNA was precipitated from the Digital supernatant with a one-ninth volume of 3 M sodium acetate and two volumes of ethanol and washed with 70% ethanol. Northern blot hybridization was performed as described by Ausubel et al . (1994). Primer extensions were performed with the ‘AMV-Primer extension’ kit (Promega) using 50 μg of Essay, RNA and [γ- 33 P]-ATP-labelled primers according to the supplier's instructions. The sequence of the oligonucleotide used for primer extension mapping of epsA was 5′-CCCAACGTTGACCATCCCCCACG-3′. Enzymatic assay for UDP -N -acetylglucosamine 4-epimerase (EC 126.96.36.199.) Crude cytoplasmic extracts of S. thermophilus Sfi6, L. lactis MG1363 and L. lactis MG1363 (pFS101) were assayed for UDP- N -acetylglucosamine 4-epimerase activity. Cultures (20 ml) grown to an OD 600 of Essay about Young and Oversized, 1.0 were harvested and resuspended in 1 ml of 50 mM potassium phosphate buffer, pH 7.0. One gram of acid-washed glass beads (Sigma) was added to Essay, the tube, and the cells were disrupted by agitating the tubes three times for Digital 3 min with a Mini Bead Beater (Biospec Products). The glass beads and the cell debris were spun down at 10 000 g for 10 min at 4°C, and the supernatant was transferred to The Holocaust, a clean tube. The protein content of the enzymatic extract was determined with the Bradford kit (Bio-Rad). The UDP- N -acetylglucosamine 4-epimerase assay was performed as described by Estrela et al . (1991), with 100 μl of enzyme extract in a final reaction volume of 200 μl. The assay measures the conversion of The Laws Races, UDP-GlcNAc to UDP-GalNAc, because the Essay detection of UDP-GalNAc is that grow in the dark more sensitive. Activities were measured in triplicate and had a SD of < 10%. One unit of The Holocaust Essay, enzyme is defined as the amount that catalyses the formation of 1 μmol of about and Oversized, UDP-GalNAc under standard assay conditions. EPS isolation and characterization. Exopolysaccharides were produced by The Holocaust, continuous fermentation of Young and Oversized, S. thermophilus Sfi6 or L. lactis MG1363 (pFS101) in 10% reconstituted skim milk supplemented with a mixture of amino acids in quantities corresponding to those found in 1% Proteose Peptone No. 3 (Oxoid). The Holocaust Essay. For L. lactis MG1363 (pFS101), 1% glucose and 5 μg ml −1 erythromycin were also added to the growth medium. Fermentations were carried out in a 1 l scale fermenter with a magnetic stirrer (60 rpm) for 24 h at 42°C ( S. thermophilus ) or at 30°C ( L. lactis ) and regulated at pH 5.5 with 2 M NaOH. The EPSs were extracted as follows. Proteins were removed from the spent culture with an Digital vs. Traditional equal volume of trichloroacetic acid (40%). After centrifugation, an equal volume of acetone was added to the supernatant in order to The Holocaust Essay, precipitate the EPS. The precipitated EPS was recovered by centrifugation, redissolved in water and the solution adjusted to pH 7.0 before dialysis against Millennium and Jamaica water for The Holocaust 24 h. Insoluble material was removed by ultracentrifugation, and the EPS was purified further by gel-permeation chromatography on a Superose-6 HR-6 column (Pharmacia). The total neutral sugar content was determined by the phenol–sulphuric acid method (Dubois et al ., 1956). The molecular weight of the EPSs was estimated by gel-permeation chromatography using a Superose-6 HR-6 column connected to Digital vs. Traditional, a fast protein liquid chromatography (FPLC) system (Pharmacia), previously calibrated with commercially available dextrans (Sigma). Chemical analysis of EPS of The Holocaust, L. lactis MG1363 (pFS101) The monosaccharide composition of the purified EPS was estimated by gas-liquid chromatography (GLC) of O -methyloxime acetates (Neeser and Schweizer, 1984) and trimethylsilylated methyl glycosides (Chaplin, 1982; Kamerling and Vliegenthart, 1989). The EPS (0.5 mg) was methanolysed with 0.5 ml of 1 M hydrochloric acid in methanol (24 h at a worn path pdf 85°C), and the solution was neutralized with silver carbonate. The precipitate was removed by Essay, centrifugation, and the supernatant was dried by evaporation under reduced pressure. Trimethylsilylation of the Goals and Jamaica methyl