摘要
利用反向遗传操作技术验证了H9N2亚型禽流感病毒流行株(A/Chicken/Shandong/903,简称903)HA蛋白200和295位氨基酸糖基化可以影响病毒对α-2,6唾液酸受体的亲和性,同时也能影响病毒在小鼠体内复制能力。固相ELISA结果显示,突变病毒N200Q(N→Q)和N295Q(N→Q)都能降低病毒对α-2,6唾液酸受体的亲和性。荧光定量结果显示,突变病毒N295Q主要在小鼠肺脏、脾脏和肾脏中复制(M基因拷贝分别为903野毒的56,64,39倍);而突变病毒N200Q主要在小鼠肾脏中复制(M基因拷贝分别为903野毒的936倍)。本试验为阐明H9N2亚型流感病毒跨种间传播的分子机制提供依据。
Reverse genetic manipulation technology was used to determine whether H9N2 subtype avian influenza virus strain(A/Chicken/Shandong/903,903)HA protein 200 and 295amino acid glycosylation could affect the affinity of virus toα-2,6sialic acid receptor,as well as its replication ability in mice.Solid-phase ELISA result showed that mutant viruses N200Q(N→Q)and N295Q(N→Q)could reduce the affinity of virus toα-2,6sialic acid receptor.Fluorescent quantitative results showed that mutant virus N295 Q mainly replicated in the lung,spleen,and kidney of mice(M gene copy was 56-,64-,and 39-fold of 903 wild virus,respectively),and mutant virus N200 Q mainly replicated in the kidney(M gene copy was 936-fold of 903 wild virus).The research results provided an important theoretical and experimental basis for explaining the molecular mechanism of H9N2 subtype avian influenza virus in interspecies transmission.
引文
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