Martsishauskas R P, Baronaĭte Z A, Sudzhiuvene O F, Pesliakas I G
Prikl Biokhim Mikrobiol. 1985 Jul-Aug;21(4):482-91.
The sorption capacity of the dye cibacron blue F3GA, immobilized on CL-Sepharose 6B and other support matrices, in respect to DNA- and RNA-ligases T4 was being studies. Cibacron blue F3GA immobilized on CL-Sepharose 6B binds a three-fold amount of DNA-ligase in comparison to RNA-ligase. The enzyme chromatography on cibbacron blue F3GA-CL-Sepharose 6B revealed a stronger linkage between DNA-ligase T4 and the sorbent than between RNA-ligase T4 and the sorbent. Elution was performed with potassium chloride. DNA-ligase T4 was eluted with 0.25-0.5 M KCl and RNA-ligase T4 with 0.08-0.18 M KCl. Since deoxyexonuclease contaminants possess stronger bonds with the sorbent than ligases, elution of deoxyexonucleases occurs at higher concentrations of KCl. Chromatography of enzymes on cibacron blue F3GA-CL-Sepharose 6B allows one to obtain DNA- and RNA-ligases essentially free of DNase and RNase contaminants.
对固定在交联琼脂糖6B和其他支持基质上的染料汽巴克隆蓝F3GA对T4 DNA连接酶和RNA连接酶的吸附能力进行了研究。与RNA连接酶相比，固定在交联琼脂糖6B上的汽巴克隆蓝F3GA结合的DNA连接酶量是其三倍。在汽巴克隆蓝F3GA-交联琼脂糖6B上进行的酶色谱分析表明，T4 DNA连接酶与吸附剂之间的连接比T4 RNA连接酶与吸附剂之间的连接更强。用氯化钾进行洗脱。T4 DNA连接酶用0.25 - 0.5 M KCl洗脱，T4 RNA连接酶用0.08 - 0.18 M KCl洗脱。由于脱氧外切核酸酶污染物与吸附剂的结合比连接酶更强，脱氧外切核酸酶在更高浓度的KCl下洗脱。在汽巴克隆蓝F3GA-交联琼脂糖6B上进行酶色谱分析可使人们获得基本不含DNase和RNase污染物的DNA连接酶和RNA连接酶。
