Bioinformatics analysis and prokaryotic expression of Strongyloides stercoralis serine protease inhibitor 1
10.16250/j.32.1374.2022285
- VernacularTitle:粪类圆线虫丝氨酸蛋白酶抑制剂1蛋白生物信息学 分析及原核表达
- Author:
Xue HAN
1
;
Xianglian BI
1
;
Hongying ZHAO
2
;
Yunliang SHI
1
;
Qing WEN
1
;
Jiayin LÜ
1
;
Jiayue SUN
1
;
Xiaoyin FU
1
;
Dengyu LIU
1
Author Information
1. Department of Parasitology, Guangxi Medical University, Key Laboratory of Basic Research on Regional Diseases in Guangxi Zhuang Autonomous Region, Nanning, Guangxi 530021, China
2. Department of Laboratory Medicine, Guangxi Zhuang Autonomous Region People’s Hospital, Nanning, Guangxi 530021, China
- Publication Type:Journal Article
- Keywords:
Strongyloides stercoralis;
Serine protease inhibitor 1;
Bioinformatics analysis;
Recombinant expression
- From:
Chinese Journal of Schistosomiasis Control
2023;35(3):244-250
- CountryChina
- Language:Chinese
-
Abstract:
Objective To predict the structure and antigenic epitope of the Strongyloides stercoralis serine protease inhibitor 1 (Ss-SRPN-1) protein using bioinformatics tools, and to construct prokaryotic expression plasmids for expression of recombinant Ss-SRPN-1 protein, so as to provide the basis for unraveling the function of the Ss-SRPN-1 protein. Methods The amino acid sequence of the Ss-SRPN-1 protein was downloaded from the NCBI database, and the physicochemical properties, structure and antigenic epitopes of the Ss-SRPN-1 protein were predicted using bioinformatics tools, including ExPASy, SWISS-MODEL and Protean. Primers were designed according to the nucleotide sequences of Ss-SRPN-1, and the Ss-SRPN-1 gene was amplified, cloned and sequenced with genomic DNA extracted from the infective third-stage larvae of S. stercoralis as a template. The Ss-SRPN-1 protein sequence was cloned into the pET28a (+) expression vector and transformed into Escherichia coli BL21 (DE) cells for induction of the recombinant Ss-SRPN-1 protein expression. The recombinant Ss-SRPN-1 protein was then purified and identified using Western blotting and mass spectrometry. Results Bioinformatics analysis showed that the Ss-SRPN-1 protein, which was composed of 372 amino acids and had a molecular formula of C1948H3046N488O575S16, was a stable hydrophilic protein, and the subcellular localization of the protein was predicted to be extracellular. The Ss-SRPN-1 protein was predicted to contain 11 dominant B-cell antigenic epitopes and 20 T-cell antigenic epitopes. The Ss-SRPN-1 gene with a length of 1 119 bp was successfully amplified, and the recombinant plasmid pET28a (+)/Ss-SRPN-1 was constructed and transformed into E. coli BL21(DE) cells. The expressed recombinant Ss-SRPN-1 protein had a molecular weight of approximately 43 kDa, and was characterized as a Ss-SRPN-1 protein. Conclusions The recombinant Ss-SRPN-1 protein has been expressed successfully, and this recombinant protein may be a potential vaccine candidate against strongyloidiasis.
