Using Suppression Subtractive Hybridization as a Tool for Identifying Lipopolysaccharide (LPS) Induced Immunity Genes of Orange-Spotted Grouper (Epinephelus coioides) Macrophage

• Main Authors: Shih-JieLin, 林士傑
• Other Authors: Han-You Lin
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/37968593464170072614

碩士 === 國立成功大學 === 生物科技研究所碩博士班 === 98 === Epinephelus coioides (E. coioides, orange-spotted grouper) is an important cultivated fish in Taiwan, but various fish diseases cause many difficulties in aquaculture. How to control these diseases were the most crucial issues in the research of aquatic industry development. In fish biology, the immune system is highly related with the disease resistant ability. Due to the long development distance between fish and vertebrate animals, the fish immune system is quite different from human and mammal. One of the biggest differences is the nucleotide sequences of genes involved in immunology response and the variation of the genes was not only between fish and vertebrate, but also between different fish species. The researches in molecular mechanisms of fish immunity and the immunity genes are only in the initial stage. Suppression subtractive hybridization technology (SSH) is a high throughput method to screen for multiple immune related genes expressed in orange-spotted grouper macrophages under immune-stimulator, lipopolysaccharide (LPS) induction. According to NCBI blast analysis, 81 genes were identified in over 400 investigated SSH clones. There were 12 genes directly involved in immune response. We further analyzed the immunity gene expression under LPS stimulation. These preliminary data showed further insights to understanding the relation of cytokine performance and the immune system in the grouper, suggesting a proper and efficient strategy to vaccine development and disease prevention.