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891.
Vesicle-Mediated Transfer of Virulence Genes from Escherichia coli O157:H7 to Other Enteric Bacteria 总被引:4,自引:0,他引:4 下载免费PDF全文
Membrane vesicles are released from the surfaces of many gram-negative bacteria during growth. Vesicles consist of proteins, lipopolysaccharide, phospholipids, RNA, and DNA. Results of the present study demonstrate that membrane vesicles isolated from the food-borne pathogen Escherichia coli O157:H7 facilitate the transfer of genes, which are then expressed by recipient Salmonella enterica serovar Enteritidis or E. coli JM109. Electron micrographs of purified DNA from E. coli O157:H7 vesicles showed large rosette-like structures, linear DNA fragments, and small open-circle plasmids. PCR analysis of vesicle DNA demonstrated the presence of specific genes from host and recombinant plasmids (hly, L7095, mobA, and gfp), chromosomal DNA (uidA and eaeA), and phage DNA (stx1 and stx2). The results of PCR and the Vero cell assay demonstrate that genetic material, including virulence genes, is transferred to recipient bacteria and subsequently expressed. The cytotoxicity of the transformed enteric bacteria was sixfold higher than that of the parent isolate (E. coli JM109). Utilization of the nonhost plasmid (pGFP) permitted the evaluation of transformation efficiency (ca. 103 transformants μg of DNA−1) and demonstrated that vesicles can deliver antibiotic resistance. Transformed E. coli JM109 cells were resistant to ampicillin and fluoresced a brilliant green. The role vesicles play in genetic exchange between different species in the environment or host has yet to be defined. 相似文献
892.
Grey mullet, Chelon labrosus , (60–100 g) have been found to respond immunologically to Cryptocotyle lingua , following a single exposure to 20000 cercariae, by the production of humoral antibody, sensitized pronephric leucocytes and cytotoxic serum factors. Antibody titres measured by passive haemagglutination reached a peak at week 4 with a -log2 titre of 16±S.E. 1.0, and titres of 10.7±S.E. 1.0 were still recorded after 10 weeks at the termination of the experiment. Cercarial agglutination was found unreliable as a rapid test. Pronephric leucocytes, sensitive to cercarial antigen when measured by the under-agarose migration method, were detected between weeks 1 and 6, peaking at week 2. In vitro polarization was increased when cells were incubated with the antigen, but this increase was not significantly different between control and infected fish. Heat-labile cytotoxic factors of immune sera have been demonstrated to whole cercariae in vitro , these factors being associated with structural damage to the tegument. Pronephros cells isolated from immune fish during each week of infection showed no evidence of adherence to cercariae or metacercariae in vitro. The results are discussed in relation to both host response and course of infection of the parasite. 相似文献
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Isoforms of soybean seed oil body membrane protein 24 kDa oleosin are encoded by closely related cDNAs 总被引:3,自引:0,他引:3
Andrzej Kalinski Deborah S. Loer Jane M. Weisemann Benjamin F. Matthews Eliot M. Herman 《Plant molecular biology》1991,17(5):1095-1098
We have characterized two cDNA clones for 24 kDa soybean oleosin, the seed oil body membrane protein. Differences in the predicted amino acid sequences of the two clones and the presence of a doublet on immunoblots indicate that 24 kDa oleosin exists in at least two isoforms in soybean. The predicted amino acid sequence also contains a unique carboxy terminal region that is dominated by a series of different tandem amino acid repeats. 相似文献
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Chrissa A. Dwyer Toshihiko Katoh Michael Tiemeyer Russell T. Matthews 《The Journal of biological chemistry》2015,290(16):10256-10273
Protein O-mannosylation is a glycan modification that is required for normal nervous system development and function. Mutations in genes involved in protein O-mannosyl glycosylation give rise to a group of neurodevelopmental disorders known as congenital muscular dystrophies (CMDs) with associated CNS abnormalities. Our previous work demonstrated that receptor protein-tyrosine phosphatase ζ (RPTPζ)/phosphacan is hypoglycosylated in a mouse model of one of these CMDs, known as muscle-eye-brain disease, a disorder that is caused by loss of an enzyme (protein O-mannose β-1,2-N-acetylglucosaminyltransferase 1) that modifies O-mannosyl glycans. In addition, monoclonal antibodies Cat-315 and 3F8 were demonstrated to detect O-mannosyl glycan modifications on RPTPζ/phosphacan. Here, we show that O-mannosyl glycan epitopes recognized by these antibodies define biochemically distinct glycoforms of RPTPζ/phosphacan and that these glycoforms differentially decorate the surface of distinct populations of neural cells. To provide a further structural basis for immunochemically based glycoform differences, we characterized the O-linked glycan heterogeneity of RPTPζ/phosphacan in the early postnatal mouse brain by multidimensional mass spectrometry. Structural characterization of the O-linked glycans released from purified RPTPζ/phosphacan demonstrated that this protein is a significant substrate for protein O-mannosylation and led to the identification of several novel O-mannose-linked glycan structures, including sulfo-N-acetyllactosamine containing modifications. Taken together, our results suggest that specific glycan modifications may tailor the function of this protein to the unique needs of specific cells. Furthermore, their absence in CMDs suggests that hypoglycosylation of RPTPζ/phosphacan may have different functional consequences in neurons and glia. 相似文献
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