Construction of an eukaryotic expression vector encoding human granzyme B and it's expression in Hep2 cells
- VernacularTitle:人颗粒酶B真核表达载体的构建与表达分析
- Author:
Xiuying LI
;
Liangping XIA
;
Jinwei XIE
;
Suqing ZHAO
;
Zhongyuan ZHENG
;
Haitao ZHANG
;
Qiongmei JI
;
Minyou LI
;
Zheny ZHU
- Publication Type:Journal Article
- Keywords:
Granzyme B;
Lymphocytes;
Hep2 cells
- From:
Chinese Journal of Pathophysiology
2000;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct pVAX1-GrB. METHODS: Lymphocytes from human laryngeal carcinoma tissue were separated from tumor tissue. The fragment of granzyme B (GrB) was amplified by RT-PCR and was recombined to the downstream of T7 promoter in the vector pVAX1. The construction was transfected into Hep2 cells with lipofectamine 2000. The expression of protein was identified by indirect immunofluorescent antibody assay. RESULTS: It has been proved that the sequence of the RT-PCR product was totally consistent with the data of GenBank by DNA sequencing analysis. The GrB cDNA fragment was cloned into the vector of pVAX1 in the right direction and the open reading fragment of GrB was maintained. The target protein was detected in the transfected Hep2 cells. CONCLUSION: The pVAX1-GrB plasmid was successfully constructed and expressed. [
