摘要
从GenBank上下载鸡传染性支气管炎病毒(IBV)H120型标准株M基因的标准序列,以此标准序列为模板设计合成一对引物。从IBV尿囊液中提取总RNA,采用PCR技术扩增出M基因,将扩增出的目的片段克隆至pMD18-T载体上获得pMD18-IBV-M质粒,通过亚克隆将目的基因连接到经过改造的pEGFP-C1-SCP-SEC载体上,成功构建真核表达质粒pC1-IBV-M。通过间接免疫荧光试验验证了构建的真核载体能在真核细胞内表达,通过动物免疫试验来检测该真核表达载体对动物的免疫保护情况。
According to the M gene sequence of chicken infectious bronchitis H120, the primers were designed and synthesized. The total RNA was acquired from allantois fluid, M gene was amplified by using PCR technology. The plasmid of pMD18-IBV-M was cloned and then the M gent was inserted into pEGFP-C1-SCP- SEC vector which has been modified to construct the eukaryotic expression vector pCI-IBV-M. The animal＇ s immune protection was detected by indirect immunofluorscence（IFA） and animal experiment.
出处
《上海农业学报》
CSCD
2017年第2期104-108,共5页
Acta Agriculturae Shanghai
基金
上海市科技人才计划(14YF1414600)
上海市农业科学院中青年科技人员"助跑"计划
