摘要
采用反转录－聚合酶链反应（ＲＴ－ＰＣＲ）方法，以自行设计的Ｈ７和Н５亚型禽流感病毒血凝素（ＨＡ）基因特异的两对引物，分别扩增了禽流感病毒Ａ／ＡｆｒｉｃａｎＳｔａｒｌｉｎｇ／９８３／７９（Ｈ７Ｎ１）株和Ｇ株（广东鹅体分离株，Ｈ５Ｎ１）约１．７ｋｂ的ＨＡ全基因ｃＤＮＡ。将所扩增的两个基因ｃＤＮＡ未端经Ｔ４ＤＮＡ聚合酶修饰后分别插入ｐＵＣ１８和ｐＢｌｕｅｓｃｒｉｐｔ质粒中，得到了两个基因的重组质粒。
Fowl plague (e i highly pathogenic avian influenza)is a severe infectious disease of chickens caused by highly pathogenic avian influenza virus (HPAIV) It is found that almost all isolates of HPAZVs beleng to H 5 and H 7 subtypes since 1959.Recently,H 5 HPAIV (G1 strain)was isolated from goose in China In the present study,two pairs of RT PCR primers specific to heamagglutinin(HA)genes of H5 and H7 subtypes respectively were designed based on the cDNA sequences of avian influenza virus (AIV) strains A/Chicken/Scotland/59(H5N1)and A/Chicken/Breslin/1902(H7N7) After that,two 1 7kb cDNA fragments were amplified respectively from two AIV strains G1 (H5N1)and A/African Starling/983/79(H7N1) The HA cDNAs of the AIVs of H5 and H7 subtypes were confirmed by restriction analysis Finally,two recombinant plasmids were constructed by inserting the H5 cDNA into pBluescript and the H7 cDNA into pUC18
