T4DNA连接酶对电转化效率的影响  

Effects of T4 DNA ligase on the electroporation

作　　者：王福利[1] 赵爱志[2] 韩月恒[3,4] 温伟红[3,4] 刘建平[1] 董青川[1] 秦卫军[1] 

机构地区：[1]第四军医大学西京医院泌尿外科,陕西西安710032 [2]第四军医大学西京医院肝胆外科,陕西西安710032 [3]第四军医大学西京医院 [4]第四军医大学基础部免疫学教研室,陕西西安710032

出　　处：《中国现代医学杂志》2011年第17期1974-1976,共3页China Journal of Modern Medicine

基　　金：陕西省自然科学基础研究计划面上项目(No:2009JM4003-2)

摘　　要：目的探讨T4DNA连接酶影响电转化效率的机制并寻找提高电转化效率的方法。方法将1μg噬粒pDAN5加入到连接反应体系中并进行沉淀、热灭活以及酚氯仿抽提等各种处理后进行电转化。结果热灭活、酚氯仿抽提、热灭活后沉淀及热灭活后酚氯仿抽提处理后的转化效率最高(4.6×108￣5.3×108),其次是沉淀处理(4.8×107),未处理的转化效率最低(4.4×106)。结论 T4DNA连接酶的活性是导致电转化效率降低的主要原因,任何可以灭活该酶活性的方法均可显著地提高转化效率。[Objective] To investigate the mechanism that T4 DNA liagse lower the electroporation efficiency and look for the way to improve it. [Methods] 1μg pDAN5 was added in the ligation system. Then after several different treatments respectively, electroporations were performed. [Results] The electroporation effieiencies of the liagtion mixture treated with heat inactivation, phenol-chloroform or precipitation after heat activation were the highest （4.6×10^8-5.3×10^8）, the lower was treated by precipitation only （4.8×107） and the lowest was not treated at all. [Conclusion] It is the activation of the T4 DNA ligase that lowered the electroporation efficiency and any method that could inactive it could improve the efficiency.

关 键 词：T4 DNA连接酶 电转化 

分 类 号：Q523[生物学—生物化学]