文献类型：期刊文献

中文题名：BLyS改组噬菌体文库的构建及初步筛选

英文题名：Change of Binding Activity between BLyS and Its Receptor BCMA by DNA Shuffling

作者：沈琼[1];宋聿文[1];韩威[2];丁艳丽[2];龚毅[2];袁勤生[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237;[2]中国科学院上海中科伍佰豪生物工程有限公司,上海200233

年份：2006

卷号：26

期号：5

起止页码：49

中文期刊名：微生物学杂志

外文期刊名：Journal of Microbiology

收录：CSTPCD;;CSCD:【CSCD_E2011_2012】；

语种：中文

中文关键词：B细胞刺激引子;DNA改组;噬菌体展示;文库

外文关键词：BLyS; DNA shuffling; phage display; library

摘要：从质粒pT7474-BLyS及人胎脑cDNA文库中分别扩增出BLyS和APRIL基因,用DNase I消化后,回收小于50 bp的片断,用于DNA改组。PCR产物与噬菌体载体pfUSE5连接后,电转E.coliER2738获得改组文库。构建的改组文库库容量为8.9×105。对文库进行初步的筛选,获得了一个受体结合活性降低的突变克隆。成功构建了BLyS改组噬菌体文库,为蛋白结构与功能之间关系的深入研究打下了基础。
BLyS （B lymphocyte stimulator） and APRIL genes were amplified from plasmid pT7474-BLyS and human fetal brain cDNA library respectively. After digested with DNase I, the fragments less than 50 bp were purified and reassembled to mutated BLyS genes by DNA shuffling. Their PCR products were inserted into vector pfUSE5 to establish a shuffled library of BLyS genes. The shuffled library consisted of about 8.9×10^5 clones. The library was primarily screened and one clone with lower activity was obtained. Therefore, the shuffled library of BLyS has been successfully constructed. This strategy also provided useful information on the study of structure-function relationship of proteins.