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
Avian infectious bronchitis(IB)is an acute and highly contagious disease caused by infectious bronchitis virus(IBV).In the study,according to IBV gene sequences published in Gen Bank,specific primers were designed to ...Avian infectious bronchitis(IB)is an acute and highly contagious disease caused by infectious bronchitis virus(IBV).In the study,according to IBV gene sequences published in Gen Bank,specific primers were designed to clone N gene by RT-PCR,and this gene was inserted into p ET-30a(+)vector resulting in a prokaryotic expression plasma p ET-30a-N.The results of SDS-PAGE and Western Blot analysis showed that the recombinant protein was expressed successfully and had good reactivity with IBV positive serum.Using purified recombinant N protein as a coating antigen,the indirect ELISA protocol was established and optimized,in which N protein was 2.5μg·m L^-1 of concentration,sample serum of 1:40 dilution.For clinical specimen,the IBV antibodies could be detected by this method efficiently and got nearly the same results as those of IBV-mediated ELISA.It would provide a good tool for rapid diagnosis and epidemiological study of avian infectious bronchitis.展开更多
CAP,an adenylyl cyclase-associated protein,is predicted to be involved in cytoskeletal organization and signal transduction.Recently,we found that CAP may play an important role in fuzz-like fiber cell initiation in c...CAP,an adenylyl cyclase-associated protein,is predicted to be involved in cytoskeletal organization and signal transduction.Recently,we found that CAP may play an important role in fuzz-like fiber cell initiation in cotton.For the further research,we isolated two CAP homologues from wild展开更多
The zinc finger proteins belong to the largest family of transcription factors.But there is little research of Cys2/His2 type zinc finger proteins in cotton,and there is no submission of correlating
The capsid protein precursor （P1）, which plays a major role for the generation of polypeptides of swine vesicular disease virus （SVDV）, was cloned from SVDV HK/70 strain into the retroviral vector pBABE puro and e...The capsid protein precursor （P1）, which plays a major role for the generation of polypeptides of swine vesicular disease virus （SVDV）, was cloned from SVDV HK/70 strain into the retroviral vector pBABE puro and expressed in the mammalian cell line PK15 through the retroviral expression system. The activity of recombinant protein to induce immune response was evaluated in guinea pigs. IFA and Western Blot were used to detect the recombinant protein expression. The results showed that the recombinant protein could be recognized by SVDV positive serum, and animal test showed SVDV-specific antibodies. All of those results indicate that a retroviral-based vaccine carrying the capsid protein precursor （P1） of SVD is able to be expressed in the eukaryotic cell and elicites strong SVDV-specific immune responses in guinea pigs.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Supported by the National Natural Science Foundation of China(31172295,31272569)。
文摘Avian infectious bronchitis(IB)is an acute and highly contagious disease caused by infectious bronchitis virus(IBV).In the study,according to IBV gene sequences published in Gen Bank,specific primers were designed to clone N gene by RT-PCR,and this gene was inserted into p ET-30a(+)vector resulting in a prokaryotic expression plasma p ET-30a-N.The results of SDS-PAGE and Western Blot analysis showed that the recombinant protein was expressed successfully and had good reactivity with IBV positive serum.Using purified recombinant N protein as a coating antigen,the indirect ELISA protocol was established and optimized,in which N protein was 2.5μg·m L^-1 of concentration,sample serum of 1:40 dilution.For clinical specimen,the IBV antibodies could be detected by this method efficiently and got nearly the same results as those of IBV-mediated ELISA.It would provide a good tool for rapid diagnosis and epidemiological study of avian infectious bronchitis.
文摘CAP,an adenylyl cyclase-associated protein,is predicted to be involved in cytoskeletal organization and signal transduction.Recently,we found that CAP may play an important role in fuzz-like fiber cell initiation in cotton.For the further research,we isolated two CAP homologues from wild
文摘The zinc finger proteins belong to the largest family of transcription factors.But there is little research of Cys2/His2 type zinc finger proteins in cotton,and there is no submission of correlating
基金Supported by Key Technology R&D Programme (2006BAD06A03)
文摘The capsid protein precursor （P1）, which plays a major role for the generation of polypeptides of swine vesicular disease virus （SVDV）, was cloned from SVDV HK/70 strain into the retroviral vector pBABE puro and expressed in the mammalian cell line PK15 through the retroviral expression system. The activity of recombinant protein to induce immune response was evaluated in guinea pigs. IFA and Western Blot were used to detect the recombinant protein expression. The results showed that the recombinant protein could be recognized by SVDV positive serum, and animal test showed SVDV-specific antibodies. All of those results indicate that a retroviral-based vaccine carrying the capsid protein precursor （P1） of SVD is able to be expressed in the eukaryotic cell and elicites strong SVDV-specific immune responses in guinea pigs.
