Construction of mice angiopoietin-1 gene lentiviral expression vector by Gateway technology and its virus packa-ging
10.3969/j.issn.1000-3606.2013.09.017
- VernacularTitle:Gateway技术构建小鼠血管生成素-1慢病毒表达载体及其病毒包装
- Author:
Qiuping LI
;
Xinna MA
;
Xiaoying ZHANG
;
Jing XU
;
Shen ZHANG
;
Chunzhi WNAG
;
Zhichun FENG
- Publication Type:Journal Article
- Keywords:
angiopoietin-1;
lentiviral;
vector;
Gateway technology
- From:
Journal of Clinical Pediatrics
2013;(9):866-870
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a lentiviral vector carrying angiopoietin-1 (Ang-1) and DsRed gene, and to package a virus particles. Methods The Ang-1 lentiviral vector with DsRed (PLV.Ex3d.P/puro-CMV>Ang-1>IRES/DsRed-Express2) was constructed by Gateway technology, and identiifed by PCR and gene sequencing. The lentiviral vector was mixed with helper vector pLV/helper-SL3, pLV/helper-SL4 and pLV/helper-SL5 by Lipofectamine 2000 to prepare DNA-Lipofectamine?2000 complexes. The complexes were then added to transfect 293FT cells and package virus. The virus titers and infection ef-ifciency were determined by lfuorescence expression. Results Ang-1 lentiviral vector PLV.Ex3d.P/puro-CMV>Ang-1>IRES/DsRed-Express2 was constructed successfully as identified by PCR and gene sequencing. Lentivirus with high-efficiency infection was produced by transfection to 293FT cells and the virus titer was 5×108 TU/ml. Conclusions The recombinant len-tiviral vector for Ang-1 was successfully constructed by Gateway technology and the lentivirus with high-efifciency infection packaging can be used for further experiment of Ang-1 gene.
