A组人轮状病毒DQ-1株VP7基因序列和蛋白质结构分析  

VP7 gene sequence and protein structure of human group A rotavirus strain DQ-1

作　　者：陈小萍[1] 林华[2] 柯益斌[1] 

机构地区：[1]福建省莆田学院医学院基础部生化教研室,莆田351100 [2]福建省莆田学院附属医院检验科

出　　处：《山西医科大学学报》2009年第10期877-881,共5页Journal of Shanxi Medical University

摘　　要：目的研究DQ-1株VP7基因的基因分型,对DQ-1株VP7基因序列和糖蛋白VP7的蛋白质结构进行分析。方法通过RT-PCR技术扩增人A组轮状病毒DQ-1株VP7全长基因,并将RT-PCR产物连接到pMD18-T载体,转化大肠杆菌JM109感受态细胞,在含有氨苄青霉素的LB琼脂培养基上筛选出两个PCR阳性菌落,并提取质粒,双酶切鉴定,最后利用通用引物进行测序。结果DQ-1株VP7基因与同型的我国已测序G1型地方株T73、R96-197株基因序列同源性均为97.6%,与G1型标准株Wa株核苷酸序列同源性为92.9%,与G2血清型代表株DS-1株基因序列同源性为73.8%,与G3血清型代表株SA11株核甘酸序列同源性为75.6%;其VP7基因mRNA可折叠多达20个发卡结构。DQ-1株氨基酸序列与T73、R96-197株同源性都达97.5%,与Wa株达94.6%,而与其他型DS-1株、SA11株和J-4621株分别仅为74.4%,79.4%和75.2%。结论DQ-1株属于G1血清型。DQ-1株VP7基因序列与已测序我国G1型地方株VP7基因序列几乎没有差别,而与G1型标准株VP7基因序列存在细微差异。Objective To study the genotype of strain DQ-1 VP7 gene,and to analyze DQ-1 VP7 gene sequence and protein structure. Methods Total RNA of DQ-1 that encoded the structural protein VP7 gene of GARV was amplified by reverse transcription polymerase chain reaction（RT-PCR）. Then the products of RT-PCR were ligated with plasmid pMD18-T and transformed into E. coli competent cells JM109. The VP7 gene of GARV strain DQ-1 was then sequenced. Results The nucleotide sequence of DQ-1 VP7 gene showed high identities to that of other human GARV such as T73 （ 97.6% ）, R96 - 197 （97.6%） and Wa（92.9% ） which belongs to GI genotype,while the identities to that of G2 DS-1 genotype and SA11 of G3 genotype were lower than 80%. The secondary structure of DQ-1 VP7 mRNA was constructed by about 20 stem-loop structures. GARV DQ-1 VP7 demonstrated 97.5% and 94.6% of amino acids similarity with I73 and Wa respectively, but only 74.4% ,79.4% and 75.2% with DS-1, SA11 and J-4621. Conclusion Strain DQ- 1 belongs to G1 serotype. VP7 gene of DQ-1 shares higher genotype identities with field strains of G1 serotype, but shows difference with standard strain.

关 键 词：轮状病毒 VP7结构蛋白 基因序列 蛋白质结构 

分 类 号：R512.5[医药卫生—内科学]