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。
To analyze the homology of the major gene sequences of the meq gene-deleted recombi nant Marek＇s disease virus strain 8C9-1 in the original, the tenth and the forty generations, the DNA of the three different generations of the meq gene-deleted recombinant Marek＇s disease virus strain SC9-1 was extracted,and the tumor associated gene pp38 and residual FRT sequence after the meq gene deleted, and envelope protein gene gB,gg,gD,gE,gH,gI,gK were amplified by PCR,then inserted into pMD18-T vector and transformed into DH5u, positive clones were se- quenced and the three generations of each gene fragment were analyzed for homologous compari- son by DNAStar software. Nucleotide sequence homology of the tumor associated gene pp38, resid- ual FRT sequence and envelope protein gene gB, gC, gD, gE, gH, gI, gK in different generations of SC9-1 strains was 100% ,nucleotide sequence difference analysis showed that the sequence was completely consistent with each other and no changed in any sites. The tumor associated gene pp38,residual FRT sequence and envelope protein gene gB, gC, gD, gE, gH, gI, gK of the meq gene-deleted recombinant Marek＇s disease virus strain SC9-1 have no sign of genetic mutation in the process of CEF cell passaging. It reflects that the SC9-1 has good genetic stability during CEF cell passaging from a side.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2017年第5期804-810,共7页
Chinese Journal of Veterinary Science
基金
国家自然科学基金青年科学基金资助项目(31402235)
作者简介
陈俊霞（1991-），女，硕士。
通讯作者，E—mail：ssu6307@163．com
E-mail：zzcui@sdau．edu．cn