glycosides mixture was carried out by the addition of Essay, 50 μl of fresh silylating reagent (pyridine–chlorotrimethylsilane–1,1,1,3,3,3-hexamethyldisilazane; 5:1:1) 30 min before GLC analysis on a Chrompack CP9002 gas chromatograph equipped with a CP-Sil 43B fused silica capillary column (25 m × 0.32 mm) using a temperature programme from Digital vs. Traditional 130°C to 230°C at 4°C min −1 . The absolute configuration of the monosaccharides was determined according to the methods of Gerwig et al . (1978; 1979). After methanolysis carried out in the same way as for the monosaccharide composition analysis, the Essay dried sample was dissolved in 100 μl of 1 M (-)-2-butanolic hydrochloric acid and The Laws of the Essay heated for 8 h at 85°C. Essay. After neutralization with silver carbonate, the difference and secondary precipitate was removed by centrifugation, and Essay the supernatant was dried by evaporation under reduced pressure. Subsequent trimethylsilylation and GLC analysis were performed as for difference between sources the monosaccharide composition analysis. The methylation analysis was carried out Essay, as described previously (Jansson et al ., 1976; Chaplin, 1982; Ciucanu and Kerek, 1984). The permethylation was performed using sodium methylsulphinylmethanide and plants that in the methyl iodide. After treatment with sodium thiosulphate and extraction with chloroform, the permethylated material was hydrolysed with 2 M trifluoroacetic acid (2 h at 100°C). After concentration and reduction with sodium borodeuteride (1 h at 20°C), followed by Essay, acetylation with acetic anhydride (3 h at 120°C), the partially methylated alditol acetates were analysed by GLC (CP-Sil 43B column) with a temperature programme going from Races 140°C to 200°C at 4°C min −1 and by GLC-EIMS carried out on a MD800/8060 system (Fisons Instruments; 70 eV) equipped with a DB-1 fused silica capillary column (30 m × 0.32 mm; J and Essay W Scientific) at a temperature programme going from 140°C to 240°C at 4°C min −1 . Samples were exchanged twice in 99.9 atom% 2 H 2 O (Isotec) by successive lyophilization and finally dissolved in 99.96 atom% 2 H 2 O (Isotec). And Jamaica. All experiments were recorded on a three-channel Bruker AMX 500 MHz spectrometer (Bijvoet Center, Department of NMR Spectroscopy, Utrecht University, The Netherlands) equipped with an actively shielded triple-resonance ( 1 H 13 C 15 N) pulsed-field z-gradient probe at 67°C. The Holocaust Essay. 1 H NMR spectra were recorded with low-power presaturation at the water resonance for 1 s. Chemical shifts are expressed in ppm by and Oversized, reference to internal acetone (δ H 2.23) for 1 H and to the α-anomeric signal of external [ 13 C-1]-glucose (δ C-1 92.90) for 13 C. The following two-dimensional phase-sensitive experiments were recorded: TOCSYs (Braunschweiler and Ernst, 1983) with varying mixing times between 18 ms and 132 ms; NOESYs (Jeener et al ., 1979; Kumar et al ., 1980) with mixing times of Essay, 100 ms and 150 ms; and an undecoupled 1 H- 13 C HSQC (Bodenhausen and Ruben, 1980). A gradient-enhanced 1 H- 13 C HMBC (Bax and Summers, 1986) was also recorded. The following numbers of The Laws of the Essay, complex points were recorded (F 1 , F 2 ): 128 × 512 (TOCSY), 256 × 512 (NOESY), 128 × 512 (HSQC) and 128 × 1024 (HMBC), with averaging over The Holocaust Essay, 32 scans (TOCSY and NOESY), 256 scans (HSQC) and 400 scans (HMBC); spectral width (ω 1 , ω 2 ): 4000 Hz × 4000 Hz (TOCSY and NOESY), 16350 Hz × 4000 Hz ( 1 H- 13 C HSQC) and Millennium Development and Jamaica Essay 12577 Hz × 4000 Hz ( 1 H- 13 C HMBC) were used. A 60° shifted square sine-bell was used in all cases, with zero-filling once. All data were processed using triton software (Bijvoet Center, Utrecht University, The Netherlands). We are indebted to The Holocaust Essay, J. Lemoine for providing the chemical shifts of the EPS produced by S. thermophilus Sfi6. S.V. acknowledges stimulating discussions with S. Haseley. Blackwell Science Ltd, Oxford. Issue online: 1 March 2002 Version of Essay about, record online: 1 March 2002. 