摘要
[目的]测定从仓鼠亚科宿主动物大仓鼠鼠肺中分离到的汉坦病毒的全S、部分M基因片段的核酸序列,明确其型别。[方法]现场采集鼠肺标本,IFAT筛选阳性鼠肺,分离病毒,用RT-PCR方法扩增,连接入PMD-18T载体,转化至JM109宿主菌。阳性质粒进行测序。[结果]YU62株S基因片段全长1701nt,有一个完整的ORF,起始位置为37nt,终止于1326nt,编码N蛋白429个氨基酸。与HTN-A16株核苷酸同源性最高为99.1%。M片段克隆出1272bp,核苷酸与HTN-SN7有85.6%的同源性。从S片段种系发生树分析来看,YU62归于HTN型H4亚型。从M片段(265￣624bp)种系发生树来看YU62株属于一个HTN的新亚型。[结论]仓鼠携带的汉坦病毒S、M片段进化不平行,YU62可能是在大仓鼠体内发生重排的HTN的新亚型。
[ Objective] TO set out the sequence of the nucleotide of complete S and partial M segment of the Hantavirus isolated from the lungs of cricetidae triton, and to categorize its genotype. [Methods] Firstly the segments were enlarged by RT-PCR, then linked to PMD-18T vector, and then transformed into JM109 host bacteria, the positive cloning plasmid was sequenced at last. [Results] The length of the complete nucleotide sequence of the S segment of YU62 strains was 1 701 nt. It contained one complete ORF encoding 429 amino acid of the N protein, which started at the 37th nt and ended at the 1 326th nt. The homology of nucleotide with HTN-A16 were 99.1%. M segment obtained 1 272 bp, 85.6% homology with HTN-SN7 was discovered. S segment homological analysis showed that YU62 belonged to H4 subtype of HTN genotype, M segment phylogenetic analysis suggested that YU62 was a new subtype of HTN. [Conclusion] S and M segment of YU62 evolve imparallelly,
出处
《现代预防医学》
CAS
北大核心
2007年第11期2076-2078,共3页
Modern Preventive Medicine
关键词
汉坦病毒
克隆
大仓鼠
Hantavirus
Cloning
Cricetidae triton
