香蕉线条病毒ORF I基因的原核表达及抗体制备  

Prokaryotic Expression of Banana streak virus ORF I Gene and Preparation of Its Antiserum

作　　者：何云蔚[1] 陈秀[1] 阮小蕾[1] 沈文锦[1] 刘福秀[1] 李华平[1] 

机构地区：[1]华南农业大学植物病毒研究室,广东广州510642

出　　处：《华南农业大学学报》2009年第3期18-21,共4页Journal of South China Agricultural University

基　　金：国家自然科学基金(30671358);国家农业公益性行业科研专项(nyhyzx07-029)

摘　　要：利用PCR方法从含香蕉线条病毒广东分离物(BSV-GD)部分基因组的质粒中扩增该病毒的ORF I基因,将其克隆到pET-28b(+)原核表达载体中进行了融合表达.SDS-PAGE电泳发现,目的融合蛋白与预期大小一致,相对分子质量约为23 000,表达产物主要以不可溶的包涵体形式存在,将其变性溶解后利用N端的组氨酸标签进行纯化.以纯化产物为抗原免疫兔子制备了香蕉线条病毒的阳性兔抗血清,Western blot和ELISA分析表明,制备的兔抗血清为香蕉线条病毒的高效特异性抗血清,效价高达1∶51 200.ORF I gene of Banana streak virus Guangdong （BSV-GD） isolate was amplified from a BSVGD recombinant plasmid by PCR, and the gene was expressed by being cloned into prokaryote expression vector pET-28b（ ＋ ）. The interesting fusion protein was about 23 000 in size by SDS-PAGE analysis. The product was existed in form of inclusion body. It was dissolved by denaturalization, and the purified protein was obtained by using the histidine labeling kit of N-terminus of the protein. The antiserum was obtained .by immunizing healthy rabbits with the purified protein. The results from Western blot and ELISA analysis showed that it＇ s a special antiserum of BSV with high titer which was determined to be 1：51 200. This study was a base of the research of BSV including ORF I gene＇ s function and the virus detection.

关 键 词：香蕉线条病毒 ORF I 原核表达 抗血清 

分 类 号：S432.1[农业科学—植物病理学]