Objective To construct the human interleukin 18 DNA plasmids vaccine and to express the eukaryotic plasmids vaccine in mammalian cell lines Cos 7 and D5.Methods Gene recombinant technique was used to construct hIL 18 ...Objective To construct the human interleukin 18 DNA plasmids vaccine and to express the eukaryotic plasmids vaccine in mammalian cell lines Cos 7 and D5.Methods Gene recombinant technique was used to construct hIL 18 eukaryotic expression vectors.Calcium phosphate method was performed to transect recombinant hIL 18 eukaryotic expression vectors into Cos 7 and D5 cells. In situ hybridization and Western Blot were implemented to verify the transient expression of recombinant hIL 18 in Cos 7 and D5.Results The eukaryotic expression plasmid pVAX1 IL 18 was constructed successfully.hIL 18 was transiently expressed in Cos 7 and D5.Conclusion The eukaryotic expression plasmid pVAX1 IL 18 was constructed. In situ hybridization and Western Blot results proved the successful transient expression of pVAX1 IL 18 in Cos 7 and D5.Therefore,the work has settled the foundation for further biological research on hIL 18,including immunogene therapy through hIL 18.展开更多
The cDNA of human Interleukin\|18 was obtained by RT\|PCR from the mRNA of human peripheral blood cells.The DNA sequencing was performed and the DNA was cloned into expression vector pBV220.The overexpression was perf...The cDNA of human Interleukin\|18 was obtained by RT\|PCR from the mRNA of human peripheral blood cells.The DNA sequencing was performed and the DNA was cloned into expression vector pBV220.The overexpression was performed in E.coli HB101.The expressive product was purified through Sephadex G100 and identified by N\|terminal amino acid sequencing and Western blot.展开更多
文摘Objective To construct the human interleukin 18 DNA plasmids vaccine and to express the eukaryotic plasmids vaccine in mammalian cell lines Cos 7 and D5.Methods Gene recombinant technique was used to construct hIL 18 eukaryotic expression vectors.Calcium phosphate method was performed to transect recombinant hIL 18 eukaryotic expression vectors into Cos 7 and D5 cells. In situ hybridization and Western Blot were implemented to verify the transient expression of recombinant hIL 18 in Cos 7 and D5.Results The eukaryotic expression plasmid pVAX1 IL 18 was constructed successfully.hIL 18 was transiently expressed in Cos 7 and D5.Conclusion The eukaryotic expression plasmid pVAX1 IL 18 was constructed. In situ hybridization and Western Blot results proved the successful transient expression of pVAX1 IL 18 in Cos 7 and D5.Therefore,the work has settled the foundation for further biological research on hIL 18,including immunogene therapy through hIL 18.
文摘The cDNA of human Interleukin\|18 was obtained by RT\|PCR from the mRNA of human peripheral blood cells.The DNA sequencing was performed and the DNA was cloned into expression vector pBV220.The overexpression was performed in E.coli HB101.The expressive product was purified through Sephadex G100 and identified by N\|terminal amino acid sequencing and Western blot.
