呼吸道合胞病毒G蛋白胞外区基因克隆及序列分析  

Molecular Cloning and Sequence Analysis of The Ectodomain of Protein G of A Local Strain Respiratory Syncytial Virus Isolated from Guangzhou Area

作　　者：钟南山[1] 黄海鹭[1] 常汝虚 马炳南 

机构地区：[1]广州呼吸病研究所,广州510120

出　　处：《现代临床医学生物工程学杂志》2000年第3期163-167,共5页Journal of Modern Clinical Medical Bioengineering

基　　金：国家自然科学基金资助项目!(批准号39870325);广东省自然科学基金资助项目

摘　　要：目的　对呼吸道合胞病毒G蛋白胞外区基因进行克隆及序列分析 .方法　从RSV感染的喘息型支气管炎患儿咽试子分离的病毒培养物中提取病毒RNA ,进行逆转录及PCR扩增 ,克隆到G蛋白胞外区基因DNA序列分析 .结果　克隆片段全长 6 90bp ,编码一个含有 2 2 9个氨基酸的多肽 .与 5个GenBank中不同的RSV分离株(包括A ,B亚型 )的G蛋白胞外区的氨基酸序列相比 ,A亚型的RSV毒株氨基酸水平的同源率为 95 .2 %～86 .0 % ,B亚型的氨基酸水平同源率为 44 .3% .氨基酸的变异主要集中在胞外区一个高度保守的两端 .该G蛋白胞外区的核苷酸序列与A亚型RSV -G32 3相比同源率为 97.9% .结论　广州地区RSV分离株 (9815 9)属于A亚型 ;其G蛋白胞外区基因具有较高的变异性 。Objective The nucleotide sequence of the ectodomain of glycoprotein (G) from a local strain respiratory syncytial virus in Guangzhou area was determined.Methods The viral RNA was extracted from sample of the throat swab in bronchitic children with RSV infection. The complementary DNA, which encodes ectodomain of gene, was synthesized and amplified by polymerase chain reaction (PCR).The product was cloned into Pbluescript and the complete nucleotide sequence was determined.Results There is an open reading frame of 690 nucleotides which encode 299 amino acids of the glycoprotein ectodomain. It shares 95.2%～86.0% amino acid sequence homology with five A-subgroup strains and one B-subgroup strain, respectively. The amino acid variations are mainly located in the extracellular part of the protein where two extensive divergent domains with a highly conservative region existed in between.Conclusions The sequence analysis indicated that the RSV isolate (G98159) is an A-subgroup strain. This study demonstrates the sequence diversity of the glycoprotein ectodomain of RSV between a Guangzhou isolate and others published RSV isolates.

关 键 词：呼吸道合胞病毒 基因克隆 序列分析 RSV-G蛋白 

分 类 号：R373[医药卫生—病原生物学]