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- W2370086804 abstract "Objective To construct a prokaryotic expression system of Salmonella paratyphi A H1a gene, and to determine the immunogenicity and immunoprotection of the expressed product rH1a and the frequencies of H1a gene carrying and expressing in S .paratyphi A isolates. Methods The H1a gene of a clinical S. paratyphi A strain JH01 was amplified using high fidelity PCR. The cloned H1a gene was sequenced after T-A cloning. A prokaryotic expression system of the H1a gene, pET32a-H1a-E coli BL21DE3, was then constructed. SDS-PAGE and BioRad Agarose Image Pattern Analysis System were applied to examine the rHla expression. Ni-NTA affinity chromatography was performed to collect rH1a. The immunogenicity of rH1a was determined by Western blot. PCR and FLISA were established to detect the carrying and expressing frequencies of the H1a genes in 98 S. paratyphi A isolates. The immunoprotective effects of rH1a in S. paratyphi A strain 50001 infected mice were observed. Results In comparison with the reported corresponding sequences, both the nucleotide and putative amino acid sequence homologies of the cloned H1a gene were 99.59% . The expression output of rH1a was approximately 60% of the total bacterial proteins. S. paratyphi A antiserum could recognize as well as combine with rH1a. rH1a could efficiently induce rabbits to produce specific antibody. 100%(98/98) of the S.paratyphi A isolates had H1a gene and expressed H1a. Half of the mice that oral-taken or subcutaneous injected 500μg with rH1a for immunization were survival after challenged S. paratyphi A strain 50001. And when co-immunized with 5μg rLTB, the survival rates of the mice increased to 75.0% and 66.7%, respectively. Conclusion A prokary- otic expression system with high efficiency of H1a gene from a clinical S. paratyphi strain was successfully constructed in this study. rH1a showed a fine immunogenicity and a certain immunoprotective effect. H1a is an extensively distributed and frequently expressed gene in different clinical S. paratyphi strains." @default.
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- W2370086804 date "2006-01-01" @default.
- W2370086804 modified "2023-09-26" @default.
- W2370086804 title "Construction of prokaryotic expression system of Salmonella paratyphi A H1a gene and the immunogenicity and inanunoprotection of the expressed product" @default.
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