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H9N2亚型禽流感病毒糖基化位点突变株的构建与生物学特性研究 被引量:6
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作者 廖昌韬 曾凡桂 +4 位作者 严专强 覃健萍 曹永长 谢青梅 陈峰 《动物医学进展》 北大核心 2018年第7期18-22,共5页
为了解H9N2亚型禽流感病毒(AIV)HA蛋白第200位N-糖基化位点的功能,利用流感病毒8质粒反向遗传系统拯救出重组病毒r-WZP株(缺失HA基因第200位N-糖基化位点)和r-WZP200株(含有HA基因第200位N-糖基化位点)。两株病毒的内部基因来自病毒株A/... 为了解H9N2亚型禽流感病毒(AIV)HA蛋白第200位N-糖基化位点的功能,利用流感病毒8质粒反向遗传系统拯救出重组病毒r-WZP株(缺失HA基因第200位N-糖基化位点)和r-WZP200株(含有HA基因第200位N-糖基化位点)。两株病毒的内部基因来自病毒株A/Puerto Rico/8/34(H1N1),HA基因与NA基因均来自H9N2AIV流行毒株。结果表明,两株病毒均能够在SPF鸡胚中稳定增殖,这一位点的缺失致使病毒与鸡红细胞的结合能力下降,HA效价热稳定性提高,对SPF鸡胚的致死能力增强,对进一步研究H9N2亚型AIV HA蛋白N-糖基化位点的功能具有重要的参考价值。 展开更多
关键词 H9N2亚型禽流感病毒 HA基因 N-糖基化位点
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Sequence and phylogenetic analysis of hemagglutinin genes of H9N2 influenza viruses isolated from chicken in China from 2013 to 2015 被引量:5
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作者 SU Xiao-na XIE Qing-mei +4 位作者 LIAO Chang-tao yan zhuan-qiang CHEN Wei-guo BI Ying-zuo CHEN Feng 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2016年第11期2604-2612,共9页
H9N2 avian influenza virus(AIV) infection is a major problem in poultry industry worldwide. In this study, molecular characterizations and phylogenetic relationships of hemagglutinin(HA) gene sequences of H9N2 AIV... H9N2 avian influenza virus(AIV) infection is a major problem in poultry industry worldwide. In this study, molecular characterizations and phylogenetic relationships of hemagglutinin(HA) gene sequences of H9N2 AIV of 5 Chinese isolates in 2014 recently available in Gen Bank, 3 widely used vaccine strains, and 52 novel isolates in China from 2013 to 2015 were analyzed. The homology analysis showed that the nucleotide sequences of HA gene of these recent Chinese H9N2 AIV isolates shared homologies from 94.1 to 99.9%. Phylogenetic analysis showed that all isolates belonged to AIV lineage h9.4.2.5. Fifty-six out of the 57 recent Chinese H9N2 AIV isolates had the motifs PSRSSR↓GLF at the cleavage sites within the HA protein, while one isolate PWH01 harbored LSRSSR↓GLF. Remarkably, all of the recent Chinese H9N2 AIV strains had the Q216 L substitution in the receptor binding site, which indicated that they had potential to infect humans. Most of recent Chinese H9N2 AIV isolates lost the potential N-linked glycosylation site at residues 200–202 compared with vaccine strains. This present study demonstrated that AIV lineage h9.4.2.5 was more predominant in China than other lineages as it harbored all the H9N2 AIV isolated between 2013 and 2015. Also we showed the importance of continuous surveillance of emerging H9N2 AIV in China and update of vaccine formulation accordingly in order to prevent and control H9N2 AIV. 展开更多
关键词 avian influenza virus H9N2 subtype phylogenetic analysis hemagglutinin gene
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生物反应器悬浮培养禽偏肺病毒制备弱毒疫苗工艺的优化
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作者 蒋天华 周庆丰 +1 位作者 严专强 周展 《中国生物制品学杂志》 CAS CSCD 北大核心 2022年第3期316-320,共5页
目的对生物反应器悬浮培养禽偏肺病毒(avian metapneumovirus,aMPV)制备弱毒疫苗的工艺进行优化。方法采用分批培养的方法优化悬浮BHK-21细胞在生物反应器内培养及aMPV增殖工艺,并对制备的疫苗进行动物安全性及攻毒保护试验。结果采用1.... 目的对生物反应器悬浮培养禽偏肺病毒(avian metapneumovirus,aMPV)制备弱毒疫苗的工艺进行优化。方法采用分批培养的方法优化悬浮BHK-21细胞在生物反应器内培养及aMPV增殖工艺,并对制备的疫苗进行动物安全性及攻毒保护试验。结果采用1.0×10^(6)cells/mL的初始接种密度培养BHK-21悬浮细胞,细胞能快速度过延滞期进入对数生长期,培养72 h细胞密度可达1.6×10^(7)cells/mL。调整细胞密度为3.0×10^(6)cells/mL,按照感染复数MOI=0.1的剂量接种病毒,病毒含量可达10^(8.67)TCID_(50)/mL。制备的疫苗安全性良好,可有效保护番鸭,保护率为100%。结论优化了生物反应器培养aMPV的工艺,可获得较高的病毒效价,为aMPV疫苗的规模化生产提供了参考。 展开更多
关键词 BHK-21细胞 禽偏肺病毒 悬浮培养 生物反应器
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