精品文档---下载后可任意编辑鱼源Ⅲ型抗冻蛋白基因的原核表达及转革胡子鲶鱼方法模型的建立的开题报告摘要抗冻蛋白是一种能够使生物在低温环境下存活的蛋白质，是一种重要的应对低温逆境的生物分子。本讨论旨在通过原核表达及转革胡子鲶鱼方法模型的建立讨论鱼源Ⅲ型抗冻蛋白基因的功能。讨论将在胡子鲶鱼（Silurus asotus）中进行。首先通过 PCR 法从胡子鲶鱼的基因组中扩增鱼源Ⅲ型抗冻蛋白基因，并对其进行序列分析和比对。然后将鱼源Ⅲ型抗冻蛋白基因进行亚克隆，将其插入到原核表达载体 pET-28a 中，然后转化到大肠杆菌 BL21 (DE3)中，进行原核表达。利用 Western blot 和 SDS-PAGE 技术验证表达产物并纯化蛋白。最后，将该基因构建植入载体并植入到胡子鲶鱼中，讨论其在胡子鲶鱼中的功能及作用机制。关键词：抗冻蛋白，鱼源Ⅲ型，原核表达，转革胡子鲶鱼AbstractAntifreeze protein is a type of protein that helps living organisms survive in low temperature environments, and it is an important biological molecule to cope with low temperature stress. The aim of this study is to research the function of fish antifreeze protein III gene through the establishment of a prokaryotic expression and transgenic Silurus asotus model. The study will be conducted in Silurus asotus. First, the fish antifreeze protein III gene is amplified by PCR from the genomic DNA of Silurus asotus, and then its sequence is analyzed and aligned. The fish antifreeze protein III gene is then subcloned and inserted into the prokaryotic expression vector pET-28a, and transformed into Escherichia coli BL21(DE3) for prokaryotic expression. The expression product is verified by Western blot and SDS-PAGE, and the protein is purified. Finally, the gene is constructed and implanted into Silurus asotus to study its function and mechanism in Silurus asotus.Keywords: antifreeze protein, fish type III, prokaryotic expression, transgenic Silurus asotus
