摘要
本研究将伪狂犬病病毒Bartha株基因组与含有LacZ标志基因的TK基因转移质粒 pUEKPZ共转染猪肾传代细胞PK 15 ,细胞出现病变后 ,反复冻融 3次收毒 ,按 1∶5稀释接种于IBRS 2细胞。在X gal存在下挑取蓝斑 ,蓝斑筛选 3次 ,再进行空斑试验 ,同时用PCR扩增LacZ基因 ,经 3次空斑纯化 ,随机挑取的空斑均能扩增出LacZ基因 ,证实所获得的重组病毒为伪狂犬病病毒Bartha株TK-/LacZ+ 突变株。TCID50 试验表明 ,TK失活对Bartha株在细胞上增殖无影响 ;Balb/C小鼠试验表明 。
In this study,the TK gene transfer vector pUEKPZ containing LacZ gene and PRV Bartha genome DNA were cotransfected onto PK 15 cells.When cytopathogenic effect appeared,virus contained supernatant was harvested.Transfection progeny was frozen and thawed for three times,diluted as 5 fold volume,and plated onto IBRS 2 cells at the presence of X gal.The picked blue plaques were purified by plaque technique for three times.Then plaques were picked randomly,used as templates and LacZ gene fragment could be amplified from all of them.This result showed that the mutant virus was PRV Bartha TK -/LacZ + strain.TCID 50 assay indicates the mutant strain grows well on PK 15 cells;Inoculation to Balb/C mice indicates it is safer to Balb/C mice than the parent virus Bartha.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2002年第3期304-307,共4页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
高等学校博士学科点专项科研基金资助
