广西猪瘟流行毒株全基因组的克隆与序列分析  

Cloning and Sequence Analysis of Genome of Classical Swine Fever Virus GXWZ02 Strain Epidemic in Guangxi

作　　者：吴显实[1] 罗廷荣[1] 廖素环[1] 刘芳[1] 陆芹章[1] 黄伟坚[1] 温荣辉[1] 秦爱珍[1] 余克伦[1] 

机构地区：[1]广西大学动物科学技术学院,广西南宁530005

出　　处：《中国兽医学报》2005年第2期125-128,共4页Chinese Journal of Veterinary Science

基　　金：广西科技厅科研基金项目(桂科青9912015)

摘　　要：参考已发表的猪瘟病毒 (CSFV) Shimen株、HCL V株、Paderborn株的核苷酸序列 ,设计并合成 1 1对引物 ,应用RT- PCR技术 ,成功地分 1 1个片段扩增了 CSFV广西流行毒株 GXWZ0 2的全基因组。将这 1 1个基因片段克隆 ,并测定了其核苷酸序列。应用计算机生物软件 Vector NTI将 1 1个基因片段进行拼接 ,确认 CSFV GXWZ0 2株全基因组序列的长度为 1 2 2 96个核苷酸 (Gen Bank收录号 AY3 6 776 7)。将 GXWZ0 2株与国内外已发表的 Shimen、HCL V、3 9、Brescia、Eystrup、Glentorf、Alfort1 87、Paderborn、CS、AL D、GPE- 、P97、L PC毒株进行全基因组核苷酸序列和推定的氨基酸序列比较 ,核苷酸同源性分别为 85 .4 %、 84 .6 %、88.7%、85 .7%、 85 .7%、 85 .3 %、85 .6 %、 95 .3 %、 85 .7%、85 .7%85 .4 %、83 .4 %、84 .3 % ,推定的氨基酸同源性分别为 92 .6 %、91 .7%、94 .2 %、 93 .1 %、92 .9%、92 .3 %、 93 .0 %、97.7%、92 .6 %、93 .0 %、92 .6 %、90 .5 %、90 .6 %。 GXWZ0 2与 Shimen、HCL V的全基因组序列及推定的氨基酸序列有明显差异 ,表明 GXWZ0 2株在遗传性和抗原性上发生了较明显的变化。系统发育树分析 ,所比较的 1 4株 CSFV被分为2个群 :GXWZ0 2、Paderborn和 3 9这 3个毒株被归为群 .In this study,11 pairs of specific primers were designed and synthesized.Using these primers,Eleven fragments had been successfully amplified from CSFV GXWZ02 strain by RT-PCR.These amplified fragments were cloned and sequenced.Assembly of sequences of the 11 fragments into a contiguous sequence was done using Vector NTI software.There are (12 296) nucleotides in the genome of GXWZ02 strain.Comparison of the genomic nucleotide sequence and the deduced amino acid sequence of GXWZ02 with that of Shimen,HCLV,39,Brescia,Eystrup,Glentorf,Alfort187,Paderborn,CS,ALD,GPE^(-),P97 and LPC,showed that the nucleotide homologies were 85.4%,84.6%,88.7%,85.7%,85.7%,85.3%,85.6%,95.3%,85.7%,85.7%,85.4%,83.4% and 84.3%,respectively,the amino acid sequence homologies were 92.6%,91.7%,94.2%,93.1%,92.9%,92.3%,93.0%,97.7%,92.6%,93.0%,92.6%,90.5% and 90.6%,respectively.The homologies of genomic nucleotide sequence and deduced amino acid sequence of CSFV GXWZ02 with that of Shimen,HCLV were low,indicating that GXWZ02 was genetically different from Shimen and HCLV.Phylogenetic tree analysis based on the genomic nucleotide sequence showed that the 14 CSFV stains belonged to two groups.GXWZ02,Paderborn and 39 were classified into groupⅠ,and the other strains were classified into groupⅡ.The genetic relationship is very close between GXWZ02 and Paderborn.Comparison of the nucleotide sequences and the deduced amino acid sequences of different gene of GXWZ02 with that of Shimen,HCLV,demonstrated that the genetic variation rate of E^(rns),E2,P7,NS2 and NS5A genes was high,while that of NS3,NS4A and NS4B genes was low.

关 键 词：猪瘟病毒 基因组克隆 序列分析 GXWZ02株 猪瘟 

分 类 号：S852.659.6[农业科学—基础兽医学]