猪瘟病毒(HCV)石门株部分p23和p14基因的克隆、序列测定及与其他HCV株的比较

Sequence Analysis and Comparison of Part of p23 and p14 Genes in Hog Cholera Virus Isolates Shimen

应用ＲＴ－ＰＣＲ扩增猪瘟病毒（ＨＣＶ）石门株部分ｐ２３和ｐ１４基因，然后采用平端克隆法将扩增的ｃＤ－ＮＡ克隆到ｐＵＣ１９的ＳｍａⅠ位点，用双脱氧终止法对重组质粒中的插入片段进行序列分析。将所测定的序列与其他１０株ＨＣＶ相同基因区域用ＤＮＡＳＩＳ计算机软件进行同源性比较和系统树分析，结果表明，大部分毒株为同一个型，该型共包括９个ＨＣＶ株，以Ｂｒｅｓｃｉａ株为代表，石门（Ｓｈｉｍｅｎ）株也属该型；而Ａｌｆｏｒｔ株和Ｐ９７株与上述９个毒株差异较大，不属同一型。

One pair of oligonucleotide primers were designed to specifically amplify the region of the hog cholera virus (HCV) genome, which corresponds to a 629 bp portion encoding part of both p23 and p14. The product was amplified from HCV strain Shimen, the Chinese virulent reference strain. The cDNA product was cloned into pUC19 vector. Nucleotide sequencing was performed by Sanger′s method. Comparison and phylogenetic analysis of the nucleotide sequence of HCV strain Shimen and other 10 HCV reference strains was conducted by computer with DNASIS software. The results showed Shimen, Brescia and other 7 HCV strains should belong to the same genotype, but there was bigger difference between Alfort or P97 and other 9 HCV strains.