【摘要】 设计了 Ｃ Ａ５、 Ｃ Ａ６ 和 Ｃ Ａ７、 Ｃ Ａ８ 两对引物，分别扩增鸡贫血病毒（ Ｃ Ａ Ｖ）山东分离株 Ｓ Ｊ１ 的 １５ Ｋｂ 和 ０８ Ｋｂ Ｄ Ｎ Ａ 片段。并将这两个片段分别克隆至 ｐ Ｕ Ｃ１１９ 和ｐ Ｂｌｕｅｓｃｒｉｐｔ（ Ｓ Ｋ＋ ）载体质粒，最后一起克隆到 ｐ Ｑ Ｅ３２ 载体质粒上，使两个片段前后连接成一个全长的病毒 Ｄ Ｎ Ａ。用该质粒转染 Ｍ Ｄ Ｃ Ｃ Ｍ Ｓ Ｂ１ 细胞，经免疫荧光抗体法和 Ｅ Ｌ Ｉ Ｓ Ａ 检测到 Ｃ Ａ Ｖ 病毒，结果证明我们得到了一个 Ｃ Ａ Ｖ 感染性全基因克隆。更多还原

【Abstract】 The 1.5Kb fragment and the rest 0.8kb fragment of CAV isolate SJ1 from Shandong were amplified by two pairs of primers,CA5 CA6 and CA7 CA8,both of which contain EcoRI and BamHI sites respectively.The two fragments were cloned into EcoRI BamHI site of plasmid pQE32 and plasmid pBluescript(sk+),respectively.The 0.8kb fragment was then transrerred from pBluescript(sk+)to the former pQE32 by BamHI/KpnI digestion and subsequent ligation,to form plasmid as CAV pQE32 which contained the whole genome of CAV linearized at EcoRI sites.The CAV pQE32 was proved to be an infectious clone by transfection of MDCC MSB1 cell line The produced CAV virus detected both by immunofluence and ELISA.更多还原