Construction of Tre enzyme eukaryotic expression vector and identification of its function in loxLTR sequence-specific recognition
- VernacularTitle:Tre 酶真核表达载体构建及其特异性识别 loxLTR序列的功能鉴定
- Author:
Qingquan LIU
;
Zhuoran YU
;
Yongtao SUN
- Publication Type:Journal Article
- Keywords:
Tre enzyme;
Eukaryotic expression vector;
Human immunodeficiency virus-1;
LoxLTR sequence;
Electroporation
- From:
Journal of Medical Postgraduates
2014;(12):1254-1257
- CountryChina
- Language:Chinese
-
Abstract:
Objective Clearing HIV provirus is the key to cure AIDS .The study was to construct the Tre enzyme eukaryotic expression vector and identify its function in specific recognition of loxLTR sequence in HIV provirus . Methods Tre gene was in-serted into eukaryotic expression vector pcDNA 3.1 gene recombination manipulation by genetic recombination techniques including gene synthesis , PCR, restriction enzyme digestion and ligation .EGFPpA-LoxLTR sequence was inserted into pmCherry-N1 vector and was tested by restriction enzyme digestion , PCR and sequencing .Constructed vectors were electroporated into HeLa cells , then using fluorescence microscopy to observe fluorescence intensity changes . Results PCR, restriction enzyme digestion , electrophoresis and sequencing confirmed that Tre enzyme eukaryotic expression vector had been constructed successfully , and it could specifically recog-nize and cut loxLTR sequence after being transfected into Hela cells . Conclusion Constructed Tre enzyme eukaryotic expression vector can be expressed in Hela cells and specifically recognize loxLTR sequence , which has prepared the experimental ground for fur-ther studies of clearing HIV provirus .
