Production of polyclonal antiserum against Cucurbit chlorotic yellows virus coat protein and application for diagnosis.

• Main Authors: Ya-Wen Chiang, 蔣雅文
• Other Authors: Mu-Kuei Chu
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/92179180044274881751

碩士 === 中華醫事科技大學 === 生物醫學研究所 === 100 === Taiwan is located in the subtropics, the climate quite favor to the melon growing, yields this crop becoming to the one of important economic crops in Taiwan. The annual melon crop planted area about 23,000 hectares, the main cultivation area consist of Tainan City, Chiayi County, Yunlin County. Usually the occurrence of disease is one of the limiting factors of crop growth, which are including viruses, bacteria and fungi diseases. In 2009 , one virus disease was discovered from Tainan City and the Yunlin County, the crop appear the massive etiolation on the tissues of the crop. After the investigation, this virus was identical to Cucurbit chlorotic yellows virus isolated from 2004, in Japan (CCYV). At that time, researchers didn’t pay attention to this virus disease. Now, this virus disease has been caused on the melon crop was affected by a serious economic impact. In this study, we want to cloning and expression the coat protein gene from CCYV virus to produce antiserum to identify CCYV on the fields. The results of CP gene sequencing of strains pGEMT-CP/DH5α that the length of the nucleic acid of the CP gene 735bp gene, gene translated into protein of 245 amino acids and a molecular weight of 28 kDa. For the mass production of coat protein from the CP gene recombinant clone pQE31-CP/M15, the protein expression analysis confirmed that the CP gene can be stable expression in the pQE system. In this study, the antiserum of CP can be utilized to identify the chlorotic yellowing virus disease by immunoassay and Immunohistochemistry stain.