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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 | CXCR4 regulates growth of both prima ry and metastatic breast cancer显示文摘 | Smith M C Luker K E Garbow J R Prior J L Jackson E Piwnica-Worms D | 2004 | Cancer Res2004，64，： | 1 |
| 3 | A multicenter, open, non-comparative phase Ⅱ study of a combination of fludarabine phosphate,cytarabine and granulocyte colonystimulating factor in relapsed and refractory acute myeloid leukemia and de novo refractory anemia with excess of blasts in transformation 显示文摘 | Jackson G Taylor P Smith GM | 2001 | Br J Haematol2001，112，1： | 1 |
| 4 | Percutaneous subclaviancentral venous catheterization in children younger than oneyear of age显示文摘 | Finck C Smith S Jackson R | 2002 | Am Surg2002，68，4： | 1 |
| 5 | A muhicenter,open, non- comparative phase lI study of a combination of fludarabine phosphate, cytarabine and granulocyte colony-stimulating factor in relapsed and refractory acute myeloid leukemia and de novo refractory anaemia with excess of blasts in transformation 显示文摘 | Jackson G Taylor P Smith GM | 2001 | Br J Haematol2001，112，1： | 1 |
| 6 | TheLunar Orbiter Laser Altimeter Investigation on theLunar Reconnaissance Orbiter Mission 显示文摘 | Smith D E Zuber M T Jackson G B | 2010 | SpaceScience Reviezv2010，150，： | 1 |
| 7 | Electrical alternans and vulnerability to ventricular arrhythmias显示文摘 | JACKSON LE SMITH JM | 1994 | N Engl J Med1994，330，4： | 1 |
| 8 | Adjunctive treatment of eon- genital pigmented nevi with phenol chemical peel 显示文摘 | Hopkins JD Smith AW Jackson IT | 2000 | Plast Recon- str Surg2000，105，1： | 1 |
| 9 | A multicentre, open, non-comparative phase Ⅱ study of a combination of fludarabine phosphate, cytarabine and granulocyte colony-stimulating factor in relapsed and refractory acute myeloid leukemia and de novo refractory anaemia with excess of blasts in transformation 显示文摘 | Jackson G Taylor P Smith GM | 2001 | Br J Haematol2001，112，1： | 1 |
| 10 | Competitive and noncompetitive inhibition of the DNA-dependent protein kinase显示文摘 | Izzard RA Jackson SP Smith GC | | 0，，： | 1 |
| 11 | Multiple-locus variable number tandem repeat analysis reveals genetic relationships within Bacillus anthracis 显示文摘 | Keim P Price LB Klevytska AM Smith KL Schupp JM Okinaka R Jackson PJ Hugh-Jones ME | 2000 | J Bacteriol2000，182，10： | 1 |
| 12 | The association of sex with outcomes among patients undergoing primary percutaneous coronary intervention for ST elevation myocardial infarction in the contemporary era:Insights from the Blue Cross Blue Shield of Michigan Cardiovascular Consortium(BMC2)显示文摘 | Jackson EA Moscucci M Smith DE | 2011 | Am Heart J2011，161，： | 1 |
| 13 | Neutralization of muscarinic receptor autoantibodies by intravenous immunoglobulin in Sjogren zyndrome 显示文摘 | Smith A J Jackson MW Wang F | 2005 | Hum Immunol2005，66，4： | 1 |
| 14 | Understanding the Microdynam- its of Farm Structural Change: Entry, Exit, and Re- structuring Among Wisconsin Family Farmers in the 1980S显示文摘 | Jackson - Smith D B | 1999 | Rural Sociology1999，64，1： | 1 |
| 15 | Effects of Inequality and Reasons for Inequality on Group Identification and Cooperation in Social Dilemmas显示文摘 | Smith E R Jackson J W Sparks C W | | 0，，02： | 1 |
| 16 | The association of sex with outcomes among patients undergoing primary per- cutaneous coronary intervention for ST elevation myocardial infarction in the contemporary era: Insights from the Blue Cross Blue Shield of Michigan Cardiovascular Consortium (BMC2)显示文摘 | Jackson EA Moscucci M Smith DE | 2011 | Am Heart J2011，161，1： | 1 |
| 17 | Electrical altemans andvulnerability to ventricular arrhythmias 显示文摘 | Rosenbaum DS Jackson LE Smith JM | 1994 | NEnglJMed1994，330，： | 1 |
| 18 | Nutrient Losses by Surface Runoff Following the Application of Organic Manures to Arable Land Nitrogen显示文摘 | Smith K A Jackson D R Pepper T J | 2001 | Environment Pollution2001，112，： | 1 |
| 19 | Principles for national and regional guidelines on cardiovascular disease prevention: a scientific statement from the World Heart and Stroke Forum显示文摘 | Smith SC Jr Jackson R Pearson TA | 2004 | Circulation2004，109，25： | 1 |
| 20 | Microstrip patch designs thatdo not excite surface waves 显示文摘 | Jackson D R Williams J T Bhattacharyya A K Smith RL Buchheit S J Long S A | 1993 | IEEE Trans on Anten-nas and Propagation1993，41，8： | 1 |
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