Reconstruction and prokaryotic expression of human high mobility group box 1 protein
- VernacularTitle:人高迁移率族蛋白的分子改建及原核表达
- Author:
Jing CHEN
;
Zhiqiang YUAN
;
Yizhi PENG
- Publication Type:Journal Article
- Keywords:
high mobility group box 1 protein;
clone;
mutation;
prokaryotic expression
- From:Journal of Third Military Medical University
2003;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To design and prepare recombinant mutant human high mobility group box 1 protein (HMGB1s) that can combine with HMGB1 receptors but cannot produce inflammatory effect, and accordingly lead to the creation of a new potential agent for anti-inflammatory therapy. Methods This experiment was based on successful clone and expression of human HMGB1.Six mutant HMGB1 cDNA were designed and constructed by one step inverse PCR. They were cloned into prokaryotic expressive vector pQE80L and followed with production of mutant HMGB1s and identification by Western blotting. Results Six mutant proteins were designed and constructed into prokaryotic expressive vector pQE80L. The recombinant HMGB1 proteins were obtained and identified by Western blotting. Conclusion Human HMGB1 mutants have been successfully constructed and the expression and characterization of intent proteins are identified. It will lay a foundation for further study on biological functions of HMGB1.
