摘要
通过鉴别寄主反应、病毒部分序列测定确定了采自广州白云区表现花叶、斑驳症状的节瓜上的病毒为ZYMV。采用RT PCR方法扩增和克隆了该病毒的外壳蛋白基因 ,连接到原核表达载体pET 2 2b(+)上。获得的重组子pET ZCP转化大肠杆菌BL2 1(DE3)后 ,用IPTG进行诱导表达。SDS PAGE和Westernblot分析表明 ,CP基因在大肠杆菌中获得了高效表达 ,融合蛋白分子量约为 33 0kD。将融合蛋白纯化后免疫兔子 ,获得了特异性较高的抗血清。ELISA测定其效价为 1 4 0
A Guangzhou isolate of ZYMV infecting Benincasa hispida Cogn. var. chieh-qua How was identified by indicator tests and partial sequence amplification. The coat protein (CP) gene of this virus was amplified by RT-PCR, and ligated to the expression vector pET-22b(+). The recombinant plasmid pET-ZCP was transformed into E.coli BL21(DE3) and then induced to express 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 33.0kD. Antiserum with high specificity was produced after the rabbit was immunized with purified recombinant protein, and the titer was determined to be 1/4096 by antigen coating plate-ELISA (ACP-ELISA).
出处
《微生物学报》
CAS
CSCD
北大核心
2005年第1期132-134,共3页
Acta Microbiologica Sinica
基金
科技基础性工作专项资金项目 ( 2 0 0 1DEA10 0 0 4)
农业部 948项目 ( 2 0 0 1-2 49)~~