1 Ausubel, F.M. The Holocaust Essay. , Brent, R. , Kingston, R.E. , Moore, D.D. , Seidman, J.G. , Smith, J.A., et al ( 1994 ) Protocols in Molecular Biology . New York: John Wiley and Sons. 2 Bax, A. & Summers, M.F. A Worn Path Pdf. ( 1986 ) 1 H and 13 C assignments from The Holocaust sensitivity-enhanced detection of Digital, heteronuclear multiple-bond connectivity by The Holocaust, 2D multiple quantum NMR . Difference Primary. J Am Chem Soc 108 : 2093 – 2094 . CrossRef | CAS | Web of Science® 3 Betlach, M.R. , Capage, M.A. The Holocaust. , Doherty, D.H. , Hassler, R.A. , Henderson, N.M. The Laws Of The Races Essay. , Vanderslice, R.W., et al ( 1997 ) Genetically engineered polymers: manipulation of xanthan biosynthesis. In Industrial Polysaccharides. Genetic Engineering, Structure/Property Relations and Applications . Yalpani, M. (ed.). Amsterdam: Elsevier Science, pp. 35 – 50 . 4 Bock, K. The Holocaust. & Thøgersen, H. ( 1982 ) Nuclear magnetic resonance spectroscopy in the study of mono- and oligosaccharides . Annu Rep NMR Spect 13 : 1 – 57 . CrossRef 5 Bock, K. , Pedersen, C. , Pedersen, H. A Worn Path Pdf. ( 1983 ) Carbon-13 nuclear magnetic resonance data for oligosaccharides . Adv Carbohydr Chem Biochem 42 : 193 – 225 . CrossRef | Web of Science® 6 Bodenhausen, G. & Ruben, D.J. ( 1980 ) Natural abundance nitrogen-15 NMR by enhanced heteronuclear spectroscopy . Chem Phys Lett 69 : 185 – 189 . CrossRef | CAS | Web of Science® | ADS 7 Braunschweiler, L. & Ernst, R.R. ( 1983 ) Coherence transfer by isotopic mixing: application to proton correlation spectroscopy . The Holocaust Essay. J Magn Reson 53 : 521 – 528 . Web of Science® | ADS 8 Bubb, W.A. , Urashima, T. , Fujiwara, R. Millennium Development And Jamaica Essay. , Shinnai, T. , Ariga, H. ( 1997 ) Structural characterisation of the The Holocaust Essay exocellular polysaccharide produced by Streptococcus thermophilus OR 901 . Carbohydr Res 301 : 41 – 50 . CrossRef | PubMed | Web of vs. Traditional, Science® 9 Chaplin, M.F. ( 1982 ) A rapid and sensitive method for the analysis of Essay, carbohydrate components in glycoproteins using gas-liquid chromatography . A Worn Path Pdf. Anal Biochem 123 : 336 – 341 . CrossRef | PubMed | CAS | Web of Science® 10 Ciucanu, I. & Kerek, F. ( 1984 ) A simple and rapid method for the permethylation of carbohydrates . Carbohydr Res 131 : 209 – 217 . CrossRef | CAS | Web of The Holocaust, Science® 11 Ding, Y. , Kanie, O. , Labbe, J. , Palcic, M.M. , Ernst, B. , Hindsgaul, O. ( 1995 ) Synthesis and a worn path pdf biological activity of oligosaccharide libraries . Adv Exp Med Biol 376 : 261 – 269 . The Holocaust Essay. CrossRef | PubMed | CAS | Web of Science® Times Cited: 17 12 Doco, T. , Wieruszeski, J.-M. Primary. , Fournet, B. , Carcano, D. The Holocaust. , Ramos, P. , Loones, A. ( 1990 ) Structure of an extracellular polysaccharide produced by Streptococcus thermophilus . Carbohydr Res 198 : 313 – 321 . CrossRef | PubMed | Web of Science® 13 Dubois, M.A. , Gilles, K.A. , Hamilton, J.K. , Rebers, P.A. Plants That Dark. , Smith, F. The Holocaust Essay. ( 1956 ) Colorimetric method for determination of Races, sugars and related substances . Essay. Anal Chem 28 : 350 – 356 . Races. CrossRef | CAS | Web of Science® 14 Estrela, A.I. The Holocaust Essay. , De Pooley, H.M. , Lencastre, H. , Karamata, D. ( 1991 ) Genetic and biochemical characterization of Bacillus subtilis 168 mutants specifically blocked in the synthesis of the teichoic acid poly (3- O -beta-D-glucopyranosyl- N -acetylgalactosamine 1-phosphate): gneA , a new locus, is a worn path pdf associated with UDP- N -acetylglucosamine 4-epimerase activity . J Gen Microbiol 137 : 943 – 950 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 38 15 Faber, E.J. , Zoon, P. , Kamerling, J.P. , Vliegenthart, J.F.K. ( 1998 ) The exopolysaccharides produced by Streptococcus thermophilus Rs and The Holocaust Sts have the same repeating unit but differ in difference sources viscosity of their milk cultures . Carbohydr Res 310 : 296 – 276 . CrossRef | Web of Science® Times Cited: 69 16 Gerwig, G.J. , Kamerling, J.P. , Vliegenthart, J.F.G. ( 1978 ) Determination of the D and L configuration of neutral monosaccharides by high resolution capillary GLC . Carbohydr Res 62 : 349 – 357 . CrossRef | CAS | Web of Science® 17 Gerwig, G.J. Essay. , Kamerling, J.P. , Vliegenthart, J.F.G. Millennium Development Goals. ( 1979 ) Determination of the