摘要
利用 PCR技术 ,从纯化番鸭细小病毒 M91 G2 7毒株鹅胚尿囊液中扩增出病毒结构多肽 VP3基因。将该 PCR扩增片段克隆入 p UC1 8质粒载体的 H inc 和 Sac 位点之间 ,酶切分析筛选到含 1 .6kb基因片段的重组质粒 MP1 3。结果表明 :该片段与国外已报道毒株核苷酸序列有 98.8%的同源性 ,氨基酸序列有 98.1 %的同源性 ,证明该重组质粒是 VP3基因的克隆。
Using polymerase chain reaction (PCR) technique, VP 3 gene was amplified from the DNA of one field isolate of Muscovy parvovirus (M 91 G 27 ) isolated in China. The PCR amplified VP 3 gene was cloned into the HincⅡ and SacⅠ sites of pUC18. Restriction endonuclease analysis showed that the recombinant plasmid MP 13 contained 1.6 kb fragment was obtained. Sequence and analysis showed that the Chniese isolate shared 98.8% and 98.1% identity with the sequence repor ted by foreign scholar at nucleotide and amino acid levels respectively, and indicted that the recombinant plasmid MP 13 contained the VP 3 gene.
