Ducks inoculated intravenously or via the ocular-nasal-oral-cloacal routes with a highly pathogenic avian influenza virus,A/duck/Guangdong/220/2004(H5N1),developed systemic hyperemia,congestion,hemorrhage,thrombosis a...Ducks inoculated intravenously or via the ocular-nasal-oral-cloacal routes with a highly pathogenic avian influenza virus,A/duck/Guangdong/220/2004(H5N1),developed systemic hyperemia,congestion,hemorrhage,thrombosis and edema in various organs,as well as necrosis or apoptosis in the parenchyma of the heart,liver,spleen,lungs,kidneys,pancreas,brain,thymus and bursa of Fabricius.The main manifestations were angiitis,necrotic pancreatitis,atrophic necrotic thymitis and bursitis Fabricii,splenitis,tracheitis,hemorrhagic bronchointerstitial pneumonia,viral myocarditis,nonsuppurative encephalitis,focal viral hepatitis,ulcerative enteritis,renal tubule interstitial nephritis,and intraglomerular mesangial cell hyperplastic glomerular nephritis.The results demonstrated that the mechanism of pathogenesis involved cellular necrosis and apoptosis,and that death of the ducks was caused by severe pathologic trauma occurring in multiple visceral organs.展开更多
To study biological activities of Duck Interferon Alpha （DuIFN-α） and prepare antivirus medicine, the eukaryotic expression vector of mature polypeptide of Duck Interferon Alpha （mDuIFN-α） gene was constructed a...To study biological activities of Duck Interferon Alpha （DuIFN-α） and prepare antivirus medicine, the eukaryotic expression vector of mature polypeptide of Duck Interferon Alpha （mDuIFN-α） gene was constructed and expressed in insect cell. By means of PCR technique, the mDuIFN-α gene was cloned from pMD-18-duIFN-α recombinant_ The gene was then inserted to pGEM-T vector and identified by restriction endonuclease analysis and sequencing The mDuIFN-α gene was ligated with the eukaryotic expression vector pMelBacA. then transfected into Sf9 cell line. Recombinant polypeptide was effectively expressed in insect cell and its molecular weight was 34 ku.展开更多
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supported by the Guangdong Province Science&Technology Hard Nut Project(2004A2090102)Guangdong Province Education Bureau Science Foundation Project(Z02003)
文摘Ducks inoculated intravenously or via the ocular-nasal-oral-cloacal routes with a highly pathogenic avian influenza virus,A/duck/Guangdong/220/2004(H5N1),developed systemic hyperemia,congestion,hemorrhage,thrombosis and edema in various organs,as well as necrosis or apoptosis in the parenchyma of the heart,liver,spleen,lungs,kidneys,pancreas,brain,thymus and bursa of Fabricius.The main manifestations were angiitis,necrotic pancreatitis,atrophic necrotic thymitis and bursitis Fabricii,splenitis,tracheitis,hemorrhagic bronchointerstitial pneumonia,viral myocarditis,nonsuppurative encephalitis,focal viral hepatitis,ulcerative enteritis,renal tubule interstitial nephritis,and intraglomerular mesangial cell hyperplastic glomerular nephritis.The results demonstrated that the mechanism of pathogenesis involved cellular necrosis and apoptosis,and that death of the ducks was caused by severe pathologic trauma occurring in multiple visceral organs.
文摘To study biological activities of Duck Interferon Alpha （DuIFN-α） and prepare antivirus medicine, the eukaryotic expression vector of mature polypeptide of Duck Interferon Alpha （mDuIFN-α） gene was constructed and expressed in insect cell. By means of PCR technique, the mDuIFN-α gene was cloned from pMD-18-duIFN-α recombinant_ The gene was then inserted to pGEM-T vector and identified by restriction endonuclease analysis and sequencing The mDuIFN-α gene was ligated with the eukaryotic expression vector pMelBacA. then transfected into Sf9 cell line. Recombinant polypeptide was effectively expressed in insect cell and its molecular weight was 34 ku.
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.
