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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。
In order to investigate whether the meq gene-deleted Marek’s disease vaccine strain SC9-1 and the super-virulent strain Md5 were able to obtain the meq gene by natural recombination, SC9-1 and Md5 viruses were co-infected with chicken embryo fibroblasts CEF) and passed three generations on CEF continuously to extract single plaque virus DNA. At the same time, Md5 virulent virulent SPF chickens immunized with SC9-1 vaccine strain were isolated at different time points to extract single plaque virus DNA. The virus DNA obtained by the two methods was verified by PCR. The FRT residues were cloned and sequenced, and their homologies were compared. The viruses identified by either method were either SC9-1 or Md5, no recombinant virus was detected, and the FRT residue sequence homology was 100%. Results SC9-1 did not obtain the missing meq gene from the wild strain Md5, and the meq gene knockout region had good genetic stability.