The molecular cloning and characterization of cobia（Rachycentron canadum）immunoglobulin gene

• Main Authors: Wen-Pin Chung, 鍾文彬
• Other Authors: Lin Cheng-Hui
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/24187333590868047904

碩士 === 國立臺灣海洋大學 === 水產養殖學系 === 93 === The immunoglobulin (Ig) is distributed and located in blood, mucus, etc. when immunoglobulin combine to antigen can induce subsequence immune response. Therefore, it play an important role in homoral immunity. The immunoglobulin is mainly of IgM in the teleost. In this study, we have isolated two full length immunoglobulin heavy chain of cobia (RC8、RC10) from headkidney which containing 1926 and 1923 bp, both deduced 583 amino acid, and the molecular weight are about 65 kDa. Four full length of immunoglobulin light chain (RC12、RC17、RC21、RC31) is containing 926, 966, 1009 and 1006 bp, respectively. It’s can deduced 236, 244, 241 and 241 amino acid, and the molecular weight are about 23 kDa. The molecular weight of immunoglobulin tetramer nearly as 696 kDa. In protein purification, we utilize ammonium sulfate precipitation and gel extraction to isolate cobia IgM. Utilizating SDS-PAGE to estimate the molecular weight heavy chain is about 69 kDa and light chain is about 30 kDa, the molecular weight of immunoglobulin tetramer nearly as 792 kDa. The produce antiserum of antibody via immune rabbit. Analysis of Western blotting which antibody of rabbit, it can detected the immunoglobulin of cobia.