一株H9N2型禽流感病毒全基因的序列分析

采用异硫氰酸胍法提取禽流感病毒分离株的SPF鸡胚尿囊毒RNA,利用反转录-聚合酶链式反应(RT-PCR)技术扩增出禽流感病毒A/chicken/Zhuhai/154/03(H9N2)的8个基因片段(HA,NA,M,NP,NS,PA,PB1,PB2)。然后将其克隆到线状克隆载体pGEM-Teasy vector后进行序列测定和拼接,并将克隆到的8个基因片段与其他禽流感病毒毒株(A/Chicken/Guangdong/SS/94,A/Swine/Hong Kong/9/98-MA,A/Chicken/Shanghai/F/98,A/Duck/Hong Kong/Y439/97,A/Quail/Hong Kong/G1/97,A/Chicken/Shanghai/4/01)各个基因的相应序列进行比较分析。结果表明:克隆到的A/chicken/Zhuhai/154/03(AZH154)株的8个基因片段均含有相应病毒基因的完整开放阅读框架:AZH154的各基因与A/Chicken/Guangdong/SS/94株相应各基因同源性最高(NS基因除外,同源性只有67.5%),与A/Chicken/Shanghai/4/01各基因同源性高于A/Swine/HongKong/9/98-MA,A/Duck/Hong Kong/Y439/97和A/Chicken/Shanghai/F/98各基因同源性。所克隆的基因为禽流感病毒HA基因芯片的研究奠定了基础,同时为诊断和预防禽流感(AI)提供了科学依据。

The Analysis of the Entire Gene Sequence of H9N2 Avian Influenza Virus

In this experiment,the avian influenza isolate(H9)was propagated in SPF chicken embryo.Virus RNA was extracted from allantoic fluid by means of guandidinium isothiocyanate.The eight full-length cDNAs of an Avian Influenza Virus(AIV)A/chicken/Zhuhai/154/03(H9N2)was amplified by using RT-PCR.The cDNAs were then cloned to pGEM-T easy vector and sequenced.These sequences were compared with A/Chicken/Guangdong/SS/94,A/Swine/Hong Kong/9/98-MA,A/Chicken/Shanghai/F/98,A/Duck/Hong Kong/Y439/97,A/Quail/Hong Kong/G1/97 and A/Chicken/Shanghai/4/01.The results showed that each of eight gene cDNAs fragments contained the whole open reading frame.Most genes had high homology with A/Chicken/Guangdong/SS/94.The gene products provided a good foundation for the study of gene chips of AIV and for the diagnosis and prophylaxis of AI.