Clone and Expression of Mat Peptide Gene of 47 kD Pro tein in Orientia Tsutsugamushi Karp Strain
- VernacularTitle:恙虫病东方体Karp株相对分子质量为47蛋白基因的克隆与表达
- Author:
ping, CHANG
;
de-xin, ZHU
;
jiang, DU
;
zhi-chun, FENG
- Publication Type:Journal Article
- Keywords:
47 kD protein gene;
tsutsugamushi disease;
protein expression
- From:Journal of Applied Clinical Pediatrics
2004;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To know the expression of 47 kD protein of orientia tsutsugamushi(Ot) Karp strain and test its immunogenicity and antigenicity.Methods Forty-seven kD protein gene that code the whole mat peptide was amplified from the genomic DNA of Ot.Karp strain by PCR and constructed to a recombinant plasmid pGEX-47.The E. coli cells BL21 were transformed with pGEX-47,then the transformants were induced to express the recombinant 47 kD protein by IPTG. The antigenicity and immunogeni-(city) were identified by Dot Blot method and the experiment of immunizing mice respectively.Results 1. The PCR product of 47 kD protein gene of Ot.Karp strain was obtained and its sequence was the same as the published 47 kD protein gene.2. The E.coli expression plasmid pGEX-47 constructed was identified containing the 47 kD protein gene fragment by restrication analysis and PCR identification,and an expression band about 47 kD was detected by SDS-PAGE of the purified product from GST-fusson protein expressed by pGEX-47. 3.Dot Blot analysis and the experiment of immunizing mice testified its antigenicity and immunogenicity.Conclusions The 47 kD protein gene of Ot Karp strain is amplified that code the whole mat peptide, which can be expressed in prokaryotic cells and the expressed protein has antigenicity and immunogenicity.
