摘要
利用反转录_聚合酶链式反应 (ReverseTranscription_polymeraseChainReaction ,PT_PCR)技术扩增鸡传染性支气管炎病毒 (IBV)中国地方分离株的核蛋白基因 ,并对其序列进行了测定 ,结果表明该病毒株核蛋白基因长度为 12 3 0bp ,编码蛋白由4 0 9个氨基酸残基组成。与已发表的参考毒株进行核苷酸同源性比较 ,发现与澳大利亚群毒株的亲缘关系最为密切。在此基础上 ,本实验构建了杆状病毒重组转移载体pBlue_N ,将其与线性化的杆状病毒DNA共转染Sf9昆虫细胞 ,经过三轮蚀斑纯化和PCR鉴定 ,获得重组杆状病毒rBac_N。SDS_PAGE分析和Westernblot检测的结果表明核蛋白基因在重组杆状病毒感染的Sf9昆虫细胞内获得表达 ,融合蛋白表达量占细胞蛋白总量的 19.4
With the specific RT_PCR, the nucleocapsid (N) gene of a Infectious Bronchitis Virus(IBV) strain isolated from China was cloned and sequenced. Then a recombinant baculovirus containing the N gene was constructed in order to express the N protein. The expression of N protein in Sf9 cell was determined by Western blot with polyclonal antibodies against IBV. The infected cells were collected at different time post_infection and analyzed by SDS_PAGE. The result showed that the expressed product had a molecular weight of approximately 56kDa and the expression level of the recombinant protein was up to 19.4% of the total cell protein.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2001年第6期401-403,共3页
Chinese Journal of Preventive Veterinary Medicine
关键词
鸡传染性支气管炎病毒
核蛋白
基因序列
重组杆状病毒
表达
Avian infectious bronchitis virus
Nucleocapsid
Sequene
Recombinant Baculovirus System Expression
