The small envelope protein （E） gene of avian infectious bronchitis virus （IBV） M41 strain was cloned, and then it was subcloned into prokaryotic expressing vector pGEX-6P-1. The recombinant plasmid was transformed...The small envelope protein （E） gene of avian infectious bronchitis virus （IBV） M41 strain was cloned, and then it was subcloned into prokaryotic expressing vector pGEX-6P-1. The recombinant plasmid was transformed into E.coli. BL21 and induced by IPTG. SDS-PAGE result showed that when objective protein fused with GST （about 20 ku）, the relative molecular mass of fusion protein was 38 ku. It indicated that objective protein was about 12.4 ku. The result showed that E protein was expressed successfully, it was useful to the subsequent E protein research.展开更多
Infectious bronchitis(IB) is a highly contagious avian disease caused by infection with infectious bronchitis virus(IBV),which seriously affects the development of the global poultry industry. The distribution of TW I...Infectious bronchitis(IB) is a highly contagious avian disease caused by infection with infectious bronchitis virus(IBV),which seriously affects the development of the global poultry industry. The distribution of TW I-type IBV in China has increased in recent years, becoming a widespread genotype. We previously isolated a TW I-type IBV strain termed CK/CH/GD/GZ14 in 2014, but its pathogenicity and possibility for vaccine development were not explored. Therefore, this research aimed to develop a live-attenuated virus vaccine based on the CK/CH/GD/GZ14 strain. The wild type IBV CK/CH/GD/GZ14 strain was serially passaged in SPF embryos for 145 generations. The morbidity and mortality rate of wildtype strain in 14 day-old chickens is 100% and 80% respectively, while the morbidity rate in the attenuated strain was 20%in the 95 th and 105 th generations and there was no death. Histopathological observations showed that the pathogenicity of the 95th and 105th generations in chickens was significantly weakened. Further challenge experiments confirmed that the attenuated CK/CH/GD/GZ14 strain in the 95th and 105 th generations could resist CK/CH/GD/GZ14(5th generation)infection and the protection rate was 80%. Tracheal cilia stagnation, virus shedding, and viral load experiments confirmed that the 95 th and 105th generations provide good immune protection in chickens, and the immunogenicity of the 105th generation is better than that of the 95th generation. These data suggest that the attenuated CK/CH/GD/GZ14 strain in the105th generation may be applied as a vaccine candidate against TW I-type IBV.展开更多
Dear Editor,Infectious bronchitis(IB),one of the most common and difficult poultry diseases,is caused by a gammacoronavirus named infectious bronchitis virus(IBV).IBV frequently causes respiratory and/or renal disease...Dear Editor,Infectious bronchitis(IB),one of the most common and difficult poultry diseases,is caused by a gammacoronavirus named infectious bronchitis virus(IBV).IBV frequently causes respiratory and/or renal diseases in chickens and egg production losses in hens.IB has展开更多
Acording to Avian Infectious Brochitis virus( IBV) Beaudette strain S1 gene sequence, a pair of primers were designed and synthesized. With the primers , IBV Guangdong isolation strain GD05 S1 gene was successively am...Acording to Avian Infectious Brochitis virus( IBV) Beaudette strain S1 gene sequence, a pair of primers were designed and synthesized. With the primers , IBV Guangdong isolation strain GD05 S1 gene was successively amplified by RT-PCR.PCR product was digested with BstY I,\%Hae\% III and \%Pst\% I respectively, the result showed RFLP pattern of was the same as that of M41 S1 gene. IBV GD05 strain was thought as Mass serotype primaryly. IBV GD05 S1 gene was cloned into pGEM-T vector and sequenced, its sequence was consisted of 1611 base pairs. By comparison , the nucleotide sequence was 97.14% identical to that of IBV M41. IBV GD05 S1 gene was subcloned into expression vector pET21d. SDS-PAGE experiment showed that it expressed in \%E.coli.\%展开更多
In this study, the genes encoding main structural proteins and 3′ untranslated region (UTR) of a commercial domestic vaccine strain H52 and a commercial foreign vaccine strain H52 of infectious bronchitis virus (IBV)...In this study, the genes encoding main structural proteins and 3′ untranslated region (UTR) of a commercial domestic vaccine strain H52 and a commercial foreign vaccine strain H52 of infectious bronchitis virus (IBV) were obtained by RT-PCR, then were cloned and sequenced respectively. The sequence analyses showed the genetic information of S1 gene, M gene, N gene and 3′UTR of domestic vaccine strain H52 was remarkably different from that of foreign H52 strain and that of published sequences of H52 strain. Compared with foreign H52 strain, this domestic H52 strain was genetically closer with M41 strain and was located in the same branch of phylogenetic tree based on the sequences of main structural protein genes. The results strongly suggested that it was necessary to build genetic information archives of seed stock of IBV vaccine strains.展开更多
文摘The small envelope protein （E） gene of avian infectious bronchitis virus （IBV） M41 strain was cloned, and then it was subcloned into prokaryotic expressing vector pGEX-6P-1. The recombinant plasmid was transformed into E.coli. BL21 and induced by IPTG. SDS-PAGE result showed that when objective protein fused with GST （about 20 ku）, the relative molecular mass of fusion protein was 38 ku. It indicated that objective protein was about 12.4 ku. The result showed that E protein was expressed successfully, it was useful to the subsequent E protein research.
