中文摘要：

目的将人水甘油通道蛋白9（AQP9）真核表达质粒pEGFP-N1-AQP9及干扰质粒pshRNA-AQP9通过电穿孔转化减毒沙门菌株SL7207,并验证其表达情况,为阐明AQP9在非酒精性脂肪肝病（NAFLD）中的作用奠定基础。方法从人肝脏组织中通过RT-PCR获得AQP9基因,克隆到载体pEGFP-N1上,构建真核表达质粒pEGFP-N1-AQP9,设计合成针对人AQP9的siRNA,退火形成双链shRNA,并插入pGenesil-1质粒中,构建干扰质粒pshRNA-AQP9,电穿孔转化减毒沙门菌株SL7207,通过PCR、Western blot验证其表达情况,并检测其稳定性。结果成功构建pEGFP-N1-AQP9及pshRNA-AQP9重组质粒,将其转化SL7207后,能在重组沙门菌中表达。结论成功构建SL7207/pEGFP-N1-AQP9及SL7207/pshRNA-AQP9重组沙门菌,为进一步研究AQP9对NAFLD的基因治疗奠定基础。

英文摘要：

Objective To transform human aquaglyceroporin 9（AQP9） recombinant eukaryotic expression plasmid pEGFP-N1-AQP9 and interference plasmid pshRNA-AQP9 into attenuated Salmonella strain SL7207 by electroporation and detect their expression,in order to lay the foundation for elucidating the human AQP9 in pathogenesis of nonalcoholic fatty liver disease（NAFLD）.Methods AQP9 gene was obtained by RT-PCR from human hepatic tissue and cloned into pEGFP-N1 vector,recombinant eukaryotic expression plasmid pEGFP-N1-AQP9 was constructed,designed and synthesized human AQP9 SiRNA,annealed to double-stranded ShRNA,inserted to pGenesil-1 plasmid,interference plasmid pshRNA-AQP9 was constructed,and then transformed into attenuated Salmonella strain SL7207 by electroporation,the expression were detected by PCR and Western Blot,and we detected their stability.Results Recombinant plasmid pEGFP-N1-AQP9 and pshRNA-AQP9 were constructed,transformed into attenuated Salmonella strain SL7207,after transformed into SL7207,could express in recombinant Salmonella.Conclusion Recombinant Salmonella SL7207/pEGFP-N1-AQP9 and SL7207/pshRNA-AQP9 are successfully constructed,lay the foundation for further study on the gene therapy of NAFLD with AQP9.