王磊
用基因组学和功能基因组学包括生物信息学的手段进行微生物进化,细菌脂多糖功能基因和致病菌的快速鉴定方面
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- 姓名:王磊
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博士生导师
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学科领域:
微生物学
- 研究兴趣:用基因组学和功能基因组学包括生物信息学的手段进行微生物进化,细菌脂多糖功能基因和致病菌的快速鉴定方面
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王磊, Lu Fenga, b, Jiang Taoa, Hongjie Guoa, Jianguo Xuc, Yayue Lia, Fedousi Rezwand, Peter Reevesd, Lei Wanga, *
Microbial Pathogenesis 36(2004)109-115,-0001,():
-1年11月30日
Shigella strains are human pathogens. The O antigen gene cluster of Shigella dysenteriae O7 was sequenced and analyzed. It contains genes for synthesis of nucleotide sugars including UDP-2-acetamido-2-deoxy-D-galacturonamide, UDP-2-acetamido-2-deoxy-Dgalacturonic acid and dTDP-4-amino-4,6-dideoxy-D-glucose. Also found in the gene cluster are genes encoding O unit flippase, O antigen polymerase and sugar transferases. The Escherichia coli O121 O antigen, which is present in an important Shiga toxin-producing strain, has the same structure as that of S. dysenteriae O7, and we found that the gene clusters also had the same genes and organization. Four genes specific to S. dysenteriae O7 and E. coli O121 were identified by PCR screening against representatives of 186 E. coli (including Shigella) O serotypes. E. coli O121 and S. dysenteriae O7 isolates can be distinguished by PCR of the H antigen fliC gene.
Molecular typing, Shigella dysenteriae O7
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【期刊论文】Structural and Genetic Characterization of the Shigella boydii Type 13 O Antigen
王磊, Lu Feng, , †, Sof'ya N. Senchenkova, Jinghua Yang, Alexander S. Shashkov, Jiang Tao, Hongjie Guo, Guang Zhao, Yuriy A. Knirel, Peter Reeves, and Lei Wang, *
JOURNAL OF BACTERIOLOGY, Jan. 2004, p. 383-392,-0001,():
-1年11月30日
er membrane of gramnegative bacteria and plays an important role in pathogenicity. The chemical structure and genetic organization of the S. boydii type 13 O antigen were investigated. The O polysaccharide was found to be acid labile owing to the presence of a glycosyl phosphate linkage in the main chain. The structure of the linear pentasaccharide phosphate repeating unit (O unit) was established by nuclear magnetic resonance spectroscopy, including two-dimensional COSY, TOCSY, ROESY, and H-detected 1H,13C and 1H,31P HMQC experiments, along with chemical methods. The O antigen gene cluster of S. boydii type 13 was located and sequenced. Genes for synthesis of UDP–2-acetamido-2,6-dideoxy-L-glucose and genes that encode putative sugar transferases, O unit flippase, and O antigen polymerase were identified. Seven genes were found to be specific to S. boydii type 13. The S. boydii type 13 O antigen gene cluster has higher levels of sequence similarity with Vibrio cholerae gene clusters and may be evolutionarily related to these gene clusters.
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王磊, LEI WANG, HEATHER CURD, WENJIA QU, AND PETER R. REEVES*
JOURNAL OF CLINICAL MICROBIOLOGY, Nov. 1998, p. 3182-3187,-0001,():
-1年11月30日
Shiga toxin (Stx)-producing Escherichia coli strains of serogroup O111 are the most frequently isolated non-O157 strains causing outbreaks of gastroenteritis with hemolytic-uremic syndrome. The O111 O-antigen gene cluster had been cloned and about half of it has been sequenced; we have now sequenced the remainder of the gene cluster, which is 12.5 kb in length and which comprises 11 genes. On the basis of sequence similarity, we have identified all the O-antigen genes expected, including five sugar biosynthetic pathway genes, three transferase genes, the O-unit flippase gene, and the O-antigen polymerase gene. By PCR testing with E. coli strains representing all 166 O-antigen forms, some randomly selected gram-negative bacteria, and Salmonella enterica serovar Adelaide, we showed that four O-antigen genes are highly specific to O111. This work provides the basis for a sensitive test for the rapid detection of E. coli O111. This is important both for decisions related to patient care, because early treatment may reduce the risk of life-threatening complications, and for the detection of sources of contamination.
