中国中西医结合外科杂志2012，Vol.18Issue(3)：269-272,4.DOI:10.3969/j.issn.1007-6948.2012.03.015
固醇携带蛋白2腺病毒载体的构建与鉴定
Expression of Mouse SCP2 Gene Adenoviral Vector Carrying Albumin Promoter in Hepa1-6 Cells
摘要
Abstract
Objectives To construct the replication defective adenoviral vector of SCP2 gene carrying murine albumin promoter, and study the relations between SCP2 gene and the formation of cholesterol calculus. Methods The cDNA of SCP2 gene was cloned by using RT-PCR technique. The albumin promoter was linked to SCP2 gene's upstream, and the EGFP gene lied in its downstream. The plasmid pDC312-ALB-SCP2-IRES2 -EGFP was constructed by the gene recombination technique. The Admax Adenoviral Vector System was used to generate the replication defective adenoviral vectors, which were purified by CsCl method. The processes of TCID50 were applied to detect the titers of the adenoviral vectors. The RNA and protein were respectively extracted from the infected Hepal-6 cells by the adenoviral vector. The real-time quantitative PCR was employed to detect the mRNA expression levels, and the Western blotting analysis was used to measure the SCP2 protein levels. Result We constructed successfully the replication defective adenoviral vector of SCP2 gene carrying murine albumin promoter. When the mRNA levels of SCP2 gene were overexpressed, CYP7al mRNA levels were down-regulated (t=3.97,p<0.05); and the mRNA levels of HMGCR were up-regulated (t=3.23,p<0.05). Conclusions The SCP2 gene overexpression may affect cholesterol and bile acid metabolism, which could promote the formation of cholesterol calculus.关键词
SCP2基因/白蛋白启动子/腺病毒载体/胆固醇结石Key words
SCP2 gene/Albumin promoter/Adenoviral vector/Cholesterol calculus分类
生物科学引用本文复制引用
贾岩峰,崔云峰,崔乃强,彭雁飞,宁召臣,张琚..固醇携带蛋白2腺病毒载体的构建与鉴定[J].中国中西医结合外科杂志,2012,18(3):269-272,4.基金项目
天津市自然科学基金资助项目(08JCYBJC08700) （08JCYBJC08700）
