不同代次缺失meq基因的重组马立克病毒SC9-1株主要基因序列同源性分析

发布时间：2018-05-03 05:30

本文选题：马立克病毒 + SC-　；参考：《中国兽医学报》2017年05期

【摘要】：分析缺失meq基因的重组马立克病病毒SC9-1原代、10代及40代主要基因序列的同源性。提取缺失meq基因的重组马立克病病毒SC9-1原代、10代及40代DNA,设计引物PCR扩增其与致瘤相关pp38基因、缺失meq基因后残余的FRT位点序列及囊膜蛋白基因gB、gC、gD、gE、gH、gI、gK,连接pMD18-T载体,转化DH5α感受态细胞,挑取阳性克隆进行测序,DNAStar软件对3代次各基因片段进行同源性比对。不同代次SC9-1病毒株的致瘤相关pp38基因、残留FRT位点序列及囊膜蛋白基因gB、gC、gD、gE、gH、gI、gK的核苷酸序列同源性均为100%,核苷酸序列差异性分析结果显示序列完全一致,无位点上的变化。缺失meq基因的重组马立克病病毒SC9-1在鸡胚成纤维细胞(CEF)上的传代过程中其致瘤相关pp38基因、残留FRT位点序列及囊膜蛋白基因gB、gC、gD、gE、gH、gI、gK没有发生变异,侧面反映了SC9-1在CEF细胞传代过程中具有很好的遗传稳定性。
[Abstract]:To analyze the homology of the main gene sequences of the recombinant Marek disease virus (SC9-1) with deletion of meq gene in 10 and 40 generations of Marek's disease virus (Marek's disease virus). The recombinant SC9-1 of Marek's disease virus (Marek's disease virus) with deletion of meq gene was extracted from the 10 and 40 generations of SC9-1. Primers were designed to amplify the pp38 gene associated with tumorigenesis. The remaining FRT loci of meq gene and the envelope protein gene (gBGCU gDgEgEgHngHngGK) were sequenced and ligated to the pMD18-T vector, and the DNA of Marek's disease virus (Marek's disease virus) was amplified by PCR. The positive clones were transformed into DH5 伪 competent cells and sequenced by DNA Star software to compare the homology of three generations of gene fragments. The nucleotide sequence homology of the oncogenic pp38 gene, the residual FRT locus, and the envelope protein gene (gBGCngDG) gEG HG Ig K of different generations of SC9-1 virus strains were 100 and 100, respectively. The results of nucleotide sequence difference analysis showed that the sequence was completely consistent, and there was no change in the nucleotide sequence. During the passage of recombinant Marek disease virus (SC9-1) with deletion of meq gene in chicken embryo fibroblast, its tumor-associated pp38 gene, residual FRT locus sequence and envelope protein gene gBGCngDGDG EG HG Ig K were not mutated. The results showed that SC9-1 had good genetic stability in the passage of CEF cells.
【作者单位】：山东农业大学动物科技学院;
【基金】：国家自然科学基金青年科学基金资助项目(31402235)
【分类号】：S852.65

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