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- W2362819408 abstract "Nucleic acid vaccines are novel vaccination technologies emerged as 'Third Generation Vaccines'. Up to the present, most studies of the Helicobacter pylori (H.pylori) vaccines are focused on attenuated, killed or subunit vaccines, and the nucleic acid vaccine is seldom reported. Aims: To construct nucleic acid vaccine ofH.pyloriheat shock protein 60 (hsp60) gene, and to identify the immunogenicity of the protein expressed. Methods: The genomic DNA of standardH.pyloristrain CCUG17874 was extracted as the template, hsp60 gene was amplified by polymerase chain reaction (PCR) and cloned into pUCmT vector. After DNA sequence analysis, the amplified hsp60 gene was cloned into the eukaryotic expression vector pIRES through a series of restriction enzyme digestion and ligation reactions, then the recombinant plasmid was used to transform the competent E. coli DH5α, the positive clones were screened and identified by restriction enzyme digestion and PCR. The recombinant pIRES-hsp60 constructed was then transfected to eukaryotic cell COS-7 using LipofectamineTM 2000, the immunogenicity of HSP60 protein expressed by pIRES-hsp60 was detected by Western blotting. Results: Using the genomic DNA ofH.pylorias the template, 1 640 bp hsp60 gene fragment was amplified by PCR, sequence analysis showed that the coincidence rate was 98% as compared with the sequence ofH.pylorihsp60 in GenBank. It was confirmed by restriction enzyme digestion and PCR that hsp60 gene was cloned into the eukaryotic expression vector pIRES and the nucleic acid vaccine ofH.pylorihsp60 gene was successfully constructed. A specific stripe of HSP60 (60 kDa, 1 Da=0.992 1 u) expressed by pIRES-hsp60 was detected by Western blotting in transfected COS-7 cells. Conclusions: The nucleic acid vaccine ofH.pylorihsp60 gene is successfully constructed, its immunogenicity is also identified, which may pave the way for further investigating the immunologic effect of nucleic acid vaccine." @default.
- W2362819408 created "2016-06-24" @default.
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- W2362819408 date "2005-01-01" @default.
- W2362819408 modified "2023-09-27" @default.
- W2362819408 title "Construction of Nucleic Acid Vaccine of Helicobacter pylori Heat Shock Protein 60 Gene and Identification of its Immunogenicity" @default.
- W2362819408 hasPublicationYear "2005" @default.
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