A组人轮状病毒VP3基因克隆及重组表达

发布时间：2018-01-07 18:09

本文关键词：A组人轮状病毒VP3基因克隆及重组表达　出处：《北京协和医学院》2011年硕士论文　论文类型：学位论文

更多相关文章： 轮状病毒 VP3 重组表达 基因分型

【摘要】：轮状病毒(rotavirus,RV)属于呼肠病毒科(Reoviridae)轮状病毒属(Rotavirus),是引起婴幼儿严重胃肠炎的主要病原体,每年全球因RV感染所导致的重症腹泻而死亡的儿童约600000。RV是直径约为70nm,无包膜,由3层衣壳蛋白组成的正二十面体双链RNA病毒。RV基因组包含11个分节段的双链RNA,编码6个结构蛋白(VP1-VP4, VP6, VP7)和6个非结构蛋白(NSP1-NSP6)。根据结构蛋白VP6的抗原性差异,可将已发现的RV分为A-G7个组,其中,A组为引起婴幼儿腹泻的主要病原。 RV结构蛋白VP3在病毒基因转录过程中发挥着重要的功能。本研究运用基因克隆和重组表达技术,构建了携带A组人RV TB-Chen株VP3蛋白完整编码基因的重组表达质粒,在大肠杆菌中表达重组VP3蛋白(rVP3),对A组RV VP3进行了系统进化和基因分型研究。结果表明,rVP3可在大肠杆菌BL21(DE3)细胞中表达；表达的完整rVP3在菌体中以包涵体形式存在；rVP3可以被抗SA1l株RV全病毒蛋白的豚鼠血清抗体识别；至今发现的A组RVVP3至少可以分为7个不同的基因型(M1-M7),其中人类中有3个型(M1-M3),M4和M7型仅在鸟类中发现,TB-Chen株的VP3基因属于M2型；不同VP3基因型有独特的敏感宿主范围,同一基因型可能感染不同种动物,同一种动物也可能感染不同基因型。研究结果为大量制备RV VP3蛋白及进一步研究其结构功能提供了重要基础。
[Abstract]:Rotavirus (rotavirus, RV) belonging to the Reoviridae (Reoviridae) rotavirus (Rotavirus), is the major pathogen causing severe gastroenteritis in infants, every year around the world because of the death of RV infection caused by severe diarrhea of children about 600000.RV is about 70nm in diameter, without capsule, twenty sided double chain RNA virus.RV genome consists of 3 layers of double stranded RNA capsid protein contains 11 segments, 6 structural proteins (VP1-VP4, VP6, VP7 encoding) and 6 non structural protein (NSP1-NSP6). According to the antigenic difference of structural protein VP6 of RV has been found, can be divided into A-G7 groups, including, group A was mainly caused by the pathogen of infantile diarrhea.
RV VP3 protein played an important role in viral gene transcription. In this study the use of gene cloning and recombinant expression technology, construct the intact VP3 protein encoding gene of human group A RV TB-Chen strain of recombinant expression plasmid, recombinant VP3 protein expressed in Escherichia coli (rVP3), the A group VP3 was RV study on system evolution and gene. The results showed that rVP3 in Escherichia coli BL21 (DE3) cells; complete expression of rVP3 in bacteria in the form of inclusion body; guinea pig serum rVP3 antibody recognition can be anti SA1l strains of RV virus protein; A RVVP3 has found the group can be divided into at least 7 different genotypes (M1-M7), in which humans have 3 type (M1-M3), M4 and M7 found only in birds, VP3 genes of TB-Chen strains belong to type M2; different VP3 genotypes have unique sensitive host range, the same genotype infection may not Different genotypes can also be infected in the same animal and the same animal. The results provide an important basis for the preparation of RV VP3 protein and further study of its structure and function.

【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R373

【参考文献】