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| 1 | Screening Helicobacter pylori genes induced during infection of mouse stomachs显示文摘AIM:To investigate the effect of in vivo environment on gene expression in Helicobacter pylori(H.pylori) as it relates to its survival in the host.METHODS:In vivo expression technology(IVET) systems are used to identify microbial virulence genes.We 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 pIVET12.Our novel IVET systems were developed by the fusion of random Sau3A DNA fragments of H.pylori and a tandem-reporter system of chloramphenicol acetyltransferase and beta-galactosidase.Additionally,each vector contains a kanamycin resistance gene.We used a mouse macrophage cell line,RAW 264.7 and mice,as selective media to identify specific genes that H.pylori expresses in vivo.Gene expression studies were conducted by infecting RAW 264.7 cells with H.pylori.This was followed by real time polymerase chain reaction(PCR) analysis to determine the relative expression levels of in vivo induced genes.RESULTS:In this study,we have identified 31 in vivo induced(ivi) genes in the initial screens.These 31 genes belong to several functional gene families,including several well-known virulence factors that are expressed by the bacterium in infected mouse stomachs.Virulence factors,vacA and cagA,were found in this screen and are known to play important roles in H.pylori infection,colonization and pathogenesis.Their detection validates the efficacy of these screening systems.Some of the identified ivi genes have already been implicated to play an important role in the pathogenesis of H.pylori and other bacterial pathogens such as Escherichia coli and Vibrio cholerae.Transcription profiles of allivi genes were confirmed by real time PCR analysis of H.pylori RNA isolated from H.pylori infected RAW 264.7 macrophages.We compared the expression profile of H.pylori and RAW 264.7 coculture with that of H.pylori only.Some genes such as cag A,vac A,lpx C,mur I,tlp C,trx B,sod B,tnp B,pgi,rbf A and inf B showed a 2-20 fold upregulation.Statistically significant upregulation was obtained for all the above mentioned genes(P < 0.05).tlp C,cag A,vac A,sod B,rbf A,inf B,tnp B,lpx C and mur I were also significantly upregulated(P < 0.01).These data suggest a strong correlation between results obtained in vitro in the macrophage cell line and in the intact animal.CONCLUSION:The positive identification of these genes demonstrates that our IVET systems are powerful tools for studying H.pylori gene expression in the host environment. | 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 | 2012 | World Journal of Gastroenterology2012，18，32： | 5 |
| 2 | Numerical taxonomic study of fast-growing soybean rhizobia and proposal that Rhizobium fredii be assigned to Sinorhizobium gen nov显示文摘 | CHEN W X YAN G H LI J L | 1988 | Int J Syst Bacteriol1988，38，： | 3 |
| 3 | Protective effects of free radical inhibitors in intracerebral hemorrhage in rat显示文摘 | Peeling J Yan H J Chen S G | 1998 | Brain Res1998，795，12： | 2 |
| 4 | Direct determination of fourteen underivatized amino acids from Whitmania pigra by using liquid chromatography-evaporative light scattering detection显示文摘 | Yan D Li G Xiao X H | 2007 | Journal of Chromatography A2007，1138，： | 1 |
| 5 | Shear stress induces endothelial differentiation from a murine embryonic mesenchymal progenitor cell line显示文摘 | WANG H RIHA G M YAN S | 2005 | Arteriosclerosis Thrombosis and Vascular Biology2005，25，9： | 1 |
| 6 | Genetic polymorphism of Malus sieversii populations in Xinjiang, China 显示文摘 | Yan G R Long H Song W Q | 2008 | Genet Resour Crop Evol2008，，55： | 1 |
| 7 | Comparison of efficacy of selamectin, ivermectin and mebendazole for the control of gastrointestinal nematodes in rhesus macaques, China显示文摘 | WANG T YANG G Y YAN H J | 2008 | Vet Parasitol2008，153，12： | 1 |
| 8 | Determination of near coincident site lattice orientation in MgO/Cu composite显示文摘 | Wang Y G Zhang Z Yan G H | 2002 | Journal of Materials Science2002，37，： | 1 |
| 9 | Thermodynamics of interaction between cationic gemini surfactants and hydrophobically modified polymers in aqueous solutions 显示文摘 | Bai G Wang Y Yan H | 2002 | JPhys Chem B2002，106，9： | 1 |
| 10 | The effect of electric field, annealing temperature and filler loading on the percolation threshold of polystyrene containing carbon nanotubes and graphene nanosheets 显示文摘 | Pang H Chen C Zhang Y C Ren P G Yan D X Li Z M | 2011 | Carbon2011，49，6： | 1 |
| 11 | Effects of acute aerobic exercise on multiple aspects of executive function in preadolescent children 显示文摘 | CHEN A G YAN J YIN H C | 2014 | Psychology of Sport and Exercise2014，15，6： | 1 |
| 12 | Influence of droplet kinetic energy flux density from fixed spray-plate sprinklers on soil infiltration,runoff and sediment yield显示文摘 | Yan H J Bai G He J Q | 2011 | Biosystems Engineering2011，110，2： | 1 |
| 13 | Chemical analysis of Astragalus mongholicus polysaccharides and antiox- idant activity of the polysaccharides 显示文摘 | YAN H XIE Y P SUN S G | 2010 | Carbohydrate Polymers2010，82，3： | 1 |
| 14 | Enhancement of the light output power of InGaN/GaN lightemitting diodes grown on pyramidal patterned sapphire substrates in the micro-and nanoscale显示文摘 | Gao H Y Yan F W Zhang Y Li J M Zeng Y P Wang G H | | 0，，01： | 1 |
| 15 | Effects of Yb3+on the corrosion resistance and deposition rate of electroless Ni-P deposits显示文摘 | Yan M Ying H G Ma T Y | 2008 | Applied Surface Science2008，255，5： | 1 |
| 16 | The performances of BMI nanocomposites filled with nanometer SIC显示文摘 | Yan H X Ning R C Liang G Z Ma X Y | 2005 | Journal of Applied Polymer Science2005，95，5： | 1 |
| 17 | A comprehensive e valuation method of manufacturing process resources and environmental attributes and its application显示文摘 | YAN W ZHANG H JIANG Z G | 2009 | Key Engineering Materials2009，，： | 1 |
| 18 | Cytotoxicity of carbon nanomaterials:single-wall nanotube,multi-wall nanotube,and fullerene显示文摘 | Jia G Wang H Yan L | 2005 | Environ Sci Technol2005，39，5： | 1 |
| 19 | Effects of heat treatment on mechanical properties of H13 steel显示文摘 | Yan G H Huang X M Wang Y Q | 2010 | Metal Science and Heat Treatment2010，52，7： | 1 |
| 20 | Sin显示文摘 | Yan X Z Kuang X Y Mao A J Kuang F G Wang Z H Sheng X W 2013 Acta Phys | 2013 | 62 107402 (in Chinese)2013，，： | 1 |
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