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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 ,该研究为RSV蛋白的免疫机理及RSV疫苗打下基础
Objective To clone and sequence the extracellular domain of respiratory syncytial virus G protein.Methods The viral RNA was extracted from the virus culture of pharyngitis in RSV-infected asthmatic bronchitis patients for reverse transcription and PCR amplification, And cloned into the extracellular domain of G protein.Results The cloned fragment was 690bp in length and encoded a polypeptide of 229 amino acids.Compared with 5 different RSV isolates (including A and B subtypes) in GenBank The amino acid sequence of the extracellular region of G protein was 95.2% ~ 86.0% of that of the RSV strain of subtype A, and 44.3% of the B subtype. The amino acid variation mainly concentrated on one highly conserved end of the extracellular domain.The nucleotide sequence of the extracellular domain of G protein was 97.9% compared with the RSV-G32 3 subtype A. Conclusion The RSV isolates (9815 9) belonged to A subtype. The G protein extracellular region gene had high variability. This study lays the foundation for the immunological mechanism of RSV protein and RSV vaccine