美洲拟鲽抗冻蛋白基因植物表达载体的构建-基础兽医学专业毕业论文.docxVIP

李树峰 李树峰 摘要 2002年5月 摘 要 在生物体内已经发现了多种类型的抗冻蛋白，例如：鱼类抗冻蛋白、植物抗冻蛋白 和昆虫抗冻蛋白．其中研究比较清楚的是鱼类的抗冻蛋白，对植物和昆虫抗冻蛋白的研 究也正在开展．抗冻蛋白有很多重要的应用，它可以改进农作物的产量和质量，增强 幼苗的存活力。延长作物的生长季节和栽种范围，还可以降低作物的贮存温度， 增加贮存的时间。此外，抗冻蛋白在医学、食品科学、工业上也都有重要的应用。 本实验中人工合成了美洲拟鲽抗冻蛋白基因，并在美洲拟鲽抗冻蛋白基因两侧添 加了许多调控序列，如具有加倍增强子的35S启动子和nO$终止子，它们可以促 进抗冻蛋白基因的转录。及可在翻译水平上提高基因表达的。序列和Kozak序列， 从而构建了抗冻蛋白基因的高效表达盒．将此表达盒连接到pBll21．1上，最终获 得了抗冻蛋白基因植物高效表达载体pBRTSAFP。用基因枪法将之导入玉米的愈 伤组织中，井检测了与抗冻蛋白基因连在一起的gus基因的表达情况。检测结果 表明：gus基因瞬间表达十分强烈。 关键词：载体构建 抗冻蛋白 调控序列 2 李树峰 李树峰 Abstract 2002年5月 Construction of Plant Expression Vector Harboring Pseudopleuronectes americanus antifreeze protein gene Abstract The antifreeze proteins have been found in many organisms，such as fishes， plants and insects and the antifreeze proteins in fishes are studied in detail．The antifreeze proteins have many applications in increasing the quantity and quality of crop．improving the seedling liviability，prolonging the growth-season of crop and extending the planting·range，as well as decreasing the stored temperature of crop to prolong the stored time．Moreover，there are many important applications in medicine， foodstuff science and industry． In our experiment，a lot of regulator sequences including the 35S promoter with double enhancer elements and the terminator nos．the n sequence and Kozak sequence for improving the genetic statement in the level of translation，were appended on the two sides of the artifically Pseudopleuroneetes americanus antifreeze protein gene by a series of process．and the products were]inked into pBI】21．】to get the expression vector for the antifreeze protein gene followed by importing it into the caluus of maize by gene—gun．At last，we detected the expression of gus gene linked to antifreeze protein gene．The result indicates that gus gene is expressed strongly and instantaneousIy in caluus of maize． Key word：carrier design antifreeze protein regulator sequence 耋丝竺一。=：：。：：；。：：==：=：=：。墼：：：：：：!。：：一一一型兰些1前 耋丝竺一。=：：。：：；。：：==：=：=：。墼：：：：：：!。：：一一一型兰些 1前 言 抗冻蛋白(ami6ceze