摘要
用RT PCR法 ,以口蹄疫病毒 (FMDV )WH/ 99株牛舌水泡皮为材料 ,提取RNA及扩增目的cDNA ,然后与pGEM TEasy载体连接并转化JM 10 9菌株 ,再经重组质粒电泳、PCR和EcoRⅠ酶切鉴定 ;用DNAstar软件比较功能未知区的序列差异 ,以RNAdraw软件分析它们的二级结构。结果表明 ,7株FMDV的功能未知区序列长度介于 2 0 7— 2 5 6个核苷酸 ;序列分析表明 ,WH/ 99与AsiaⅠ 63 / 72、A2 4 /Cruzeiro、TWTY/ 97和TWCP/ 97的序列差异最大 ,预示此区段可能与病毒感染嗜性有关 ;二级结构分析表明 ,A2 4 /Cruzeiro、AsiaⅠ 63 / 72和TWCP/ 97均有 5个结构域 ,WH/ 99和TWTY/ 97具有 6个结构域。TWTY/ 97和TWCP/ 97二级结构的差异可能是TWTY/ 97功能未知区第 2 0位插入的“C”所致 ,而O1K/66和O1Campos仅有 3个结构域。 2株猪O型FMDV功能未知区 5′端存在着连续的核苷酸缺失现象 ,最多达 5 2个 ,唯有3株牛O型FMDV(O1K/ 66、O1Campos和WH/ 99)在上述位置无任何缺失。
The sequence of function-unknown region b et ween poly(C) and IRES tract in foot-and-mouth disease virus strain WH/99 from bovine tongue epithelium was determined by cloning of cDNA fragments ob tained from RT-PCR. The DNA fragments were inserted in pGEM-T vectors and transformed into JM109, then the reco mbinant plasmids were identified by electrophoresis, EcoRⅠ cleavage and PCR. T he nucleotide sequence of the function-unknown region was compared with six oth er reference strains and the secondary structures of all these strains were drow n by the software RNAdraw. The sequence alignment shown that the length of the f unction-unknown region is between 207—256 nucleotides, and sequence differentia of WH/99 with AsiaⅠ63/72, A_(24)/Cruzeiro and Taiwan strains is more obviou s than other strains, the region might related with virus infection for animals. In the secondary structure, there were Five domains in A_(24)/Cruzeiro, Asia Ⅰ63/72 and TWCP/97; three domains in O_(1)K and O_(1)Campos; six domains in WH/99 and TWTY/97. The continuous nucleotides were deleted at the 5′end of t he function-unknown region of FMDV isolated from infected swine, height to 52 nu cleotides, there were no any deletions at same position in three FMDV strains ty pe O from infected bovine.
出处
《中国兽医科技》
CSCD
北大核心
2001年第11期5-9,共5页
Chinese Journal of Veterinary Science and Technology
基金
国家"973"项目 :畜禽重大疫病病原大分子结构与功能研究 (G1 9990 1 1 90 1 )部分内容
