Prokaryotic expression of Helicobacter pylori gene hp1188
- VernacularTitle:幽门螺杆菌hp1188基因的原核表达
- Author:
Fei HAN
- Publication Type:Journal Article
- Keywords:
Helicobacter pylori;
hp1188 gene;
Expression
- From:
Journal of Chongqing Medical University
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To express the recombinant protein of the hp1188 gene of Helicobacter pylori NCTC 11637 in order to lay a foundation for the study on adhesion. Methods:The hp1188 gene was amplified from NCTC 11637 chromosomal DNA by PCR.The PCR product was inserted into the expression vector pQE-30.The recombinant vector pQE30-hp1188 was identified by DNA sequencing and transformed to E.coli DH5? for expression under IPTG induction.The recombinant protein was purified by Ni~(2+)-NTA agarose and identified by SDS-PAGE and western blot. Results:The sequence of hp1188 gene amplified was about 810 bp with a 98% sequence homology with hp1188 genes in GenBank. SDS-PAGE determined that the molecular weight of its expression products was 30 600 Dalton. The expression products of E.coli DH5? lyzate was about 50% of total soluble proteins,and the pellet also had expressed. Its purity was over 90% after purified with Ni~(2+)-NTA agarose and Western blot showed that the recombinant protein possesses fine binding capacity with the rabbit anti-H.pylori. Conclusion:hp1188 gene was successfully cloned and highly expressed in E.coli DH5?.