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
For developing efficient vaccines, it is essential to identify which amino acid changes are most important to the survival of the virus. We investigate the amino acid substitution features in the Avian Infectious Bron...For developing efficient vaccines, it is essential to identify which amino acid changes are most important to the survival of the virus. We investigate the amino acid substitution features in the Avian Infectious Bronchitis Virus （AIBV） antigenic domain of a vaccine serotype （DE072） and a virulent viral strain （GA98） to better understand adaptive evolution of AIBV. In addition, the SARS Coronavirus （SARS-CoV） was also analyzed in the same way. It is interesting to find that extreme comparability exists between AIBV and SARS in amino acid substitution pattern. It suggests that amino acid changes that result in overall shift of residue charge and polarity should be paid special attention to during the development of vaccines.展开更多
[ Objective] The aim was to isolate and identify avian infectious bronchitis virus （IBV） from diseased chickens. [ Method] IBVs were iso- lated from the diseased chickens in a chicken farm in Anhui Province with bli...[ Objective] The aim was to isolate and identify avian infectious bronchitis virus （IBV） from diseased chickens. [ Method] IBVs were iso- lated from the diseased chickens in a chicken farm in Anhui Province with blind passage method to observe virus pathogenicity. Then animal regres- sion test was made to replicate symptoms of bronchial congestion in SPF chickens and S1 gene segments were amplified and isolated, followed by comparison with IBV vaccine strains. [ Result] Detection of Hemagglutinating activity （HA） showed that allantoic fluid had no concerning effect on erythrocyte, suggesting that NDV and AIV were not included in the isolated viruses. However, the erythrocyte could be agglutinated with allantoic fluid treated with 1% of pancreatin, which is in consistent with biological characters of IBV. After SPF chickens were inoculated with the 6^th SPF al- lantoic fluid, bronchial congestion was replicated, proving that the isolated virus was avian IBV, named IBV XZ strain. [ Conclusion] This study pro- vides a theoretical basis for prevention of avian infectious bronchitis.展开更多
[Objective] To study the prokaryotic expression and antigenicity identification of S1 gene from avian infectious bronchitis virus （IBV）. [Method] The S1 gene was cloned into a pMD18-T vector to yield a recombinant p...[Objective] To study the prokaryotic expression and antigenicity identification of S1 gene from avian infectious bronchitis virus （IBV）. [Method] The S1 gene was cloned into a pMD18-T vector to yield a recombinant plasmids pMD18-T-IBV-S1. Then S1 gene was inserted into the multiple cloning site of a prokaryotic expression vector pET-32a （ ＋ ）. The recombinant plasmid was transformed into E. coil BL21. The recombinant protein was induced by IPTG and measured by SDS-PAGE and western-blotting. [Result] The S1 gene was successfully expressed in E. coil BL21, the fusion proteins were about 66.0 kDa in a form of inclusion body. Western-blotting test showed that the recombinant proteins could be identified by IBV polyclonal antibody. [ Conclusion] The recombinant proteins of S1 gene have the antigenicity, which lays a good foundation for further research on new generation vaccine of IBV.展开更多
[ Objective] The aim was to to establish a kind of peculiar, sensitive and quick fluorescent PCR detection method. [Method] A peculiar, sensitive and quick method of fluorescent PCR detection for avian infectious bron...[ Objective] The aim was to to establish a kind of peculiar, sensitive and quick fluorescent PCR detection method. [Method] A peculiar, sensitive and quick method of fluorescent PCR detection for avian infectious bronchitis virus was established, the standard curve was built, specific primers, susceptibility and repeatability was detected. [ Result] This method diagnosed avian infectious bronchitis virus peculiarly, sensitively and quickly, simple and easy to use, time short, suitable for clinical testing. [ Conclusion] This research laid the foundation to diagnose avian infectious bronchitis virus.展开更多
