摘要
从猪瘟病毒石门株血液样品中提取总RNA,通过RT-PCR对Erns基因进行cDNA扩增,获得了696bp的片段。将该片段克隆于T-easy载体后进行序列分析,确认PCR产物为猪瘟病毒Erns基因,从阳性克隆中提取质粒,经BamHⅠ和HindⅢ双酶切,回收产物亚克隆到pET-32a表达载体中,提取质粒后转化BL21(DE3)感受态细胞,并筛选出阳性克隆,经IPTG诱导后通过SDS-PAGE检测出Erns基因的表达。
E^rns gene fragments of Classical swine fever virus Shimen strain were amplified by RT-PCR from total RNA of blood sample, purified products were cloned into T-easy vector. After identification with enzyme digestion and sequence analysis, E^rns gene was subcloned into transfer vector PET32a,the recombinant plasmid was obtained after selection and then transformed into the host E. coli strain BL21 （DE3）for expression. Induced by lmmol/L IPTG at 27℃ ,the expression product of this gene was identified by SDSPAGE. The results revealed that E^rms protein had been expressed successfully.
出处
《动物医学进展》
CSCD
2007年第2期22-24,共3页
Progress In Veterinary Medicine
关键词
猪瘟病毒
E^rns蛋白
原核表达
Classical swine fever virus
E^rns protein
prokaryotic expression
