EXPRESSION AND IDENTIFICATION OF HEAT SHOCK PROTEIN 70 IN E.COLI
- VernacularTitle:人源性热休克蛋白70在大肠杆菌中的表达及鉴定
- Author:
Xiaoxuan NING
;
Kaichun WU
;
Yua LI
- Publication Type:Journal Article
- Keywords:
heat shock protein 70;
clone;
gene expression
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
To express a gene of heat shock protein 70(HSP70) in E.coli , HSP70 gene was amplified by PCR. The PCR products were cloned into pUCm T vector and were sequenced; The HSP70 gene was subcloned into vector pET 21a(+) and expressed in E.coli. SDS PAGE and Western blot were employed to identify the expression of the HSP70 gene. Results showed that a fragment about 1 96kb was amplified by PCR. Sequence analysis revealed that the sequence of HSP70 was correct. The HSP70 gene was cloned into pET 21a(+) identified by enzyme digestion and PCR. SDS PAGE and Western blot showed that a M 72 000 protein was expressed and could be recognized by anti HSP70 antibody. Therefore,HSP70 gene has been successfully expressed in E.coli .
