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Screening helicobacter pylori genes induced during infection of mouse stomachs

World Journal of Gastroenterology 2012年第32期18卷 4323-4334页

作者： Aparna Singh Nathaniel Hodgson Ming Yan Jungsoo Joo Lei Gu Hong Sang Emmalena Gregory-Bryson William G Wood Yisheng Ni Kimberly Smith Sharon H Jackson William G Coleman Laboratory of Biochemistry and Genetics National Institute of Diabetes and Digestive and Kidney Diseases National Institutes of Health Bethesda MD 20892 United States Laboratory of Host Defenses National Institute of Allergy and Infectious Diseases National Institutes of Health Bethesda MD 20892 United States

AIM:To investigate the effect of in vivo environment on gene expression in helicobacter pylori(***) as it relates to its survival in the ***:In vivo expression technology(IVET) systems are used to identify microbial virulence *** modified the IVET-transcriptional fusion vector,pIVET8,which uses antibiotic resistance as the basis for selection of candidate genes in host tissues to develop two unique IVET-promoter-screening vectors,pIVET11 and *** novel IVET systems were developed by the fusion of random Sau3A DNA fragments of *** and a tandem-reporter system of chloramphenicol acetyltransferase and ***,each vector contains a kanamycin resistance *** used a mouse macrophage cell line,RAW 264.7 and mice,as selective media to identify specific genes that *** expresses in *** expression studies were conducted by infecting RAW 264.7 cells with *** was followed by real time polymerase chain reaction(PCR) analysis to determine the relative expression levels of in vivo induced ***:In this study,we have identified 31 in vivo induced(ivi) genes in the initial *** 31 genes belong to several functional gene families,including several well-known virulence factors that are expressed by the bacterium in infected mouse *** factors,vacA and cagA,were found in this screen and are known to play important roles in *** infection,colonization and *** detection validates the efficacy of these screening *** of the identified ivi genes have already been implicated to play an important role in the pathogenesis of *** and other bacterial pathogens such as Escherichia coli and Vibrio *** profiles of allivi genes were confirmed by real time PCR analysis of *** RNA isolated from *** infected RAW 264.7 *** compared the expression profile of *** and RAW 264.7 coculture with that of *** *** ge

关键词： helicobacter pylori in vivo expression technology virulence genes mice infection

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