我国新城疫强毒株F糖蛋白基因的体外合成及其抗原性研究

In vitro synthesis of F protein gene from a Chinese virulent strain of NDV and its antigenicity to specific antibody probe

将我国新城疫强毒株Ｆ４８Ｅ９在ＳＰＦ鸡胚增殖后提取ＲＮＡ，用自行设计的一对寡聚核苷酸引物进行ＲＴ－ＰＣＲ，合成了Ｆ糖蛋白基因片段。将此片段与ｐＵＣ１９质粒重组，转化大肠杆菌ＪＭ１０９，从阳性克隆中提取重组质粒，再用ＰＣＲ法扩增，获得了同样大小的ＤＮＡ片段。经用特异性抗体探针检测。

A 1.9kb of F protein gene fragment was amplified from a virulent strain (F 48 E 9) of NDV isolated in China by RT PCR.Recombinants were obtained after the F protein gene fragment was inserted in pUC19 DNA between the Hind Ⅲ and Sal Ⅰ sites and transformed into E.coli JM109.In the identification trials,the same F protein gene fragment could be amplified by PCR and cleaved by Hind Ⅲ and Sal Ⅰ from the recombinant DNA.Using an improved method of colony hybridiztion in situ,the positive clones of E.coli JM109 appeared good antigenicity to the specific antibody probe.