Rybakov V N, Bogachev V S
Bioorg Khim. 1989 Jun;15(6):796-9.
Joining of oligodeoxynucleotides by T4 polynucleotide ligase on an oligodeoxynucleotide template analogue having P--S--C(5') internucleotide bonds was investigated. The enzyme did ligate the oligodeoxynucleotides but with an extremely low rate. The major product of the enzymatic reaction proved to be the adenylylated intermediate of the 5'-phosphorylated oligodeoxynucleotide, the product of the second stage of the overall ligation reaction. It is noteworthy that this intermediate is not accumulated in ligase reactions under standard conditions when DNA templates rather than template analogues were used.
研究了T4多核苷酸连接酶在具有P-S-C(5')核苷酸间键的寡脱氧核苷酸模板类似物上连接寡脱氧核苷酸的情况。该酶确实能连接寡脱氧核苷酸,但速率极低。酶促反应的主要产物被证明是5'-磷酸化寡脱氧核苷酸的腺苷酸化中间体,这是整个连接反应第二阶段的产物。值得注意的是,当使用DNA模板而非模板类似物时,在标准条件下的连接酶反应中不会积累这种中间体。