Construction and identification of eukaryotic expressed plasmid vector pLenti6/V5- DEST-Cx43 in human
- VernacularTitle:人真核表达的质粒载体pLenti6/V5-DEST-Cx43的构建及鉴定
- Author:
Huimin FAN
;
Yang LI
;
Zhongmin LIU
- Publication Type:Journal Article
- Keywords:
Connexin 43 Lentivirus Eukaryotic cells Gene expression
- From:
Chinese Journal of Thoracic and Cardiovascular Surgery
2003;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct eukaryotic expressed plasmid vector pLenti6/V5- DEST-Cx43 in human for further study on bioinformative communication between myocardial cell, embryonic development of heart and differentiation of myocardial cell. Methods Cx43 cDNA fragments were obtained from plasmid p18-T by restriction enzyme BamH1、Xho1, after the agarose gel electrophoresis was performed, the Cx43 cDNA was retrieved. pENTR11 were extracted by enzyme BamH1、Xho1, then connected with Cx43 cDNA by T4 DNA ligase. LR reaction was performed and Cx43 cDNA were cloned into Lentiviral vector pLenti6/V5-DEST. Results Agarose electrophoresis and sequent examination, identified that Cx43 cDNA was cloned into Lentiviral vector pLenti6/V5-DEST. Conclusion The present experiment demonstrate a successful cloning of human Cx43 cDNA into the Lentiviral vector pLenti6/V5-DEST.