A novel proventriculopathogic variant (isolate ZJ971) of infectious bronchitis virus (IBV) was identified from enlarged proeventriculus of the sick chickens in the study. The S gene cDNA segment with 3.6 kb in length ...A novel proventriculopathogic variant (isolate ZJ971) of infectious bronchitis virus (IBV) was identified from enlarged proeventriculus of the sick chickens in the study. The S gene cDNA segment with 3.6 kb in length was amplified by RT-PCR with special primers from the ZJ971 viral isolate of (IBV) and cloned into plasmid pBluescript SK( + ). The recombinants containing S gene of IBV-ZJ971 isolate were identified by digestion of restriction enzyme EcoRI, BamHI and PCR amplification. The cloned S gene from isolate IBV-7J971 was composed of 3492 bp in length encoding for a polypeptide of 1080 amino acids. Comparing the nucleotide of S gene of IBV isolate ZJ971 with that of reported IBV strains Beaudette, M41, Ark99 and CuT2, the homology was 97.3%, 97.5%, 88.6% and 85.6%, respectively; and the homology of the deduced amino acids of S protein of IBV isolate ZJ971 was 96%, 96.3%, 86.1% and 83.1% respectively; especially, the mutation of 3241st nucleotide of S gene of IBV isolate ZJ971 from G to T resulted in the translating termination of S protein at 3240th nucleotide site.展开更多
根据发表的鸡传染性支气管炎病毒N基因序列设计一对引物,经RT-PCR扩增得到1 230 bp DNA片段,将其克隆到表达载体pET-28a(+)中,构建重组质粒pET-N,经测序鉴定正确后,将其转化感受态细胞E.coliBL21(DE3),并进行IPTG诱导表达,结果表明,重...根据发表的鸡传染性支气管炎病毒N基因序列设计一对引物,经RT-PCR扩增得到1 230 bp DNA片段,将其克隆到表达载体pET-28a(+)中,构建重组质粒pET-N,经测序鉴定正确后,将其转化感受态细胞E.coliBL21(DE3),并进行IPTG诱导表达,结果表明,重组菌可表达分子质量为50 kD的融合蛋白。Western-blotting结果显示,该重组蛋白能与IBV阳性血清发生特异性的免疫印迹反应,表明该重组蛋白具有良好的免疫原性。展开更多
Purified avian infectious bronchitis virus (IBV) was used to screen a random phage dis-play peptide library. After the fourth panning, 10 positive phages were sequenced and characterized. The phages specifically inhib...Purified avian infectious bronchitis virus (IBV) was used to screen a random phage dis-play peptide library. After the fourth panning, 10 positive phages were sequenced and characterized. The phages specifically inhibited IBV infectivity in HeLa cells and blocked IBV haemagglutination. One linear peptide “GSH HRH VHS PFV” from the positive phages with the highest neutralization titer was synthesized and this peptide inhibited IBV infection in HeLa as well. The results may contribute to development of antiviral therapeutics for IBV and studying the determinants for viral and cell interac-tion.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
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.
文摘For developing efficient vaccines, it is essential to identify which amino acid changes are most important to the survival of the virus. We investigate the amino acid substitution features in the Avian Infectious Bronchitis Virus （AIBV） antigenic domain of a vaccine serotype （DE072） and a virulent viral strain （GA98） to better understand adaptive evolution of AIBV. In addition, the SARS Coronavirus （SARS-CoV） was also analyzed in the same way. It is interesting to find that extreme comparability exists between AIBV and SARS in amino acid substitution pattern. It suggests that amino acid changes that result in overall shift of residue charge and polarity should be paid special attention to during the development of vaccines.
文摘[ Objective] The aim was to isolate and identify avian infectious bronchitis virus （IBV） from diseased chickens. [ Method] IBVs were iso- lated from the diseased chickens in a chicken farm in Anhui Province with blind passage method to observe virus pathogenicity. Then animal regres- sion test was made to replicate symptoms of bronchial congestion in SPF chickens and S1 gene segments were amplified and isolated, followed by comparison with IBV vaccine strains. [ Result] Detection of Hemagglutinating activity （HA） showed that allantoic fluid had no concerning effect on erythrocyte, suggesting that NDV and AIV were not included in the isolated viruses. However, the erythrocyte could be agglutinated with allantoic fluid treated with 1% of pancreatin, which is in consistent with biological characters of IBV. After SPF chickens were inoculated with the 6^th SPF al- lantoic fluid, bronchial congestion was replicated, proving that the isolated virus was avian IBV, named IBV XZ strain. [ Conclusion] This study pro- vides a theoretical basis for prevention of avian infectious bronchitis.
