南极中山站沉积物宏基因组文库的构建及碱性蛋白酶基因ACPRO001的克隆、表达和性质分析

• Main Author: 张映京
• Other Authors: 李庆阁, 曾润颖
• Format: Thesis
• Language: Chinese
• Published: 2009
• Subjects: 宏基因组; 碱性蛋白酶; 定向进化; metagenome; alkaline protease; directed evolution; Zhongshan; Zhongshan Station; Antarc*; Antarctica
• Online Access: http://dspace.xmu.edu.cn/handle/2288/46005

本文从南极中山站排污入海口（ZSS）沉积物样品中直接提取环境样品微生物总DNA，大小在48Kb以上。经过吹打调整DNA片段大小（到30-45Kb）后构建了ZScosmid宏基因组文库，获得了约6500个阳性克隆子。推算该DNA文库的总容量为228Mb，可以覆盖约10株类似E.coli菌株的总基因组。 采用蛋白酶活性筛选功能板筛选该文库我们得到了一个编号为1146的阳性克隆能够分解底物脱脂牛奶，在筛选平板上产生清晰的透明水解圈。DNA测序后分析表明该克隆外源DNA包含一个由425个氨基酸编码组成的蛋白质的完整的开放阅读框(ORF)，GC含量为50.6%。BLAST软件分析该外源DNA包含一个完. In this study, the metagenomic DNA were exrtracted and purified from a sediment sample core from ZSS. The samples ZSS was collected from the coast near the China Zhongshan Station.and the site ZSS was the sediment within the treated sewage effluent to the sea. The size of the DNA was more than 48Kb. Than the size was adjusted to 30-45Kb and was used to constract the metagenomic cosmid library. The. 学位：理学硕士 院系专业：生命科学学院生物医学科学系_细胞生物学 学号：21720061152199