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Merck
CN

D2886

Sigma-Aldrich

DNA连接酶 来源于 T4-感染的大肠杆菌

buffered aqueous glycerol solution

别名:

T4 DNA 连接酶, 多核苷酸连接酶, 聚脱氧核糖核苷酸合酶

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About This Item

CAS号:
EC 号:
MDL编号:
UNSPSC代码:
12352204
NACRES:
NA.53

等级

for molecular biology

质量水平

形式

buffered aqueous glycerol solution

比活

4,000 U/mL

分子量

68 kDa

UniProt登记号

储存温度

−20°C

基因信息

bacteriophage T4 ... 30(1258680)

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应用

适用于:
  • 平端和粘性DNA片段的连接
  • 克隆载体和限制性插入片段的连接
  • 双链DNA和RNA或DNA/RNA杂合体中的缺口补平
  • 通过桥连寡核苷酸接头来偶联RNA单链

生化/生理作用

T4 DNA连接酶可在双链体DNA的相邻多核苷酸的末端之间形成能量依赖性的磷酸二酯键。连接反应需要ATP作为辅助因子。平末端片段的连接需要更高的酶浓度并可通过在反应混合物中使用PEG来促进。该酶需要3′羟基和5′磷酸基用于连接。可通过用碱性磷酸酶进行去磷酸化来防止载体DNA的自连。T4连接酶在DNA和RNA缺口的修复中可发挥积极作用。

组分

T4 DNA连接酶以溶于20 mM Tris-HCl(pH 7.5),50 mM KCl,1 mM DTT和50%(v/v)甘油的溶液形式提供。

单位定义

一个Weiss单位的定义是在37℃ 下,在20分钟内催化1 nmol的P32从焦磷酸盐交换至ATP作为Norit可吸收物质所需的酶量。

其他说明

T4 DNA连接酶可通过在65 °C加热10分钟灭活。

WGK

WGK 3

闪点(°F)

Not applicable

闪点(°C)

Not applicable

个人防护装备

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)

法规信息

常规特殊物品

分析证书(COA)

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Engler, M.J. and Richardson, C.C. et al.
The Enzymes, 5, 3-3 (1982)
Hiroshi Ochiai et al.
Proceedings of the National Academy of Sciences of the United States of America, 109(27), 10915-10920 (2012-06-20)
To understand complex biological systems, such as the development of multicellular organisms, it is important to characterize the gene expression dynamics. However, there is currently no universal technique for targeted insertion of reporter genes and quantitative imaging in multicellular model
M J Moore et al.
Science (New York, N.Y.), 256(5059), 992-997 (1992-05-15)
A simple and efficient method for synthesizing long, site-specifically modified RNA molecules was developed whereby segments of RNA were joined with the use of bacteriophage T4 DNA ligase. A single hydrogen or O-methyl group was substituted for the 2'-hydroxyl group
Xin Cheng et al.
European journal of cell biology, 91(10), 782-788 (2012-08-04)
Translocation of mitochondrial DNA (mtDNA) fragments to the nucleus and insertion of those fragments into nuclear DNA has been observed in several organisms ranging from yeast to plants and mammals. Disruption of specific nuclear genes by de novo insertions of
Wenxin Gu et al.
Bioorganic & medicinal chemistry letters, 22(11), 3693-3698 (2012-05-09)
A series of 2,6-disubstituted aminoalkoxypyrimidine carboxamides (AAPCs) with potent inhibition of bacterial NAD(+)-dependent DNA ligase was discovered through the use of structure-guided design. Two subsites in the NAD(+)-binding pocket were explored to modulate enzyme inhibitory potency: a hydrophobic selectivity region

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