作者：周李华, 叶德萍, 王智, 唐祥凯

作者单位：中国测试技术研究院, 四川 成都 610021

关键词：T4 DNA连接酶; 酶活力; 测定

摘要：

工具酶的酶活力检测是工具酶产品质量控制的关键,研究和建立工具酶酶活力的测定方法是工具酶产品标准化制定的重点和难点。该文采用改进的粘性末端单位测定T4 DNA连接酶酶活力。经验证,方法重复性好,稳定性佳,为建立T4 DNA连接酶质量检测标准奠定了基础。

Detection methods of activity of T4 DNA ligase

ZHOU Li-hua, YE De-ping, WANG Zhi, TANG Xiang-kai

National Institute of Measurement and Testing Technology, Chengdu 610021, China

Abstract: The key to the quality control of enzyme products is enzyme activity detection. And studying and establishing the method for determining the enzyme activity is the key of enzyme products standardization formulation. The improved sticky end unit method is investigated in this paper, which lays a foundation for the standard test methods of T4 DNA ligase quality.

Keywords: T4 DNA ligase; activity; determination

2014, 40(2): 64-66,81  收稿日期: 2013-6-17;收到修改稿日期: 2013-8-7

基金项目: 国家科技支撑计划项目(2012BAK25B01);公益性行业科研专项基金(201310240)

作者简介: 周李华(1978-),女,湖南茶陵县人,助理研究员,硕士,主要从事生命科学测试技术研究。

参考文献

[1] J·萨姆布鲁克,D·W·拉塞尔. 分子克隆实验指南上册[M]. 3版. 黄培堂,译. 北京:科学出版社,136.
[2] Song W, Pascal J M, Ellenberger T, et al. The DNA binding domain of human DNA ligas I interacts with both nicked DNA and the DNA sliding clamps[J]. DNA Repair,2009,8(8):912-919.
[3] Jia L J, Soengas M S, Sun Y. ROC1/RBX1 E3 ubiquitin ligase silencing suppresses tumor cell growth via sequential induction of G(2)-M arrest, apoptosis and senescence[J]. Cancer Research,2009,69(12):4974-4982.
[4] Weiss B, Jacqumin-Sablon A, Live T R, et al. Enzymatic breakage and joining of deoxyribonucleic acid.VI.Further purification and properties of polynucleotide ligase from Escherichia coli infected with bacteriophage T4[J]. Biol Chem,1968(243):4543-4555.
[5] 刘凌凤,唐志文,王柯敏,等. 基于分子信标实时监测大肠杆菌DNA连接酶催化的DNA连接过程[J]. 高等学校化学学报,2003,24(10):1761-1764.
[6] 刘斌,羊小海,王柯敏,等. 基于线性荧光探针对T4 DNA连接酶的活性分析[J]. 高等学校化学学报,2010,31(3):463-467.
[7] Sris K V, Shum A S. A second NAD(+)-dependent DNA ligas(LigB) in eschrichia coli[J]. Nucleic Acids Res,2001(29):4930-4934.