从人分离出两株甲型流感H9N2亚型毒株内部基因特性的研究

Characterization of internal genes of two strains of influenza A (H9N2) virus isolated from men

目的 了解 2株从人分离出的H9N2亚型毒株内部基因特性 ,并弄清其来源。方法 用RT PCR扩增目的基因 ,用PGEM T Vector (美国Promega公司 ) ,4℃过夜连接 ,重组质粒转入dH5α细菌 ,筛选阳性菌落 ,酶切鉴定 ,送六合通公司自动测序。然后进行进化树分析。结果 2株测定毒株内部基因均为G9基因系 ,它们相互间除PA基因有差异外 ,其余 5个基因均相同。结论 2株测定毒株的基因组均为G9基因系 ,它们是由携带不同基因特性H9N2毒株的禽群分别直接感染人 。

Objective To understand the characteristics of internal genes of two strains of influenza A(H9N2) virns isolated from men and on the basis of these to reveal the origin of these two strains of influenza A(H9N2)virus. Methods The target gene was amplified by RT PCR,the PCR product was ligated with P GEM T Vector (Promega Company,U S A) at 4℃, the recombined plasmid was transferred into dH5α bacteria, and the positive colonies were selected and identified with restriction enzyme Afterwards, they were sent to Liu He Tong Company in Beijing for nucleotide sequencing Finally, phylogenetic analysis of the seguencing data was performed with MegAlign (version 1 03)and Editseq (Version 3 69)softwares Results Internal genes of the two strains of H9N2 virus were G9 lineage There was a slight difference in nucleotide sequence in PA gene between the two strains, whereas another five gene segments were identical to each other Conclusion The genomes of the two strains of influenza A(H9N2)virus were G9 lineage They were transmitted to men separately from different avian sources with different characteristics of gene of influenza A(H9N2) virus, respectively