一株野鸭源禽流感病毒NS基因的分子克隆和测序

Molecular Cloning and Sequencing of the NS Gene of an Avian Influenza Virus Isolated from Wild Ducks

根据Genbank上已发表的H4N6亚型的NS基因序列,设计一对特异性引物,成功扩增出一株野鸭源禽流感病毒扎龙分离株A/mallard/ZhaLong/88(H4N6)的NS基因,并将该片段连接到PMD18-T载体上,连接产物转化至DH5α感受态细胞,对目的片段进行测序,获得了NS基因全序列,大小为887bp。该片段的核酸序列与已知的几个毒株序列进行同源性比较,同源性为78.4%～97.6%,推导的氨基酸序列同源性为80.8%～98.6%。

A pair of primers was designed according to the NS gene sequence of H4N6 subtype avian influenza viruses published in GcnBank, and then NS gene of an avian influenza virus isolated in wild ducks from Zhalang Natural Reserve was amplified successfully. The desired fragment was inserted into pMD18-T vector, and the recombinant plasmid was transformed into DHSα competent cell, then the sequence of 887 bp of NS gene was gained. The nucleotide and deduced amino acid sequence of NS1 gene of this isolate was compared with some published NS sequence. Result showed that the homologies of nucleotide and amino acid were 78.4% ～ 97.6% and 80.8% ～ 98.6%, respectively.