耐辐射奇球菌基因组DNA表达文库的构建与鉴定  

Construction and Identificantion of Genomic DNA Expression Library for Deinococcus radiodurans R1

作　　者：徐向红[1] 李斌元[1] 马云[1] 廖永强[1] 何淑雅[1] 

机构地区：[1]南华大学生物化学与分子生物学教研室,湖南衡阳421001

出　　处：《现代生物医学进展》2009年第5期818-820,共3页Progress in Modern Biomedicine

基　　金：国家自然科学基金(30770647);湖南省自然科学基金(06JJ2023)

摘　　要：目的:构建耐辐射奇球菌(Deinococcus radioduransR1)基因组DNA表达文库,为进一步研究耐辐射奇球菌高抗辐射的调控网络奠定基础。方法:提取耐辐射奇球菌基因组DNA,用Sau3AI酶将基因组DNA部分酶切成0.5-5kb大小的片段,用T4DNA连接酶将部分酶切片段与经BamHⅠ和碱性磷酸酶(CIAP)处理的pGADT7载体进行连接后电击转化大肠杆菌DH5α。结果:得到重组子数为2.2×104,扩增后的文库滴度为108cfu/mL。结论:构建了耐辐射奇球菌基因组pGADT7表达文库,为进一步筛选与高抗辐射相关基因产物的互作蛋白奠定了基础。Objective： To construct genomic DNA expression library of Deinococcus radiodurans R1 for the research of its interaction network in anti-radiation related proteins. Methods： The genomic DNA was extracted, and digested with Sau3AI. The DNA fragments, most of which were 0.5-5 kb, were obtained subsequently. Then, these DNA fragments were linked to the vector pGADT7 treated with BamH I and CIAP, and then transformed into E.coli DH5α by electroporation methods. Results： The results showed that the library contained 2.2 × 10^4 clones and its titer was 108 cfu/mL. Conclusion： The genomic DNA expression library of Deinococcus radiodurans R1 has been constructed, which lays a foundation for further screening for the proteins that might interact with the anti-radiation related proteins from Deinococcus radiodurans R1.

关 键 词：耐辐射奇球菌 基因组DNA表达文库 酵母双杂交系统 

分 类 号：R378.1[医药卫生—病原生物学] Q75[医药卫生—基础医学]