To obtain the protein of duck interferon alpha and study its biological activities, the prokaryotic expression vector of DuIFN-αwas constructed and expressed in BL21 (DE3) plysS. Using PCR technique, the protein gene...To obtain the protein of duck interferon alpha and study its biological activities, the prokaryotic expression vector of DuIFN-αwas constructed and expressed in BL21 (DE3) plysS. Using PCR technique, the protein gene of DuIFN-αwas cloned from pMD-18-duIFN-αrecombinant. The gene was then inserted to pGEM-T vector and identified by restriction endonuclease analysis and sequencing. DuIFN-αwas ligated with the prokaryotic expression vector of pET30 a, then transformed into BL21 (DE3) plysS. The best inducing time and IPTG concentration for the expression of this recombinant protein was tested through the expression of the positive recombinant with different time span and different IPTG concentration. Lots of the protein of DuIFN-αwere expressed in BL21(DE3)plysS with 1 mmol·L-1 IPTG for 4 hours and its molecular weight for 34 000.展开更多
基金Studying Abroad and Coming Back Home Fund (LC02C08).
文摘To obtain the protein of duck interferon alpha and study its biological activities, the prokaryotic expression vector of DuIFN-αwas constructed and expressed in BL21 (DE3) plysS. Using PCR technique, the protein gene of DuIFN-αwas cloned from pMD-18-duIFN-αrecombinant. The gene was then inserted to pGEM-T vector and identified by restriction endonuclease analysis and sequencing. DuIFN-αwas ligated with the prokaryotic expression vector of pET30 a, then transformed into BL21 (DE3) plysS. The best inducing time and IPTG concentration for the expression of this recombinant protein was tested through the expression of the positive recombinant with different time span and different IPTG concentration. Lots of the protein of DuIFN-αwere expressed in BL21(DE3)plysS with 1 mmol·L-1 IPTG for 4 hours and its molecular weight for 34 000.