番鸭细小病毒MDPV-Q株VP2蛋白基因的克隆与序列测定

Cloning and Sequencing of DNA Encoding with VP2 Structural Protein of Muscovy Duck Parvovirus

根据genebank中番鸭细小病毒 (MDPV)的基因序列 ,设计了一对引物LHMP7/LHMP8,同时在这 2条引物中分别加入 2种限制性核酸内切酶SacII和KpnI的酶切位点 ,使扩增后的DNA片段的两端 ,分别含有这 2种酶的酶切位点。应用PCR技术扩增了MDPV_Q株的VP2蛋白基因片段。将扩增后的VP2蛋白基因重组到pMD18_T质粒载体上 ,并对插入片段进行序列测定。测序结果表明 ,我国分离的MDPV_Q株基VP2蛋白基因序列与国外分离株的同源性达 97.9%。

According to complete nucleotide sequences of Muscovy duck parvovirus registered in the gene bank,two modified primers(LHMP7/LHMP8)were designed and,for each of them,a restriction endonuclease recognition site,SacⅡ or KpnⅠ,was included respectively,in order to the amplified DNA fragment contains a restriction endonuclease recognition site at its ends after PCR amplification.The DNA encoding VP2 structural protein was amplified by PCR,recombined into pMD18_T vector and sequenced.It shows that the homology of both DNAs encoding VP2 structural protein of MDPV_Q and MDPV which were registered in the gene bank is 97.9%.