【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。更多还原

【Abstract】 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 BamHⅠ and CIAP, and then transformed into E.coli DH5α by electroporation methods. Results: The results showed that the library contained 2.2×104 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.更多还原