基金This study was supported by the Key Research and Development Program of Guangdong Province(2020B020222001)the Construction of Modern Agricultural Science and Technology Innovation Alliance in Guangdong Province(2020KJ128)+5 种基金the Natural Science Foundation of Guangdong Province(2019A1515012006)the National Natural Science Foundation of China(31902252)the Special Project of National Modern Agricultural Industrial Technology System(CARS-41)the National Modern Agricultural Industry Science and Technology Innovation Center in Guangzhou(2018kczx01)the National Key R&D Program of China(2017YFD0502001)the Creation of a Triple Chimeric Vaccine(rIBV-ND-H9)Using Avian Infectious Bronchitis Attenuated D90 as a Vector(2017KZDM008)。
文摘Infectious bronchitis(IB) is a highly contagious avian disease caused by infection with infectious bronchitis virus(IBV),which seriously affects the development of the global poultry industry. The distribution of TW I-type IBV in China has increased in recent years, becoming a widespread genotype. We previously isolated a TW I-type IBV strain termed CK/CH/GD/GZ14 in 2014, but its pathogenicity and possibility for vaccine development were not explored. Therefore, this research aimed to develop a live-attenuated virus vaccine based on the CK/CH/GD/GZ14 strain. The wild type IBV CK/CH/GD/GZ14 strain was serially passaged in SPF embryos for 145 generations. The morbidity and mortality rate of wildtype strain in 14 day-old chickens is 100% and 80% respectively, while the morbidity rate in the attenuated strain was 20%in the 95 th and 105 th generations and there was no death. Histopathological observations showed that the pathogenicity of the 95th and 105th generations in chickens was significantly weakened. Further challenge experiments confirmed that the attenuated CK/CH/GD/GZ14 strain in the 95th and 105 th generations could resist CK/CH/GD/GZ14(5th generation)infection and the protection rate was 80%. Tracheal cilia stagnation, virus shedding, and viral load experiments confirmed that the 95 th and 105th generations provide good immune protection in chickens, and the immunogenicity of the 105th generation is better than that of the 95th generation. These data suggest that the attenuated CK/CH/GD/GZ14 strain in the105th generation may be applied as a vaccine candidate against TW I-type IBV.
基金supported in part through BattelleMemorial Institute’s prime contract with the US National Institute of Allergy and Infectious Diseases(NIAID)under Contract No.HHSN272200700016I
文摘Dear Editor,Infectious bronchitis(IB),one of the most common and difficult poultry diseases,is caused by a gammacoronavirus named infectious bronchitis virus(IBV).IBV frequently causes respiratory and/or renal diseases in chickens and egg production losses in hens.IB has
文摘Acording to Avian Infectious Brochitis virus( IBV) Beaudette strain S1 gene sequence, a pair of primers were designed and synthesized. With the primers , IBV Guangdong isolation strain GD05 S1 gene was successively amplified by RT-PCR.PCR product was digested with BstY I,\%Hae\% III and \%Pst\% I respectively, the result showed RFLP pattern of was the same as that of M41 S1 gene. IBV GD05 strain was thought as Mass serotype primaryly. IBV GD05 S1 gene was cloned into pGEM-T vector and sequenced, its sequence was consisted of 1611 base pairs. By comparison , the nucleotide sequence was 97.14% identical to that of IBV M41. IBV GD05 S1 gene was subcloned into expression vector pET21d. SDS-PAGE experiment showed that it expressed in \%E.coli.\%
文摘In this study, the genes encoding main structural proteins and 3′ untranslated region (UTR) of a commercial domestic vaccine strain H52 and a commercial foreign vaccine strain H52 of infectious bronchitis virus (IBV) were obtained by RT-PCR, then were cloned and sequenced respectively. The sequence analyses showed the genetic information of S1 gene, M gene, N gene and 3′UTR of domestic vaccine strain H52 was remarkably different from that of foreign H52 strain and that of published sequences of H52 strain. Compared with foreign H52 strain, this domestic H52 strain was genetically closer with M41 strain and was located in the same branch of phylogenetic tree based on the sequences of main structural protein genes. The results strongly suggested that it was necessary to build genetic information archives of seed stock of IBV vaccine strains.