absolute configuration of monosaccharides in complex carbohydrates by capillary GLC . Carbohydr Res 77 : 1 – 7 . The Holocaust Essay. CrossRef | Web of Development and Jamaica, Science® 18 Gruter, M. The Holocaust. , Leeflang, B.R. , Kuiper, J. , Kamerling, J.P. , Vliegenthart, J.F.G. ( 1992 ) Structure of the exopolysaccharide produced by Lactococcus lactis subspecies cremoris H414 grown in a defined medium or skimmed milk . Young And Oversized. Carbohydr Res 231 : 273 – 291 . CrossRef | PubMed | Web of Science® 19 Gruter, M. , Leeflang, B.R. The Holocaust Essay. , Kuiper, J. , Kamerling, J.P. , Vliegenthart, J.F.G. ( 1993 ) Structural characterisation of the exopolysaccharide produced by Lactobacillus delbruckii subspecies bulgaricus rr grown in Digital vs. Traditional Art Essay skimmed milk . The Holocaust Essay. Carbohydr Res 239 : 209 – 226 . CrossRef | PubMed | CAS | Web of Science® 20 Jansson, P.-E. , Kenne, L. A Worn Path Pdf. , Liedgren, H. The Holocaust Essay. , Lindberg, B. , Lönngren, J. ( 1976 ) A practical guide to the methylation analysis of carbohydrates . That In The Dark. Chem Commun Univ Stockholm 8 : 1 – 74 . 21 Jeener, J. , Meier, B.H. Essay. , Bachmann, P. , Ernst, R.R. ( 1979 ) Investigation of exchange processes by two-dimensional NMR spectroscopy . That Grow. J Chem Phys 11 : 4546 – 4553 . Essay. CrossRef | Web of Science® Times Cited: 4087 | ADS 22 Kamerling, J.P. & Vliegenthart, J.F.G. A Worn Path Pdf. ( 1989 ) Carbohydrates. In Clinical Biochemistry — Principles, Methods, Applications, Vol. 1. Mass Spectrometry . Lawson, A.M. (ed.). The Holocaust Essay. Berlin: Walter de Gruyter, pp. 176 – 263 . Between Primary Sources. 23 Kolkman, M.A.B. The Holocaust. ( 1997 ) Capsular polysaccharide synthesis in Streptococcus pneumoniae . Thesis, Utrecht University, pp. 79 – 97 . 24 Kumar, A. , Ernst, R.R. , Wüthrich, K. ( 1980 ) A two-dimensional nuclear Overhauser enhancement (2D NOE) experiment for the elucidation of complete proton–proton cross-relaxation networks in biological macromolecules . Biochem Biophys Res Commun 95 : 1 – 6 . CrossRef | PubMed | CAS | Web of Science® 25 Lemoine, J. , Chirat, F. The Laws Races Essay. , Wieruszeski, J.-M. , Strecker, G. , Favre, N. , Neeser, J.-R. ( 1997 ) Structural characterization of the exocellular polysaccharides produced by Streptococcus thermophilus SFi39 and SFi12 . Appl Environ Microbiol 63 : 3512 – 3518 . PubMed | Web of Science® 26 Nagaoka, M. , Hashimoto, S. , Watanabe, T. , Yokokura, T. , Mori, Y. ( 1994 ) Anti-ulcer effects of lactic acid bacteria and their cell wall polysaccharides . The Holocaust. Biol Pharm Bull 17 : 1012 – 1017 . CrossRef | PubMed | CAS | Web of Science® 27 Nakajima, H. , Hirota, T. , Toba, T. , Itoh, T. , Adachi, S. ( 1992 ) Structure of the extra-cellular polysaccharide from slime-forming Lactococcus lactis subsp. cremoris SBT 0495 . Carbohydr Res 224 : 245 – 253 . CrossRef | PubMed | Web of Science® 28 Nakajima, H. , Toba, T. Of The. , Toyoda, S. ( 1995 ) Enhancement of antigen-specific antibody production by extracellular slime products from slime-forming Lactococcus lactis subspecies cremoris SBT 0495 in mice . Int J Food Microbiol 25 : 153 – 158 . CrossRef | PubMed | Web of Science® 29 Neeser, J.-R. & Schweizer, T.F. ( 1984 ) A quantitative determination by capillary gas-liquid chromatography of The Holocaust, neutral and The Laws of the Races Essay amino sugars (as O -methyloxime acetates), and a study on hydrolytic conditions for glycoproteins and polysaccharides in order to increase sugar recoveries . Anal Biochem 142 : 58 – 67 . CrossRef | PubMed | Web of Science® 30 Oda, M. , Nakamura, S. , Komatsu, S. Essay. , Kambe, M. , Tsuchiya, F. , Komiyama, K., et al ( 1982 ) Physiological activities of The Laws Essay, polysaccharide produced by Lactobacillus sp . The Holocaust. Jpn J Antibiot 35 : 2748 – 2754 . PubMed | CAS 31 Palcic, M.M. A Worn Path Pdf. & Hindsgaul, O. ( 1991 ) Flexibility in The Holocaust Essay the donor substrate specificity of β-1,4-galactosyltransferase: application in difference between primary the synthesis of complex carbohydrates . The Holocaust. Glycobiology 1 : 205 – 209 . CrossRef | PubMed | CAS 32 Roberts, I.S. ( 1995 ) Bacterial polysaccharides in that in the dark sickness and in health. The 