山东医药2011，Vol.51Issue(42)：17-18,2.
AQP5基因重组慢病毒载体的构建与鉴定
Construction and identification of recombinant lentiviral vector of AQP5 gene
摘要
Abstract
Objective To construct the recombinant lentiviral vector of AQP5 gene. Methods AQP5 gene sequence was amplified as target gene in vitro. The cDNA was inserted into plasmid pWPTS-GFP which was linearized by restriction endonucleases Mlu I and Sal I . The recombinant plasmid was transformed into competent DH5a celJs. The grown colonies were identified by colony PCR and DNA sequencing. Recombinant lentivector plasmids and the other helping plasmids were co-transfected into 293T cells by CaCl2, and cell culture supernatant was collected after 48 h. The virus supernatant was concentrated and titered in 293T cells. The infection efficiency of the constructed virus was determined in SGC790I cells. AQP5 expression in SGC7901 cells was determined by Western blot. Results DNA sequencing and Western blot demon strated that the lentivirus vector was constructed successfully. The constructed virus were obtained and infected SGC7901 cells. The titer of concentrated virus was 1×108 TU/ml. Conclusion Recombinant lentiu iral vector of AQP5 gene is suc cessfully constructed.关键词
水通道蛋白/SGC7901细胞/293T细胞/慢病毒载体Key words
aquaporins/ SGC7901 cells/ 293T cells/ lentiviral vector分类
医药卫生引用本文复制引用
张文杰,徐勇,王斌,徐泽宽,杨力,沈历宗,徐皓..AQP5基因重组慢病毒载体的构建与鉴定[J].山东医药,2011,51(42):17-18,2.基金项目
国家自然科学基金资助项目(30901421) （30901421）
江苏省卫生厅开放课题基金资助项目(XK03200903). （XK03200903）
