%0 Journal Article
%T A Novel System for Producing Lentiviral Vectors<br>一种新型慢病毒载体制备方法的建立
%A MA Qiang
%A LI Ming
%A DONG Wen-Qi
%A WU Ying-Song
%A <br>马强
%A 李明
%A 董文其
%A 吴英松
%J 生物工程学报
%D 2007
%I 
%X The aim was to develop a cell culture system capable of producing high titer lentiviral vector stocks with recombinant vaccinia viruses as helpers. BHK21 was co-transfected by three main plasmids containing the transducing plasmid pVECRNA, the packaging plasmid pGAGPOL and the envelop plasmid pVSVG, and thereafter infected with the vaccinia vTF-3 containing bacteriophage T7 RNA polymerase gene using Lipofectamine2000. After 4 days incubation, the culture supernatant of lentiviral vectors was collected and judged by RT-PCR and the Western blot, the results showed that lentiviral vectors were found in the culture supernatant; approximately (11.71 +/- 0.80) x 10(11) copies of lentiviral vector RNA were present per mL of cell culture supernatant, as detected by Real-time PCR; the vector stocks with titers was up to (1.3 +/- 0.18) x 10(8) tu/mL, as detected by flow cytometry , which is one order of magnitude higher than the output of classical manufacture system. These results suggest that the new poxviral/lentiviral hybrid system for efficient lentiviral vector production was initially established. It provides the basis for the future development of industrial application.
%K lentiviral vector
%K vaccinia virus
%K gene therapy<br>慢病毒载体
%K 痘苗病毒
%K 基因治疗
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=41D15872C73C1B31&yid=A732AF04DDA03BB3&vid=EA389574707BDED3&iid=94C357A881DFC066&sid=808D6B9EB5A8B4B4&eid=40700C9CB4E84E3B&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=5