1995 Fleming Lecture . Microbiology 141 : 2023 – 2031 . The Holocaust Essay. CrossRef | PubMed | Web of Science® Times Cited: 49 33 Robijn, G.W. , Thomas, J.R. , Haas, H. , Van Den Berg, D.J.C. , Kamerling, J.P. , Vliegenthart, J.F.G. ( 1995 a) The structure of the Goals exopolysaccharide produced by The Holocaust Essay, Lactobacillus helveticus 766 . Carbohydr Res 276 : 137 – 154 . CrossRef | PubMed | Web of Science® 34 Robijn, G.W. , Van Den Berg, D.J.C. , Haas, H. , Kamerling, J.P. Art Essay. , Vliegenthart, J.F.G. ( 1995 b) Determination of the The Holocaust structure of the exopolysaccharide produced by Essay about Young and Oversized, Lactobacillus sake 0-1 . Carbohydr Res 276 : 117 – 136 . CrossRef | PubMed | Web of Science® 35 Robijn, G.W. , Gutiérrez Gallego, R. , Van Den Berg, D.J.C. , Haas, H. The Holocaust Essay. , Kamerling, J.P. , Vliegenthart, J.F.G. ( 1996 a) Structural studies of the exopolysaccharide produced by Lactobacillus paracasei 34-1 . Carbohydr Res 285 : 129 – 139 . CrossRef | PubMed | Web of Science® 36 Robijn, G.W. And Jamaica. , Wienk, H.L.J. , Van Den Berg, D.J.C. , Haas, H. The Holocaust. , Kamerling, J.P. , Vliegenthart, J.F.G. ( 1996 b) Structural characterization of the sources exopolysaccharide produced by Essay, Lactobacillus acidophilus LMG9433 . Digital. Carbohydr Res 288 : 203 – 218 . CrossRef | PubMed | CAS | Web of Science® 37 Sandford, P.A. & Bard, J. ( 1983 ) Industrial utilization of polysaccharides. Essay. In The Polysaccharides , Vol. 2. Aspinall, G.O. A Worn Path Pdf. (ed.). New York: Academic Press, pp. The Holocaust Essay. 411 – 490 . Plants That. 38 Shaw, J.H. & Clewell, D.B. Essay. ( 1985 ) Complete nucleotide sequence of macrolide-lincosamide-streptogramin B-resistance transposon Tn 917 in Streptococcus faecalis . Millennium Development Goals And Jamaica. J Bacteriol 164 : 782 – 796 . PubMed | CAS | Web of Science® 39 Staaf, M. , Widmalm, G. , Yang, Z. The Holocaust Essay. , Huttunen, E. ( 1996 ) Structural elucidation of an Art Essay extracellular polysaccharide produced by Lactobacillus helveticus . Carbohydr Res 291 : 155 – 164 . CrossRef | PubMed | CAS 40 Stingele, F. & Mollet, B. ( 1996 ) Disruption of the gene encoding penicillin-binding protein 2b ( pbp2b ) causes altered cell morphology and cease in exopolysaccharide production in Streptococcus thermophilus Sfi6 . Mol Microbiol 22 : 357 – 366 . Wiley Online Library | PubMed | Web of Science® 41 Stingele, F. , Neeser, J.-R. , Mollet, B. ( 1996 ) Identification and characterization of the eps (exopolysaccharide) gene cluster from Streptococcus thermophilus Sfi6 . J Bacteriol 178 : 1680 – 1690 . PubMed | CAS | Web of Science® 42 Stingele, F. Essay. , Lemoine, J. , Neeser, J.-R. ( 1997 ) Lactobacillus helveticus Lh59 secretes an exopolysaccharide that is identical to the one produced by Lactobacillus helveticus TN-4, a presumed spontaneous mutant of Lactobacillus helveticus TY1-2 . Carbohydr Res 302 : 197 – 202 . CrossRef | PubMed | CAS | Web of Science® 43 Sutherland, I.W. ( 1998 ) Novel and established applications of microbial polysaccharides . Trends Biotechnol 16 : 41 – 46 . CrossRef | PubMed | CAS | Web of plants, Science® 44 Wlasichuk, K.B. , Kashem, M.A. , Nikrad, P.V. Essay. , Bird, P. , Jiang, C. , Venot, A.P. ( 1993 ) Determination of the specificities of rat liver Gal (β1–4) GlcNAc α-2,6-sialyltransferase and Gal (β1–3/4) GlcNAc α-2,3–sialyltransferase using synthetic modified acceptors . J Biol Chem 268 : 13971 – 13977 . PubMed | CAS | Web of primary and secondary, Science® 45 Yamamoto, Y. , Murosaki, S. , Yamauchi, R. , Kato, K. , Sone, Y. The Holocaust. ( 1994 ) Structural study on an exocellular polysaccharide produced by Lactobacillus helveticus TY1-2 . Carbohydr Res 261 : 67 – 78 . CrossRef | PubMed | Web of a worn path pdf, Science® 46 Yamamoto, Y. Essay. , Nunome, T. Essay About Young And Oversized. , Yamauchi, R. , Kato, K. The Holocaust Essay. , Sone, Y. The Laws Of The Essay. ( 1995 ) Structure of an exocellular polysaccharide of Lactobacillus helveticus TN-4, a spontaneous mutant strain of Lactobacillus helveticus TY1-2 . Essay. Carbohydr Res 275 : 319 – 332 . Millennium Development Goals And Jamaica Essay. CrossRef | PubMed | CAS | Web of Science® 47 Zopf, D. & Roth, S. The Holocaust Essay. ( 1996 ) Oligosaccharide anti-infective agents . Lancet 347 : 1017 – 1021 . 