6株猪O型口蹄疫病毒VP1基因的克隆与序列分析  

Cloning and sequence analysis of the VP1 gene of Guangdong derived strains of foot- and- mouth disease virus type O in swine

作　　者：娄高明[1] 张晋川[1] 张念祖[2] 

机构地区：[1]韶关学院英东生物工程学院,广东韶关512005 [2]云南省热带亚热带动物病毒病重点实验室,云南昆明650224

出　　处：《中国预防兽医学报》2006年第1期72-76,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：广东省科技攻关重大专项(№A204);韶关市重点科技攻关项目(韶科教2004-01);香港铭源基金重点项目(韶院科2004-01)

摘　　要：根据口蹄疫病毒(FMDV)VP1基因的序列,设计并合成了2对用于扩增VP1基因的引物。从组织中提取总RNA,首先用P1、P2引物对6株猪O型口蹄疫病毒进行RT—PCR扩增,获得1 000bp的片段;再用P3、P4引物进行巢式PCR扩增,结果获得850 bp的片段。将850 bp的片段克隆到pMD18—T载体中,通过PCR鉴定,将阳性重组质粒进行测序并分析。结果发现6株FMDV的核苷酸同源性为80.2%～99.4%,其推导的氨基酸序列同源性为86.9%～99.5%;构建遗传发生树,发现6株FMDV属于两个不同的基因型,其中的Shunde00、Sihui01、Shenzhen99、Fushan01株属一个基因型(与Hongkong93、广东86分离株属同一基因型);Guangzhou99、Shenzhen00株属另一个基因型(与UKG—12—2001株、JPN2000株属同一基因型)。通过对口蹄疫病毒VP1基因的测序与分析,了解其变异情况,为科学地防控FMD提供分子水平的依据。Two pair of primers amplified the VP1 gene of foot-and-mouth disease virus （FMDV） were designed and synthesized. The nucleotide sequences of VP1 gene of six strains named Shunde00,SihuiO1 ,Shenzhen99, FushanO1, Guangzhou99 and Shenzhen00 of FMDV type O were determined. RNA of the six strains of FMDV were extracted by single-step RNA isolation, reverse-transcripted and amplified by PCR, respectively. PCR products were cloned and sequenced. The homology among the six strains is 80.2% - 99.4% for nucleotide sequence and 86.9% - 99.5% for the deduced amino acid sequence. The nucleotide sequences of the six virus strains belong to two genetype. The ShundeOO,Sihui01,Shenzhen99 and FushanO1 strains share the same genetype with Guangdong86 strain and Hongkong93 strain; the Guangzhou99 and Shenzhen00 strains share the same genetype with UKG-12-2001 strain and JPN2000 strain. These results might be contributive to the selection of vaccine strains and analysis in molecular epidemiology.

关 键 词：口蹄疫病毒 VP1基因 遗传发生树 克隆 序列分析 

分 类 号：S852.6[农业科学—基础兽医学]