热带作物学报2009，Vol.30Issue(8)：1113-1119,7.
建兰花叶病毒CP基因的原核表达及抗血清制备
Prokaryotic Expression and Antiserum Preparation of the Coat Protein of Cymbidium Mosaic Virus
摘要
Abstract
Cymbidium mosaic virus (CyMV) is one of the most important and worldwide viruses attacking orchids. This virus causes the symptoms of mosaic,chlorosis,necrosis and malformation in the orchids,and has a high economic impact to the orchid industry. Cattleya plants contracted with a disease were collected as samples from Zhangzhou,Fujian,and were identified to be infected with Cymbidium mosaic virus by using ID -ELISA and electronic microscopy assay. One pair of specific primers was designed for amplification of the coat protein(CP) gene from the samples infected with CyMV. The open reading frame encoding CP of CyMV isolate obtained from Zhangzhou,Fujian is 672 bp,encoding a 23.6 ku protein with 223 aa. The expected CP gene was then inserted into the pET-29a(+)vector for prokaryotic expression. And the aimed protein was purified and used to immune the rabbit for antiserum preparation. According the result of ID-ELISA analysis,specific rabbit anti-CyMV serum was prepared with a high titre of 1:51 200,a working concentration of 1:1 000 and sap sensitivity of 0.39 mg/mL. Western blot analysis confirmed that the antiserum reacted strongly and specifically to the CP of CyMV. There were no cross reactions between the antiserums and 11 species of homologous or heterologous viruses such as tobacco mosaic virus.关键词
建兰花叶病毒/外壳蛋白/原核表达/抗血清Key words
Cymbidium mosaic virus/coat protein/prokaryotic expression/antiserum分类
农业科技引用本文复制引用
罗金水..建兰花叶病毒CP基因的原核表达及抗血清制备[J].热带作物学报,2009,30(8):1113-1119,7.基金项目
福建省重大专项前期研究项目(No.2005N1004)资助. （No.2005N1004）
