摘要
采用RT-PCR方法对FMDV OH99株基因组全序列进行了分子克隆与测序。结果表明OH99株基因组全基因组序列长8040nt,其中5’NCR长1026nt,前导蛋白(L)编码区长603nt。该毒株结构蛋白与非结构蛋白编码区的核苷酸序列为6318nt,3’NCR长93nt,其后是poly(A)尾巴,测序结果表明该结构至少含有56个A。应用分子生物学软件,将OH99株与其它参考毒株进行了序列比较,并对其基因特征、推导的氨基酸序列进行了研究分析。结果显示,在分类地位上OH99株归属于O型FMDV,与OTY TW/97具有较高的同源性,而与其他参考毒株的差异性比较大,而且在基因组功能未知区域和3A编码区域具有两处明显的基因片段缺失现象,其中3A编码区缺失30nt,与OTY TW/97株相同,但功能未知区域的缺失状况与OTY TW/97稍有差异。根据VP1基因序列,对OH99株与参考毒株进行了系统发生树分析,分析结果表明OH99株与OTY TW/97株在同一基因型内,其遗传关系最近,而与其毒株遗传关系较远。
The full-length genome of Foot-and-Mouth disease virus (FMDV) OH99strain was cloned by RT-PCR and 3'RACE, and sequenced. The result shows that the full-length genome of OH99 strain is 8040nt in length, including 5'NCR, Leader protein coding region, Polyprotein coding region, and 3'NCR Their nucleotide sequence is 1026nt, 603nt, 6318nt, and 93nt respectively. Besides, poly (A) tail was also coined by 3'RACE, it's 56nt in length. The full-length genome of OH99 strain was compared with other reference strains, and its genome characteristic is also analyzed. With the help of some biological ware, The Phylogenetic tree obtained from the nucleotide sequence of VPlgene from OH99 strain and ten reference strains shows that OH99 strain could be ranked FMDV O style. Furthermore, this strain and OTY TW/97 are in the same genotype, their nucleotide sequence share high homology, but it shows remarkable divergence with other reference strains, including china99 strain. For example, two short genome segment was deleted in the function-unknown region and 3A coding region of OTY TW/97 and OH99 strain. Which would affect the host tropism of virus. The results suggest OH99 and OTY TW/97 may come from the same ancestor, but the genetic distance between OH99 and other reference strains is far.
出处
《中国病毒学》
CSCD
2003年第3期259-264,共6页
Virologica Sinica
基金
国家"973"项目资助(G1999011901)