摘要
利用PCR技术扩增出IBDV的结构蛋白VP3基因,将其克隆到表达载体pPROEX-HTa中,获得重组质粒命名为VP3/PA,经PCR、酶切和序列分析鉴定表明,插入的位置、大小和读码框均正确。SDS-PAGE检测表明,经重组质粒VP3/PA转化、诱导的受体菌E.coliDH5α能表达结构蛋白VP3基因,约33Ku,表达量达到了菌体总蛋白的33.9%,经Westernblot等检测表明,诱导表达的抗原蛋白能与vvIBDV阳性血清发生特异性反应。这为IBDV血清学诊断方法的建立打下了基础。
By PCR, a structural protein gene VP3 of infectious bursal disease virus （IBDV） was determined from segment A of IBDV .The gene was cloned into expressing vector pPROEX-HTa.The recombinant plasmid named VP3/PA was constructed.The VP3/PA was used to transform into E.coli DH5α.The results of SDS-PAGE and Western blot indicated that the VP3 protein was expressed in high level （33.9 % ） and the recombinant fusion protein,which was about 33 Ku,had immunological reactive activity.This study lay on foundation for the development of the diagnosis methods in serology for IBDV.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2006年第2期124-127,共4页
Chinese Journal of Preventive Veterinary Medicine
关键词
传染性法氏囊病病毒
结构蛋白VP3基因
克隆与原核表达
infectious bursal disease virus （IBDV）
structural protein VP3 gene
cloning and prokaryotic expression
