摘要
为构建猪伪狂犬病毒BarthaK61株TK缺失株,利用PCR法从猪伪狂犬病毒BarthaK61株基因组分别扩增出TK基因的左右2条臂TKR和TKL,并将TKR和TKL定向插入到pQCXIX载体中,构建pTK-转移质粒.进一步将EGFP基因插入到pTK-质粒中,构建pTK-/EGFP转移质粒.用pTK-/EGFP质粒与伪狂犬病毒BarthaK61株基因组共转染BHK-21细胞,通过挑取病变荧光蚀斑的方法获得了PRV/gE-/TK-/EGFP重组病毒.
To construct TK-null recombinant pseudorabies virus （PRV）BarthaK6 1 strain as a technol-ogy support of the research of genetic engineering vaccine of pseudorabies virus.Transfer vector pTK- was constructed using homology recombination arms of PRV TK gene,which were successfully amplified by PCR from PRV BarthaK6 1 genome.Subsequently enhanced green fluorescent protein gene （EGFP）was cloned into pTK-,resulting a universal transfer vector pTK-/EGFP.Co-transfecting pTK-/EGFP and genome of PRV BarthaK61 into BHK-21cells,recombinant PRV/gE-/TK-/EGFP were obtained follow-ing plaque screening.
出处
《甘肃农业大学学报》
CAS
CSCD
北大核心
2013年第6期13-18,共6页
Journal of Gansu Agricultural University
基金
甘肃省科技支撑计划(1011NKCA052)
甘肃省农业生物技术研究专项(GNSW-2012-16)
家畜疫病病原生物学国家重点实验室自主研究课题
