汉坦病毒中国疫苗株Z37M片段的克隆及序列分析

Molecular Cloning and Nucleotide Sequence Analysis of the M Genome Segment of Hantavirus Z37, a Chinese HFRS Vaccine Strain

汉坦病毒Z37株是从褐家鼠体内分离到的 ,用于生产双价肾综合征出血热疫苗的病毒毒株之一 ,血清分型为SEO型。利用RT PCR方法扩增Z37株M基因片段cDNA ,克隆入质粒载体 ,进行核苷酸序列测定及分析。Z37株M基因片段由 36 5 1个核苷酸组成 ,只有一个开放读码框架 ,共编码 1133个氨基酸。与HTN型病毒 ( 76 118、A9、HV 114)的核苷酸和氨基酸同源性分别为 71.8%～ 72 .1%、76 .2 %～ 76 .7% ,与SEO型 (R2 2、L99、80 39)的核苷酸和氨基酸同源性分别为 95 .3%～ 96 .1%、95 .3 %～ 98.9%。这一结果的获得进一步从分子水平确定了Z37株的型别 。

One of the strain of bivalent HFRS vaccine, Z37 strain was isolated from Rattus norvegicus and identified as SEO virus by serological test. The M segment cDNA of Hantavirus Z37 strain was obtained by reverse transcription and polymerase chain reaction, subsequently cloned into pGEM T vector. The sequence of positive recombinants was determined by the method of dideoxy chain termination, which revealed that the M genomic segment is 3651 nucleotide in length with a predicated long open reading frame encoding a protein of 1133 amino acids. Comparison with HNT type (76 118, A9, HV 114 strains) indicated that there were 71.8%～72.1% homology at the nucleotide level, 76.2～76.7% homology at the amino acid level. Comparison with SEO type (R22,L99,80 39 strains) showed 95.3～96.1 homology at the nucleotide level, 95.3～99.2% homology at the amino acid level. The results of nucleotide and amino acid comparison indicated that Z37 strain is SEO viruses in molecular level.