马立克氏病毒meq基因缺失株SC9-1通过自然重组获得meq能力的分析

发布时间：2018-01-19 10:41

本文关键词： 马立克氏病病毒疫苗毒株SC- 超强毒株Md meq基因 PCR鉴定　出处：《病毒学报》2017年01期 　论文类型：期刊论文

【摘要】：为了探究meq基因缺失的马立克氏病毒疫苗株SC9-1与超强毒株Md5是否能够通过自然重组获得meq基因的能力,将SC9-1疫苗毒和Md5超强毒共同感染鸡胚成纤维细胞(CEF),并在CEF上连续传三代,提取单个蚀斑的病毒DNA。同时将Md5超强毒接种免疫过SC9-1疫苗株的SPF鸡,在不同的时间点分离病毒,提取单个蚀斑的病毒DNA。将两种方式获得的病毒DNA进行PCR验证,并将香啤酒重组酶位点(FRT)残留序列克隆测序,比较其同源性。两种方式鉴定的病毒均为SC9-1或是Md5,没有检测到重组病毒,而且FRT残留序列同源性为100%。结果 SC9-1没有从野生毒株Md5获得缺失的meq基因,而且meq基因敲除区具有很好的遗传稳定性。
[Abstract]:In order to investigate the ability of SC9-1 and Md5, the Marek's virus vaccine strain with missing meq gene, to obtain meq gene by natural recombination. The chicken embryo fibroblasts were co-infected with SC9-1 vaccine virus and Md5 supervirulent virus and passed on CEF for three consecutive generations. A single plaque virus DNA was extracted. At the same time, the SPF chicken vaccinated with SC9-1 vaccine was inoculated with Md5 supervirulent virus, and the virus was isolated at different time points. The single plaque virus DNA was extracted. The virus DNA obtained from two ways was verified by PCR, and the residual sequence of the recombinant enzyme site was cloned and sequenced. Comparing their homology, the two methods were identified as SC9-1 or MD5, and no recombinant virus was detected. The homology of FRT residue sequence was 1000.Results SC9-1 did not obtain the missing meq gene from wild strain Md5, and the meq gene knockout region had good genetic stability.
【作者单位】：山东农业大学动物医学院;
【基金】：国家自然科学基金青年基金项目(项目号:31402235);题目:Ⅰ型马立克氏病毒独特基因sorf2生物学功能的研究
【分类号】：S852.65
【正文快照】： 马立克氏病(Marek’s disease,MD)是由马立克氏病毒(Marek’s disease virus,MDV)引起的鸡的一种肿瘤性疾病[1]。MDV属于α疱疹病毒,主要分为致病性的Ⅰ型和无致病性的Ⅱ、Ⅲ型MDV[2]。MD是目前唯一可用疫苗预防的禽病毒性肿瘤疾病,近些年来随着MDV毒力的不断增强,相继出现了

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