牛病毒性腹泻病毒P20和P14基因的克隆及序列分析  

Cloning and sequence analysis of the P20 and P14 gene of bovine viral diarrhea virus

作　　者：李慧昕[1] 王君伟[1] 李宝臣[2] 韩先杰[1] 何海娟[1] 李昭春[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030 [2]黑龙江省生物制品一厂,黑龙江哈尔滨15000

出　　处：《中国预防兽医学报》2005年第2期124-129,共6页Chinese Journal of Preventive Veterinary Medicine

摘　　要：参考GenBank中BVDVOregonC2 4V株的基因组序列设计两对引物 ,利用套式PCR方法扩增P2 0基因 ,扩增出预期 5 2 5bp的目的片段。扩增产物克隆至pMD18_T载体 ,经酶切鉴定获得阳性重组质粒并对其进行测序。测序结果与参考序列OregonC2 4V比较 ,二者的核苷酸同源性仅为 80 95 % ,推导氨基酸同源性为 87 5 0 %。测序结果经NCBI上的Blast(Http :/www ncbi nlm nih gov/BLAST/ )同源性比较 ,克隆得到的基因与Osloss株同源性最高 ,核苷酸同源性为93 6 5 % ,推导氨基酸同源性为 95 83%。根据P2 0的测序结果 ,参照GenBank中BVDVOsloss株设计一对引物 ,扩增P14基因 ,经同源性比较 ,扩增的P14基因与Osloss株核苷酸同源性为 94 77% ,推导氨基酸同源性为 95 10 % ,通过系统发生分析 ,推测P2According to the sequence data of BVDV Oregon C24V strain published by GenBank,two set of primers were designed and used to amplify P20 gene by the method of nested polymerase chain reaction(PCR).A specific 525 bp DNA product was amplified,which was cloned into pMD18_T vector,and the positive recombinant clone was identified by restriction enzyme digestion.The recombinant plasmid was sequenced and compared with P20 gene of Oregon C24V strain.The nucleotide homololgy was 80.95 %,and the deduced amino acid homology was 87.50 %.Compared with blast on line (http://www.ncbi.nlm.nih.gov/bLAST/),the P20 gene of Osloss_like strain exhibits the highest homology with Osloss strain,they shares 93.65 % nucleotide sequence identity and 95.83 % amino acid identity.According to the sequencing result of P20 gene,one set of primers were designed for amplifying P14 gene on base of BVDV Osloss strain published by Genbank.The P14 gene of Osloss_like strain shares 94.77 % nucleotide sequence identity and 95.10 % deduced amino acid identity with Osloss strain.It suggests that P20 and P1 gene was phylogeneticly closer to BVDV Osloss strain after phylogenetic analysis.

关 键 词：牛病毒性腹泻病毒 P20基因和P14基因 克隆 序列分析 

分 类 号：S852.65[农业科学—基础兽医学] Q785[农业科学—兽医学]