9个PstI接头片段的合成及其酶反应研究

9个PstI接头片段的合成及其酶反应研究 THE SYNTHESIS OF NINE KINDS OF OLIGODEOXYNUCLEOTIDE CONTAINING THE RECOGNITION SEQUENCE FOR PstI AND THEIR INTERACTION WITH ENZYME 程琳 Chen Lin 1 first-author 邹国林 Zou Guolin 1 曹新文 Cao Xinwen 1 武汉大学生命科学学院 武汉大学生命科学学院 College of Life Sciences,Wuhan University College of Life Sciences,Wuhan University 采用固相亚磷酸三酯法和化学与酶促相结合的方法合成了９个ＰｓｔＩ接头片段，并对它们进行了酶切反应研究．结果表明：ＰｓｔＩ识别序列中尿苷的存在只降低其互补链的切割程度，对其本链没有影响；ＰｓｔＩ切割底物是按两步单股切割机制进行，作用的最小底物是含有识别序列的８～１２聚脱氧核糖核苷酸，且对底物中识别序列两边的碱基对数目有一定要求；ＰｓｔＩ识别序列中的二个胞苷在酶反应中起的作用是不一样的 In this paper,nine kinds of oligodeoxynucleotide containing the recognition sequence of restriction endonuclease PstI have been synthesized by a solid phase phosphite triester method or a combination of chemical and enzymatic methods.The results of their enzymatic hydrolysis show that the existence of urine in the recognition sequence of PstI has little influence on the cleavage of this strand,but reduces the cleavage rate of its complementary strand;PstI cleaves its substrate through the single strand cleavage with two steps,and the minimum PstI substrate is an eight to twelve oligodeoxynucleotide containing ist recognition sequence,and there is a requirement to the base pair numbers on both sides of Pst Ⅰ recognition sequence;two cytidine in the recognition sequence of PstI dont play the same role to enzymatic hydrolysis. 限制性核酸内切酶 固相亚磷酸三酯法 接头片段 催化机制 restriction endonuclease solid phase phosphite triester method oligodeoxynucleotide containing PstI recognition sequence catalytic mechanism O621.25 国家自然科学基金(61672102) 1996-04-01 2021-04-01 4