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Essay about The Holocaust - 879 Words | BartlebyAug 30 2011 Published by Cathy under Open University. The OU’s Advanced Creative Writing (A363) course ended in Essay, May. Or was it June? I can’t remember. Either way, I couldn’t wait for it to end. You may have mistaken the lack of any A363 entries on this blog being down to me being thoroughly engrossed in the course and The Laws Essay, spending every waking minute devouring the contents of the coursebook: A Creative Writing Handbook: Developing Dramatic Technique, Individual Style and Voice. The Holocaust Essay. This wasn’t the case. Essay About. I barely opened the book. It was drier and The Holocaust Essay, less engaging than the a worn path pdf poetry section of A215 (Creative Writing) and that’s not something to The Holocaust Essay be said lightly. It didn’t bode well from the start. After Googling my tutor I found out he was a professor in 19th Century literature. Hmm, not quite my kind of thing and, undoubtedly my thing wasn’t going to be his thing. Don’t get me wrong, my tutor was great. Difference Between Primary And Secondary Sources. He always gave timely feedback and was always there with a quick response if needed. At the day school he was encouraging and motivating. I love the day schools, I always come back full of The Holocaust, motivation and inspiration. Unfortunately, with A363, you only get two of them. I’d like far more, although I appreciate that some people only do courses with the OU because they don’t have to The Laws leave the house and Essay, see other people. The first TMA (a short story) came back with a mediocre mark. The next TMA was to be either a stage play, a radio play or a film/TV play based on and Oversized, the story we wrote for TMA1. This is where I look like a complete hypocrite and say that I absolutely loved the screenwriting bit of the book. The Holocaust. I got totally engrossed in Young and Oversized, learning how to write a stage play and The Holocaust Essay, decided that, although I had probably seen fewer than ten plays in my life, I was going to adapt my short story for the stage. I spent hours making notes, setting out where the actors would stand, what props I’d have, etc. Then I decided my story would be better as a radio play. I thoroughly read through the Digital vs. Traditional Art Essay pages in the coursebook on writing for The Holocaust the radio, borrowed a great book – Writing for Radio: A Practical Guide – from the Digital vs. Traditional Art Essay library, looked on the BBC Writersroom for scripts to read and template scripts to help with the layout, listened to radio plays on the radio and downloaded some from The Wireless Theatre Company and wrote my play. I received another mediocre mark. Ho hum. Essay. But I was well pleased with my play, so ner. TMA3 was a 1,000 word critique on The Laws of the Essay, another student’s work. Essay. This was the Millennium Goals and Jamaica TMA I was most dreading. I’m no good at critiques. My critiques begin with ‘I’ and end with either: Not quite the in-depth 1,000 word essay the OU were looking for. The Holocaust Essay. Still, I somehow managed to blag my way through it and primary and secondary sources, ended up with a good mark. For TMA4 we had to write a short (approx 500 words) proposal for our ECA, outlining briefly what it would be about. As the ECA wasn’t due in for months, I didn’t have a clue, as I’m more of a ‘I’ll decide the day before the assignment’s due what to The Holocaust Essay write’ kind of of the Races Essay, writer, not a ‘I’m going to meticulously plan this story months and months ahead’ one. But it was only a few hundred words, so on the day of the deadline, with a massive hangover, I typed out a few hundred words of rubbish. Essay. It wasn’t possible to a worn path pdf fail this TMA (well, it was, but only if you wrote fewer than 475 words, I think) but you did have to – at least loosely – stick with what you had proposed when it came to write the ECA. TMA5 was the second piece of original writing we had to do. Another short story. I’d actually had an The Holocaust idea of what I wanted to write for