猪繁殖与呼吸综合征病毒(PRRSV)BJ-4株全基因组序列测定与分析  

Sequencing of Genome of Porcine Reproductive and Respiratory Syndrome Virus isolate BJ-4

作　　者：杨汉春[1] 黄芳芳[1] 郭鑫[1] 高云[1] 李华[1] 陈声[1] 

机构地区：[1]中国农业大学动物医学院农业部预防兽医学重点开放实验室,北京100094

出　　处：《农业生物技术学报》2001年第3期212-218,共7页Journal of Agricultural Biotechnology

基　　金：国家重点基础研究发展规划项目"畜禽重大疫病病原大分子结构与功能研究(G1999011901)资助

摘　　要：在国内首次完成了猪繁殖与呼吸综合征病毒分离毒株的全基因组序列测定。用ＲＴ－ＰＣＲ方法分段扩增出ＰＲＲＳＶ北京地区分离株ＢＪ－４株的２１条ｃＤＮＡ片段，分别克隆于ｐＧＥＭ－Ｔ－ｅａｓｙ质粒载体并进行测序，按顺序将这些序列进行拼接得到ＰＲＲＳＶ　ＢＪ－４株全基因组ｃＤＮＡ序列。测序结果表明ＰＲＲＳＶ　ＢＪ－４株基因组全长１５　５０４　ｂ，包含８个开放阅读框，５′端有１９０　ｎｔ先导序列；３′端有２５６　ｎｔ　ＵＴＲ，其中包括９５　ｎｔ　Ｐｏｌｙ（Ａ）尾。ＰＲＲＳＶ　ＢＪ－４株与美洲型毒株ＶＲ－２３３２和Ｎｅｂｒｅｓｋａ的核苷酸同源率分别为９９．５％和９８．４％，而与欧洲型代表株ＬＶ株的核苷酸同源率为６２．５％，与另一国内分离株ＣＨ－１ａ株编码结构蛋白基因的核苷酸同源率为９３．３％。Complete nucleotide sequence of genome of porcine reproductive and respiratory syndrome virus(PRRSV)isolate was first reported in this paper in China. Twenty-one cDNA fragments of PRRSV BJ-4 genome was amplified using RT-PCR, and cloned into pGEM-T-easy plasmid vectors respectively and sequenced. Complete nucleotide sequence of genome of PRRSV BJ-4 was obtained by spicing sequence of each fragment in order. It showed that the genome of PRRSV BJ-4 was 15 504 b in length, containing eight open reading frames(ORFS)with 190 nt leader sequence in the 5'nd of the viral genome and 256 nt uncoding terminal region(UTR)in the 3'nd of the genome, including 95 nt poly(A)tail.Compared with the American isolate VR-2332 and Nebreska isolate, the genome of PRRSV BJ-4 showed 99.5% and 98.4% of nucleotide homology respectively. With European Lelystad Virus(LV),PRRSV BJ-4 only showed 62.5% nucleotide similarity. There was a 93.3% nucleotide homology in structural protein gene compared with the isolate CH-1a.

关 键 词：PRRS病毒 全基因组 序列测定 猪病毒 

分 类 号：S852.65[农业科学—基础兽医学]