Construction and identification of expression of superantigen SED mutants
- VernacularTitle:超抗原SED突变体的构建表达和鉴定
- Author:
Yafei LI
;
Xihua ZHU
;
Yunhui HUANG
;
Jin YANG
;
- Publication Type:Journal Article
- Keywords:
superantigen;
staphylococcal enterotoxin D;
mutants
- From:Journal of Third Military Medical University
1984;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct staphylococcal enterotoxin D (SED) mutants expressed in Escherichia coli ( E. coli ). Methods The expected mutants were introduced into the SED DNA by megaprimer PCR method. The PCR products ligated to plasmid pTrcHis B were transformed into E. coli DH5? for IPTG induced expression. The target protein was purified by Ni NTA metal affinity chromatography and analyzed by SDS PAGE and immunoblotting. Results The sequencing results showed that mutant nucleic acids were successfully introduced at the expected sites of SED gene. SDS PAGE and immunoblotting confirmed that the proteins of SED mutants were obtained by Ni NTA metal affinity chromatography. The mutants were named as SEDN23A, SEDN23A/H26R, SEDF45A, SEDL59A, SEDN61A, SEDI92A, and SEDF203A, respectively. Conclusion Several SED mutants are successfully constructed, which lays a foundation for subsequent studies of immune recognition of SED.
