Construction and verification of eukaryotic expression vector for mouse Hsp84
- VernacularTitle:小鼠Hsp84真核表达质粒的构建及鉴定
- Author:
Yewei JI
- Publication Type:Journal Article
- Keywords:
Hsp90;
Eukaryotic expression;
Stress endurance
- From:
Journal of Chongqing Medical University
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct the eukaryotic expression vector of mouse Hsp84.Methods:Amplify the Hsp84 gene by reverese transcription and touchdown PCR,and then the PCR products was ligated into pMD18-T vector.The positive was selected and verified by agorose electrophoresis and sequencing.The positive clone was used as template and further amplified with the primers containing endodigestion sites and protective bases in both terminals.The PCR products were then inserted into eukariotic expression vector pcDNA3.1.DH-5? was transformed with ligated plasmids.Check for the presence of positive clones by means of agorose electrophoresis and endodigestion.Results:The sequence inserted into the recombined positive T vectors was corresponding to the BALb/c mouse Hsp84 gene provided in Gene bank.The presence of positive eukaryotic expression vector was verified by double endodigestion.Conclusions:The eukaryotic expression plasmid for mouse Hsp84 was successfully constructed.