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王磊, LEI WANG, AND PETER R. REEVES*
INFECTION AND IMMUNITY, Aug. 1998, p. 3545-3551,-0001,():
-1年11月30日
The O157:H7 clone of Escherichia coli, which causes major, often prolonged outbreaks of gastroenteritis with hemolytic-uremic syndrome (HUS) such as those in Japan, Scotland, and the United States recently, is thought to be resident normally in cattle or other domestic animals. This clone is of major significance for public health and the food industry. We have developed a fast method for sequencing a given O antigen gene cluster and applied it to O157. The O157 O antigen gene cluster is 14 kb in length, comprising 12 genes and a remnant Hrepeat unit. Based on sequence similarity, we have identified all the necessary O antigen genes, including five sugar biosynthetic pathway genes, four transferase genes, the O unit flippase gene, and the O antigen polymerase gene. By PCR testing against all 166 E. coli O serogroups and a range of gram-negative bacterial strains, including some that cross-react serologically with E. coli O157 antisera, we have found that certain O antigen genes are highly specific to O157 E. coli. This work provides the basis for a sensitive test for rapid detection of O157 E. coli. This is important both for decisions on patient care, since early treatment may reduce the risk of life-threatening complications, and for detection of sources of contamination. The method for fast sequencing of O antigen gene clusters plus an ability to predict which genes will be O antigen specific will enable PCR tests to be developed as needed for other clones of E. coli or, once flanking genes are identified, clones of any gram-negative bacterium.
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王磊, Lei Wang, †, Sandy Huskic, Adam Cisterne, Deborah Rothemund, and Peter R. Reeves*
JOURNAL OF BACTERIOLOGY, May 2002, p. 2620-2625,-0001,():
-1年11月30日
Escherichia coli O55 is an important antigen which is often associated with enteropathogenic E. coli clones. We sequenced the genes responsible for its synthesis and identified genes for O-antigen polymerase, O-antigen flippase, four enzymes involved in GDP-colitose synthesis, and three glycosyltransferases, all by comparison with known genes. Upstream of the normal O-antigen region there is a gne gene, which encodes a UDP-GlcNAc epimerase for converting UDP-GlcNAc to UDP-GalNAc and is essential for O55 antigen synthesis. The O55 gne product has only 20 and 26% identity to the gne genes of Pseudomonas aeruginosa and E. coli O113, respectively. We also found evidence for the O55 gene cluster's having evolved from another gene cluster by gain and loss of genes. Only three of the GDP-colitose pathway genes are in the usual location, the other two being separated, although nearby. It is thought that the E. coli O157:H7 clone evolved from the O55:H7 clone in part by transfer of the O157 gene cluster into an O55 lineage. Comparison of genes flanking the O-antigen gene clusters of the O55:H7 and O157:H7 clones revealed one recombination site within the galF gene and located the other between the hisG and amn genes. Genes outside the recombination sites are 99.6 to 100% identical in the two clones, while most genes thought to have transferred with the O157 gene cluster are 95 to 98% identical.
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王磊, Lei Wang, †, Kanella Andrianopoulos, , Dan Liu, Michel Y. Popoff, and Peter R. Reeves*
JOURNAL OF BACTERIOLOGY, Mar. 2002, p. 1669-1677,-0001,():
-1年11月30日
The 46 serogroups of Salmonella enterica have different O-antigens, and each is thought to have a specific form of the O-antigen cluster. Comparison of the 145 serovars of serogroup B revealed much more intraserogroup genetic diversity than expected. The O27 factor, due to an 1-6 linkage between O units in place of the more common α 1-2 linkage and previously thought to be due to a converting bacteriophage, is now shown to be due to a wzyα(1-6) gene located within the major gene cluster. Surprisingly a remnant of this gene in all O27- serovars shows that the ancestor was O27+. There are six distinct gene cluster forms, five apparently derived by a series of deletions and one by an insertion from an ancestral O27+ form present in 57 serovars. The history of the gene cluster and movement between subspecies I and II can be traced. Two of the derivative forms still have a functional wzyα(1-6) gene, while in three it has been inactivated by deletion or insertion. Two of the forms lacking a functional wzyα(1-6) gene have the wzyα(1-2) gene first described for strain LT2 as rfc, whereas for the third the wzy gene has not been located.
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王磊, LEI WANG, WANG, WENJIA QU, AND PETER R, REEVES*
INFECITON AND IMMUNITY, NOV. 2001, p. 6923-6930,-0001,():
-1年11月30日
Shigella straine are in realityclones of Escherichia coli and are belived to have emerged relatively recently (G. M. Pupo, R. Lan, an P. R. Reeves, Proc, Natl, Acad. Sci, USA 97: 10567-10572, 2000).There are 33 O-antigen forms in these shigella clones, of which 12 are identical O antigens of other E. coli strains. We sequenced O-antigen gene cluters from Shigella boydii serotypes 4, 5, 6, and 9and also studied the O53-and O79-antigen gene clusters of E. coli, encoding O antigens identical to those of S. boydii serotype 4 and S. boydii serotype 5, respectively, In both cases the, S. boydii and E. coli O-antigen gene clusters have the same genes and organization, The clusters of both S, Boydii 6 and S. boydii 9 O antigens have atypical features, with a functional insertion sequence and a wzx gene located in the orientation opposite to that of all other genes in S, boydii Serotype 9 and an rmiC gene located away from other rml genes in S. boydii sdrotye 6. Sequences of O-antigen gene clusters from another three shigella clones have been published, and two of them also have abnormal structures, with either the entire cluster or one gene being located on a plasmid in Shigella sommei or Shigella dysemteriae, respectively, It appears that a high proportion of clusters coding for o antigens spectifc to Shigella clones have atypical features, perhaps indicating recent formationof thene clusters.
