【摘要】 根据国外已发表的胸膜肺炎放线杆菌 (APP) Apx 的基因序列设计了 1对引物 ,从我国 APP武汉株扩增出约3.5 kb的 apx CA基因。测序结果表明 ,该基因的大小与国外已报道的 1型和 9型 APP的 apx CA基因大小一致 ,同源性达 99%。将该基因连接到原核表达载体 p ET- 17b,构建了原核表达质粒 p ET- apx CA,并在大肠杆菌 BL 2 1(DE3)中表达出了大小约 10 5 0 0 0的 Apx 蛋白。提取包涵体蛋白作为抗原包被 EL ISA酶标板 ,对已知阳性、阴性血清和临床送检血清样品进行了 EL ISA检测 ,从而为最终建立 Apx - EL ISA检测方法奠定了基础更多还原

【Abstract】 The 3.5 kb toxic gene apxⅡCA of Actinobacillus pleuropneumoniae was got and amplified by PCR after a pair of particular primer was deviced according to the published sequence abroad.The sequencing result showed that the size of this gene was the same as that published in serotype 1 and 9,and it reached 99% homogeneity of the corresponding gene in serotype 1 and 9.The gene apxⅡCA was cloned into prokaryotic expression vector pET-17b to develop the recombinant plasmid pET-apxⅡCA,which was transformed into BL(DE3),as a result,a particular (105 000) protein was produced.The inclusion protein was exacted and used as immunogent to coat the polystyrene U-bottom microtiter plates to detect clinical sera.This study is a preliminary step for enzyme-linked immunosorbent assay for Actinobacillus pleuropneumoniae.更多还原