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口蹄疫病毒全长 cDNA 克隆的快速构建 

方先珍1， 2， 刘光清1， 刘在新1， 李平花1， 谢庆阁1

(1. 中国农业科学院 兰州兽医研究所， 甘肃 兰州 730046； 2.甘肃农业大学 动物医学院， 甘肃 兰州 730070)

摘要： 利用RTPCR和3′ cDNA末端快速扩增(RACE)技术分别扩增出包含口蹄疫病毒(FMDV)全基因组的2个重叠cDNA片段，将其分别插入到载体pcDNA3.1zeo(+)和pGEMTEasy中，然后利用片段本身单一酶切位点将FMDV全长cDNA克隆至pcDNA3.1zeo（+）载体，经PCR扩增、酶切鉴定和序列测定。结果表明，重组质粒（pcDNA3.1/FMDV）携带了FMDV OH/99基因组全长cDNA序列。

关键词： 口蹄疫病毒； cDNA末端快速扩增技术; 全长cDNA

中图分类号：S 852.659.6  文献标识码： A  文章编号：1000-6419(2004)05-0007-04

Rapid construction of fulllength cDNA clone of footandmouth disease virus

FANG Xian-zhen1,2, LIU Guang-qing1, LIU Zai-xin1, LI Ping-hua1, XIE Qing-ge1
(1. Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences,Lanzhou 730046,China; 
2. College of Veterinary Medicine,Gansu Agricutural University, Lanzhou 730070,China ) 

Abstract: Two overlap gene fragments of footandmouth disease virus(FMDV) were amplified by the reverse transcription polymerase chain reaction (RTPCR) and the 3’ rapid amplification of cDNA ends (3’ RACE) meathod. One was inserted into pcDNA3.1zeo(+) and the other was inserted into pGEMT Easy vector with single enzyme site .The recombinant plasmid pcDNA3.1/FMDV was identified by PCR, restriction analysis and DNA sequencing .The result showed that the recombinant plasmid pcDNA3.1/FMDV contains an FMDV fulllength cDNA.
Key words: footandmouth disease virus; rapid amplification of cDNA ends; fulllength cDNA

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