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【期刊论文】Sequence of E. Coli O104 antigen gene cluser and identification of O104 specific genes
王磊, Lei Wang a, , Connie E. Briggs b, Deborah Rothemund a, Pina Fratamico B, John B. Huchansky B, Peter R. Reeves a, *
L Wang et al./Gene 270(2001)231-236,-0001,():
-1年11月30日
The Escherichia coli O104 polysacchride is an important antigen, which contains sialic acid and is often assccited with EHEC clones, Sialic acid is a coponent of many animal tissues, and its presence in hacterial polysaccharides may comtribute to bacerial pathogenicity, We sequenced the genes responsible for O104 antigen synthesis and have found genes which from their seuences are idenified as an O antigen polymerase gene, an O antigen flippase gene, three CMP-sialic acid synthesis genes, and three potential glycosyl trans ferase genes, The E. coli K9 group IB capsular antigen bas thesame structure as the O104 O antigen, and we find using gene by gene PCR that the K9 gene chuster is essentially the same s that for O104. It appears that the distinction between presence as gtroup IB capsule or O antigen for this sturchure dous not involve any difference in gees present in the O antigen gene chrster, By PCR testiong against representative strains for the 166 E, coli O antigens and some randomly selected Gram-negative bacteria, we identified three O antigen genes which are highly specifkc to O104/K9. This work provids the Basis for a sensitive test for rapid detection of O104 E. coli, This is important both for decisions on patient care as carly treatment may reduce the risk of life-threatcning complications and for a faster response in comtrol of food borne outhreaks.
Polymerase chain reaction (, PCR), , Pathogeic E., coli: Molecular typing: Serotyping
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王磊, LEI WANG AND PETER R, REEVES*
JOURNAL OF CLINICAL MIROBIOLOGY, Sept, 2000, p, 5256-5261,-0001,():
-1年11月30日
O antigen is part of the lipopoly saccharide present in the outer membrane of gram-megative bacteria, Escherichia coli and salmonella enteeerica each have many forms of O antigen, hut only three are common to the tow species, It has been found that, in general, O-antigen genes are of low GC content, This deviation in GC content from that of typical S, enterica or E, coli genes (51%) is thought to indicate that the O-antigen DNA originated in species other than S, ENTERIC OR E. coli and was captured by lateral transfer, The O-antigen structure of Salmonella enterica O35 is identical to that of coli O111, commonly found in enteropathogenic E, coli strains, This O, antigen, which has been shown to be a virulence factor in E. coli, contains colitose, a 3. 6-dideoxyhexose found only rarely in the Enteroacteriaceae, Sequencing of the O35-antigen cluster of S. enterica serovar Adelaide revealed the same gene order and flanking genes as in E. coli O111 The divergence between corresponding gense of these two gene clusters at the nucleotide level ranges from 21.8 to 11.7%, within the normal range of divergence between S. enterical and E. coli, We conclude that the ancestor of E. coli and S. enterica had O antigen identical to the O111 and O35 antigens, respectively, of these species and that the gene cluster encoding it has survived in both species.
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王磊, LEI WANG, DEBORAH ROTHEMUND, HEATHER CURD, AND PETER R. REEVES*
JOURNAL OF CLINICAL MIROBIOLOGY, May 2000, p, 1786-1790,-0001,():
-1年11月30日
Flagellar (H) antigens are mostly encoded by genes at the fliC locus in E, coli, We have sequenced 11 H7 fliC genes from Escherichia coli strains that belong to seven O serotypes, These sequences, together with those of nine other H7 fliC genes (from strains of three different O serotypes) drwirmvrf trvrmy;u (S.D. Reid, R. K. Selander, and T.S. Whittam, J, Bacteriol, 181:153-160, 1999), include 10 different sequences, The differencs betweem thses 10 sequences range from 0.06 to 3.12%. By comparison with other E. coli flagellin genes, we have identified primer length sequences specific for H7 gense in gensral and others specific for H7 genes of O157 and O55 stains: the specificity was confirmed by PCR testing the type strains for all 53 E, coli H types, We have Previously identified genes specific for the E. coli O157 antigen, and use of the combination of O157-and H7-specific primers allows the sensitive and rapid detecition of O157:H7 E. colo strains, which cause the majority of hemorrhagic clitos cases.
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