a while. I wanted to write about a goth dinner party where all the food was black. Yes, I know, not quite literary enough for primary and secondary sources the OU. Essay. My tutor didn’t like the slapstick ending (neither did I but that’s not the point), although he did say in the feedback that that was a personal thing. TMA6 was a 1,000 word extract of the Development ECA. I bashed out 1,000 words and received a deserved mediocre mark. By this time I was well and truly fed up with this course. I didn’t want to do it anymore. I’d found what looked like a great Creative and Professional Writing degree course at Canterbury Christ Church University and had applied for that. I was thrilled to be invited for an interview (they call it an audition, as it’s a performing arts college) and Essay, had hoped that after showing them my work (I’d taken a short story, my bagel poem, my radio play and a worn path pdf, a couple of pieces of flash fiction) that they would see what a genius I was and immediately offer me a place, despite me not even having one single CSE to my name and then I could pack in this stupid course. Unfortunately not. They gave me a conditional offer on the successful completion of A363. Bah. That meant I had to do the dreaded ECA. Essay. 4,000 words of Races Essay, a story I really didn’t want to write. I’d never written 4,000 words before. I like short, short stories. Ones under 1,000 words (although I think even 1,000 words is a lot to read in one go). I sort of cheated and wrote a 1,000 word each monologue for three characters, and rounding it off with the final 1,000 words being from the first character. I sent the ECA off and hoped they didn’t notice my story didn’t have the usual ‘beginning, middle and end, and a conflict to be resolved’ the OU are so fond of. They didn’t notice. The Holocaust Essay. Or, if they did, they weren’t too bothered. Millennium Development. I passed. The Holocaust. Not brilliantly, not terribly, but I passed. So, to about and Oversized sum up. The Holocaust. Did I enjoy the course? Apart from the The Laws Races Essay play writing bit, no, not at all. I couldn’t wait for it to be over. But plenty of The Holocaust Essay, people on my course did enjoy it. I only gave it the bare minimum of effort and my marks reflected this. I’d probably have enjoyed it more if I had worked harder, but I just didn’t engage with it. Now that the a worn path pdf OU’s behind me, I am massively looking forward to starting full-time education in a brick university, (I would say ‘real’ but I know how much that offends people who are with the OU) as well as being massively scared of it being three years’ long and being surrounded by 18 year olds. I’ll be blogging about Essay life as a ‘mature’ student here. Enjoyed this Cathy. Glad I wasn’t the only one to Millennium Development Goals and Jamaica Essay hate A363 and long for The Holocaust Essay it to be over. It has put me off the OU forever (in fact all distance learning). Communal moaning, groaning and celebrating etc on a daily face to face basis with fellow sufferers is jolly good therapy and very motivating. I am really envious of about, your place at The Holocaust Essay, Canterbury. I shall follow your blog and feedback with great interest. PS I bet you are not the oldest student on your course. Of The Essay. Lots of Essay, luck with it. Hi Cathy, enjoyed that a lot! I have to confess an interest here – I work in Marketing at difference between and secondary, Christ Church and helped to put together the ‘blurb’ for the Creative and Professional Writing Course. The Holocaust. But professional pride aside – I’m really looking forward to hearing about how you get on with it. And don’t worry, there are so many different types of people studying at UCF – you’re not in too much danger of Essay Young and Oversized, being surrounded by The Holocaust Essay 18 year olds! Myself, I’m starting the difference between and secondary sources part-time MA Creative Writing in Canterbury in about 3 weeks. Terrified. Starts with a short story module which is The Holocaust, so not my thing. Good tip about the short stories for i-phone – I will be downloading some this evening. Good luck anyway. Look forward to reading more of your stuff. L. […] White, a fellow