基金supported by Science and Technology Star Project of Beijing (2005B35)
文摘[Objective] To study the prokaryotic expression and antigenicity identification of S1 gene from avian infectious bronchitis virus （IBV）. [Method] The S1 gene was cloned into a pMD18-T vector to yield a recombinant plasmids pMD18-T-IBV-S1. Then S1 gene was inserted into the multiple cloning site of a prokaryotic expression vector pET-32a （ ＋ ）. The recombinant plasmid was transformed into E. coil BL21. The recombinant protein was induced by IPTG and measured by SDS-PAGE and western-blotting. [Result] The S1 gene was successfully expressed in E. coil BL21, the fusion proteins were about 66.0 kDa in a form of inclusion body. Western-blotting test showed that the recombinant proteins could be identified by IBV polyclonal antibody. [ Conclusion] The recombinant proteins of S1 gene have the antigenicity, which lays a good foundation for further research on new generation vaccine of IBV.
基金Funds for the Central Universities of Dalian Nationalities University (DC12010304)
文摘[ Objective] The aim was to to establish a kind of peculiar, sensitive and quick fluorescent PCR detection method. [Method] A peculiar, sensitive and quick method of fluorescent PCR detection for avian infectious bronchitis virus was established, the standard curve was built, specific primers, susceptibility and repeatability was detected. [ Result] This method diagnosed avian infectious bronchitis virus peculiarly, sensitively and quickly, simple and easy to use, time short, suitable for clinical testing. [ Conclusion] This research laid the foundation to diagnose avian infectious bronchitis virus.
基金the National NatureScience Foundation of China(30070570)the NatureScience Foundation of Zhejiang Province(399411) the Science and Technology Commission of Zhejiang Province(991102030).
文摘A novel proventriculopathogic variant (isolate ZJ971) of infectious bronchitis virus (IBV) was identified from enlarged proeventriculus of the sick chickens in the study. The S gene cDNA segment with 3.6 kb in length was amplified by RT-PCR with special primers from the ZJ971 viral isolate of (IBV) and cloned into plasmid pBluescript SK( + ). The recombinants containing S gene of IBV-ZJ971 isolate were identified by digestion of restriction enzyme EcoRI, BamHI and PCR amplification. The cloned S gene from isolate IBV-7J971 was composed of 3492 bp in length encoding for a polypeptide of 1080 amino acids. Comparing the nucleotide of S gene of IBV isolate ZJ971 with that of reported IBV strains Beaudette, M41, Ark99 and CuT2, the homology was 97.3%, 97.5%, 88.6% and 85.6%, respectively; and the homology of the deduced amino acids of S protein of IBV isolate ZJ971 was 96%, 96.3%, 86.1% and 83.1% respectively; especially, the mutation of 3241st nucleotide of S gene of IBV isolate ZJ971 from G to T resulted in the translating termination of S protein at 3240th nucleotide site.
文摘根据发表的鸡传染性支气管炎病毒N基因序列设计一对引物,经RT-PCR扩增得到1 230 bp DNA片段,将其克隆到表达载体pET-28a(+)中,构建重组质粒pET-N,经测序鉴定正确后,将其转化感受态细胞E.coliBL21(DE3),并进行IPTG诱导表达,结果表明,重组菌可表达分子质量为50 kD的融合蛋白。Western-blotting结果显示,该重组蛋白能与IBV阳性血清发生特异性的免疫印迹反应,表明该重组蛋白具有良好的免疫原性。
文摘Purified avian infectious bronchitis virus (IBV) was used to screen a random phage dis-play peptide library. After the fourth panning, 10 positive phages were sequenced and characterized. The phages specifically inhibited IBV infectivity in HeLa cells and blocked IBV haemagglutination. One linear peptide “GSH HRH VHS PFV” from the positive phages with the highest neutralization titer was synthesized and this peptide inhibited IBV infection in HeLa as well. The results may contribute to development of antiviral therapeutics for IBV and studying the determinants for viral and cell interac-tion.
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
