摘要
利用PCR技术将鸡痘病毒env基因亚克隆到pVAX1真核载体上,构建了含有该基因的真核表达质粒pVAX-env。将质粒pVAX-env体外瞬时转染BHK21细胞通过间接免疫荧光检测蛋白表达。将pVAX-env质粒肌肉注射免疫14日龄雏鸡,通过RT-PCR和组织免疫荧光检测体内质粒表达。结果表明,成功构建了真核表达质粒pVAX-env,并证明pVAX-env在体内体外都能有效表达。研究为鸡痘病毒基因工程疫苗进一步研制及应用奠定了基础。
The env gene of Fowpox virus（FPV） Chinese isolate was cloned and inserted into the expression vector pVAX1,resulting in a recombinant expression plasmid pVAX-env.Subsequently,the plasmid pVAX-env was transfected into BHK21 cells.The expression of env protein was detected by immunofluorescence assays.Then,the same plasmid was injected into chickens,and the eukaryotic expression of the protein was further confirmed by RT-PCR and immunohistochemical fluorescence.The results showed that eukaryotic expression plasmid pVAX-env was successfully constructed,and pVAX-env was able to be expressed in vitro and in vivo.This research paved the way for the further study and application of FPV genetic engineering vaccine.
出处
《东北农业大学学报》
CAS
CSCD
北大核心
2011年第6期105-108,I0002,共5页
Journal of Northeast Agricultural University
基金
黑龙江省自然科学基金重点项目(ZJN0702-01)
关键词
鸡痘病毒
ENV
真核表达
Fowlpox virus
env
eukaryotic expression