student with whom I frequently correspond with on Twitter recently wrote her impressions of the course and she goes into much more detail than I have; it was she that spurred me on to write this piece. […] Only just seen your post, thought I must have a word too. I didn’t enjoy the Races course too much either, for different reasons though. My tutor didn’t seem to gel with my work at all, she left part way through the course, by which time, my most favourite TMA, a screen-play (that I had attempted both because the new format interested and The Holocaust Essay, stretched me). I received a bare-pass for Digital which more or less meant that I could only, at best secure a grade 2 pass…I didn’t feel the tutor’s comments were fair, and I wasn’t the only one in The Holocaust Essay, the tutor group who felt the same way. Goals And Jamaica Essay. Stuffing knocked out of me, I spent the rest of the course playing safe…I hate doing that, and felt I didn’t experiment or throw myself into Essay exploring style which is what I believe any course should encourage. I too passed the course with a grade 2. but I’m left feeling cheated as all the fun-stuff was torn off the bone by the lack of between primary, encouragement of the first tutor. Our replacement tutor was entirely different, it’s such a shame we didn’t have him from the The Holocaust start…so I sympathise with your views. So glad you passed, this year I am doing E301…it’s looking tough, but I’ll give it my best shot. I wish you all the very best, I’ve taken an interest in your writing from A215, A363 and feel you have enormous writing skills, which I’m sure you already know…good luck. best wishes, Liz. I think that the point of A363 was to make us think for ourselves about our own creativity. I galloped straight into the ECA miles b4 it needed to be in and then spent two months… ummm… reading books. Dark. cos I thought I orta do summat. I published my ECA in booklet form and hurled it around… it got good marks but no-one who read it liked it. The Holocaust Essay. Maybe cross-dressing dutcart-driving serial killers who fall in Digital Art Essay, love with sheep farmer serial killers aren’t to my pals’ tastes. hmmyah. The Holocaust. so I’m def thinking hard about my creativity, but am slowly filling up shopping trolleyloads of poetry books to return them to a worn path pdf the charity shops. Ezra Pound…. wot claptrap. Wordsworth…. we are supposed to understand these acclaimed creative geniusness but I don’t think I ever will. I’m studying law instead now. I’m glad you have decided to go full time to uni. The Holocaust. Good luck and Digital vs. Traditional Art Essay, keep in touch. If you ever renew your interest in radio drama, I heartily recommend William Ash’s The Way to Write Radio Drama. I loved A363, my tutor was excellent and, more importantly, gave me good marks. Essay. I think it was a course more about planning and editing though, rather than writing. Glad you are still writing though. And studying. I discovered your blog when I noticed on Webmaster Tools that you linked to mine. I couldn’t find the link, but was interested to see that you hated A363. Between. So did !! I got great marks for A215, but my A363 tutor and I obviously had different ideas. Essay. Like you, I really enjoyed the script writing. And the The Laws of the Races tip I really remember, and try to employ, is to vary my descriptions so they go from wide angle to zoom. (I go to a Creative Writing Adult Ed Class) Best wishes for Essay all you do. Do you still have a copy of the workbook? I’m starting in October and would love to get a head start! My email is above. Thanks! I enjoyed reading your experience on the A363 module. It was very well written, very honest, and vs. Traditional, did make me laugh out loud. (or should that be lol?) I’m on the course myself, I’ve had one assignment returned so far and The Holocaust Essay, it wasn’t a bad mark. But it’s early days so far, so I wont get too cocky! I’d love to read nmore of your stuff, how do I go about it? Flash Flood Journal. Funky Raw Magazine. Cook Vegetarian Magazine. Women's Running Magazine. Runner's World Magazine. Web Designer Magazine. Site Admin | Theme by Essay and Oversized Niyaz Cathy White Copyright © 2017 All Rights Reserved.