Construction of Protokaryon Expression Vector Containing Tat and Xiap Genes
- VernacularTitle:pTAT-XIAP融合蛋白表达载体的构建
- Author:
Changwen JIANG
;
Chunbo XIA
;
Yajuan XU
- Publication Type:Journal Article
- Keywords:
TAT;
XIAP;
Transformation;
Fusion protein
- From:
Journal of Medical Research
2006;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the prokaryotic expression vector containing XIAP and TAT gene.Methods XIAP gene was synthesized and the pTAT-HA plasmid was digested with NcoI and XhoI restriction enzyme,then rat XIAP gene was ligated by T4 DNA ligase with pTAT-HA plasmid to construct pTAT-XIAP expression plasmid encoding XIAP and TAT fusion protein.pTAT-XIAP vector was transformed into E.coli.DH5?.Transformants were cultured on LB medium and selected.The plasmids were extracted,purified and identified by agarose gel electrophoresis.Results A few of transformants with pTAT-XIAP on LB medium containing ampicillin were observed and a number of them on LB medium without ampicillin,none of untrasformed DH5? colonies were found on LB medium with ampicillin.The gel electrophoresis analysis showed that the molecular weight of recombinated pTAT-XIAP vector was 4500bp and the pTAT-HA vector was about 3000bp consistent with the atlas of the vector.Conclusion XIAP gene was cloned into pTAT-HA vector,pTAT-XIAP vector was successfully constructed.
