Recombinant expression of multiepitope fusion polypeptide derived from Mycobacterium tuberculosis (Mtb.) antigens
- Author:
Chuan-Yong WU
1
Author Information
1. Department of Laboratory Diagnosis
- Publication Type:Journal Article
- Keywords:
Epitope;
Mycobacterium tuberculosis;
Synthetic;
Vaccines
- From:
Academic Journal of Second Military Medical University
2010;27(12):1281-1285
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct a recombinant multiepitope fusion polypeptide containing 6 HLA-A * 0201 restricted CTL epitopes derived from Mycobacterium tuberculosis (Mtb.) antigens and express it in E. coli. Methods: The full-length sequence encoding 6 HLA-A * 0201 restricted CTL epitopes were derived from Mtb. antigens Rv03O9, Rv0173, and Ag85A, Ag85C; pan DR Th epitope (PADRE), Trojan peptide, and furin-sensitive linker RVKR were synthesized, amplified by polymerase chain reaction(PCR), inserted into the expression vector pGEX-4T-1, and transformed into E. coli BL21 (DE3). After induced by isopropylthio-β-D-galactoside (IPTG), the recombinant protein expression was examined by SDS-PAGE and Western blot. Results: The recombinant multiepitope fusion polypeptide plasmid was successfully constructed and expressed(about 40 000) in E. coli BL21 (DE3). Conclusion: The recombinant multiepitope fusion polypeptide may provide a basis for developing novel TB vaccine.
