摘要
本实验用RT$CPCR的方法获得李属坏死环斑病毒的cp基因,并将其连接到表达载体pET-22b(+)上,得到重组质粒pETPNRSVCP,转化大肠杆菌BL21(DE3)后,用IPTG进行诱导表达。SDSPAGE和Westernblot分析结果表明,cp基因在大肠杆菌中获得了高效表达,获得的融合蛋白分子量约为29.0kDa。将该融合蛋白免疫兔子,获得PNRSV特异性抗血清。酶联法(ELISA)测定的效价为1/1024。为用血清学方法检测该病毒提供了基础条件。
The coat protein （CP） gene of Prunus necrotic ringspot virus （PNRSV） was amplified by RT-PCR, and ligated to the expression vector pET-22b （ ＋ ）. The recombinant plasmid pET-WVMVCP was transformed into E. coli BL2.1 （DE3） and then induced by IPTG. It was showed that the CP gene was highly expressed by SDS-PAGE and Western blot analysis. The molecular weight of the recombinant protein was about 29.0 kDa. Antiserum with high specificity was produced after the rabbit was immunized with purified reeoinbinant protein, and the titer was determined to be 1/1024 by antigen coating plate- EHSA（ACP-ELISA）. It provides the basic condition for serological detection of PNRSV.
出处
《植物检疫》
2006年第2期73-75,共3页
Plant Quarantine
基金
农业部948资助课题(植物检疫性有害生物检疫试材及技术2001-249)
