甲_1型流感病毒新分离株HA基因的序列分析

Gene analysis of new isolates HA of influenza A (H_1N_1) virus

目的研究新分离的 H1 N1 亚型流感病毒株的 HA1基因序列。方法甲型流感病毒通过鸡胚增殖后提取 RNA、逆转录合成 c DNA,经 PCR扩增和产物纯化构建重组质粒 ,用双脱氧链终止法进行核苷酸序列测定并进行基因特性分析。结果新分离到的 3株流感病毒株 (H1 N1 ) HA 1区基因长度为 981bp,编码 32 7个氨基酸 ;与 A/桂防 / 10 / 94和 A/ Bayern/ 0 7/ 95(H1 N1 )标准株比较其同源性分别为 92 .8%和 91.3% ,丢失了第 130位氨基酸和 30 4位糖基化位点 ;新分离的 3株甲型流感病毒株 (H1 N1 ) HA1区氨基酸同源性高达 98% ;A/桂防 / 10 / 94和 A/ Bayern/ 0 7/ 95 (H1 N1 )毒株 HN1氨基酸的同源性高达 96 %。结论新分离到的 3株 H1 N1 毒株 HA编码氨基酸不同于 A/ Bayern/ 0 7/ 95 (H1 N1 )和 A /桂防 / 10 / 94(H1 N1 )标准株 。

ObjectiveTo obtain genetic characterization of the HA1 of new isolates of influenza A (H 1N 1) virus MethodsVirus was amplified in embryonated chicken eggs, the virion DNA was transcribed into cDNA by reverse transcriptase, cDNA amplified by PCR, the products of PCR were purified Afterward, DNA sequence analysis was performed by the dideoxy-mediated chain termination method, using synthetic oligo nucleotide primers ResultsThe HA1 domain of new isolates of influenza A (H 1N 1) virus showed that their HA1 genes were 981 nucleotide in length coding for a HA1 protein with 327 amino acids, deletion of a glycosylation site and an amino acid The homology of amino acid sequences of protein molecules on HA1 domains of new influenza A virus when compared with A/Guifang/10/94(H 1N 1) and A/Bayern /07/95 (H 1N 1) viruses,was 92 8% and 91 3% respectively; The homology of amino acid sequences of protein molecules on HA1 domains of A/Jingjun/11/98(H 1N 1) when compared with A/Jingjun/13/98(H 1N 1) or A/Jingjun/807/97(H 1N 1) viruses,was as high as 98%; The homology of amino acid sequences of protein molecules on HA1 domains of A/Guifang/10/94(H 1N 1) when compared with A/Bayern/07/95(H 1N 1) viruses was as high as 96% ConclusionsThe HA1 gene of new H 1N 1 virus strains is different from those of A/Guifang/10/94 (H 1N 1) and A/Bayern/07/95(H 1N 1), they will probably be new mutation strains