基础医学与临床2011，Vol.31Issue(5)：485-489,5.
PNPLA3基因表达以及干扰腺病毒的构建及鉴定
The adenovirus construction and identification of expression and interference of PNPLA3 gene
摘要
Abstract
Objective To construct a adenovirus vector of PNPL43 expression and interference, a gene related to metabolism of glucose and lipid and liver fatty disease.Methods The expression and interferce primers were designed according to PNPLA3 gene sequence, the sequences were cloned into pAdTrack-CMV or pAdTrack-U6 vector with Bgl Ⅱ and Xho Ⅰ restriction site.E.coliBJ5183 sensitive bacterias were cotransfected with lined vector cutted by Pine Ⅰ enzyme and adenovirus vector pAdEasy-l.The obtained recombinant adenovirus vector was cutted with Pac Ⅰ enzyme, then adenovirus was obtained in 293A cells transfected with lined recombinant adenovims plasmaids.The titer of virus was measured based on the expression level of green fluorescent protein.The transfection efficiency of green fluorescent protein into mouse primary hepatocytes was calculated.Results DNA sequencing and digestion identification demonstrated that the expression and interference adenovirns vectors of PNPLA3 gene were constructed.The titer of concentrated virus was 1.5 × 1011VP/mL.The infection efficiency reached above 90％ and interference efficiency decreased endogenous PNPLA 3 mRNA in hepatocytes by more than 80％ when mouse primary hepatocytes were stabled transfected with 120 infection multiplicity.Conclusion The expression and interference adenovims vector of PNPL43 gene is successfully constructed.关键词
PNPLA3/过表达/干扰/腺病毒Key words
PNPL43( patatin-like phospholipase domain-containing 3 )/ overexpression/ interference/ aclenovirus分类
生物科学引用本文复制引用
乔爱君,方福德,常永生..PNPLA3基因表达以及干扰腺病毒的构建及鉴定[J].基础医学与临床,2011,31(5):485-489,5.基金项目
国家重点基础研究发展计划项目(973计划)(2011CB504004) （973计划）
国家自然科学基金(30971079,30800389,90919019) （30971079,30800389,90919019）
