Expression of fused P-glycoprotein in E. coli DH5?
- VernacularTitle:P-糖蛋白的融合表达研究
- Author:
Xiangwei WANG
;
Tangjun YANG
;
Lixin ZHANG
;
Chunhai LI
;
- Publication Type:Journal Article
- Keywords:
multidrug resistance;
P glycoprotein;
expression
- From:Journal of Third Military Medical University
1984;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the expression of the extracellular fragment of P glycoprotein in the bacteria. Methods According to the P glycoprotein antigen analyses and mdr1 gene structure, a couple of primers were designed to amplify the extracellular DNA fragment of the protein with PCR. The product was inserted into pGEM T vector followed by sequencing. The sequencing verified vectors were cloned into pGEX 2T to get a highly expression recombinant vector pGEX Pgp. The recombinant was transferred into E. coli DH5?, and its expression, identification and purification were studied. Results The highest level of recombinant expression was achieved at 4 h after IPTG induction. SDS PAGE analysis indicated that the expressed protein was about 30?10 3. Conclusion The target gene fragment expression at high level is established for the preparation of its antibody and biospanning of its specific peptide by using random phage library, founding a basis for the targeting treatment of multidrug resistance.
