黑龙江省首次发现汉坦病毒家鼠型的分子生物学证据  

Molecular biological evidence of Seoul virus firstly isolated in the Northeast China (Heilongjiang province): Complete nucleotide sequence analysis of the S genome segment of hantavirus Pf26 strain

作　　者：孙成群[1] 陈露菲[1] 吴玉华[1] 陶伟英[1] 刘彦成[1] 崔岩[1] 许君[1] 李冀宏[1] 刘忠伟[1] 孙荣芳[1] 甄尚敏[1] 张秀彬[1] 

机构地区：[1]黑龙江省卫生防疫站病毒病防治研究所,黑龙江哈尔滨150036

出　　处：《疾病控制杂志》2000年第4期313-316,共4页Chinese Journal of Disease Control and Prevention

基　　金：国家计委资助的重点科技项目!黑龙江省灾区 HFRS防制的研究 (无编号 )

摘　　要：目的　测定从疫区捕获的宿主动物 -褐家鼠肺中分离的汉坦病毒 Pf2 6株的 S基因片段的全基因序列。方法　选择汉坦病毒 S基因序列保守区设计引物 ,用 RT- PCR方法将 S基因分两段扩增 ,再将扩增片段连接到 PU C19载体上 ,然后进行测序和分析。结果　 Pf2 6株的 S基因片段的全序列长为 1776 nt。只有一个主要的编码 N蛋白的 ORF,起始位置为第 43 nt,终止于1332 nt,编码的蛋白长为 42 9aa。 Pf2 6株的 S基因片段核苷酸序列与 SEO型中的病毒株的同源性最高 ,而与汉坦病毒其他型的同源性较差。从种系发生树分析来看 ,Pf2 6株归结于汉坦病毒的SEO型。在 SEO型之内 ,从 S基因的种系发生树 (A)上看 ,Pf2 6株与 sr- 11株的亲缘关系比较近。但是 ,从编码的 N蛋白的种系发生树 (B)上看 ,Pf2 6株与 Z37株、Guo3株的亲缘关系比较近。Pf2 6株的 S基因和 N蛋白序列与汉坦病毒 SEO型分离株 sr- 11,Z37,Guo3株的同源性最高 ,分别为 :核苷酸同源性 ,97% ,96 % ,88% ;氨基酸同源性 ,97% ,98% ,98%。Pf2 6株与 sr- 11株 ,Guo3株 ,Z37株 N蛋白氨基酸变异主要发生在 N蛋白的前半部分。中国分离到的 Pf2 6 ,Guo3,Z37株与 sr- 11株氨基酸变异主要集中在 aa6 9- aa75位之间。结论　 Pf2 6株为一株新的汉坦病毒 SEO型病毒株。Objective To determine the complete nucleotide sequence of the S genome segment of Pf26 virus strain isolated from a rat (Rattus norvegicus) in Heilongjiang province. Methods The complete nucleotide sequence of the S genome segment of Pf26 virus strain was determined with two clones, which were obtained by RT PCR,and by linking two overlapping fragments into PUC19 vector respectively. Results The complete nucleotide sequence of the S genome segment of Pf26 virus strain included 1 776 nucleotides in length and contained one ORF encoding the N protein started at the 43rd nt and ended at the 1 332nd nt. The length of encoded protein, 429 amino acids, is identical to the N protein of SEO type of hantaviruses. Comparison of S segment sequence of Pf26 strain with the corresponding sequences of other hantaviruses showed that the identities of Pf26 strain to sr 11, Z37 and Guo3 strains were 97%,96%,88% in complete nucleotide sequence of S genome segment and 97%,98%,98% in amino acid sequence of N protein respectively. Phylogenetic analysis showed that sr 11, Z37, Guo3 and pf26 strain formed one lineage within the clade containing hantaviruses of SEO type. Conclusions Pf26 strain belongs to hantavirus SEO type.

关 键 词：汉坦病毒 克隆聚合酶链反应 碱基序列 

分 类 号：R373.32[医药卫生—病原生物学] Q78[医药卫生—基础医学]