Kanaya E, Uchiyama Y, Ohtsuka E, Ueno Y, Ikehara M, Kanaya S
Protein Engineering Research Institute, Osaka, Japan.
FEBS Lett. 1994 Nov 7;354(2):227-31. doi: 10.1016/0014-5793(94)01131-1.
A series of DNA-linked ribonucleases H with DNA adducts varying in size and sequence, ranging from heptamer to nonamer, were constructed and examined for their ability to cleave the 12-base RNA (5'-CGGAGAUGACGG-3') site-specifically. The DNA-linked RNase H with the 9-base DNA (5'-GTCATCTCC-3') cleaved the 12-base RNA specifically at A6-U7. Kinetic studies revealed that the DNA-linked RNase H with the 8-base DNA (5'-TCATCTCC-3') cleaved it slightly more effectively than that with the 9-base DNA. Factors that may affect the specificity and catalytic efficiency of a DNA-linked RNase H are described.
构建了一系列带有大小和序列各异的DNA加合物（从七聚体到九聚体）的DNA连接核糖核酸酶H，并检测它们对12个碱基的RNA（5'-CGGAGAUGACGG-3'）进行位点特异性切割的能力。带有9个碱基DNA（5'-GTCATCTCC-3'）的DNA连接核糖核酸酶H在A6-U7处特异性切割12个碱基的RNA。动力学研究表明，带有8个碱基DNA（5'-TCATCTCC-3'）的DNA连接核糖核酸酶H比带有9个碱基DNA的切割效率略高。描述了可能影响DNA连接核糖核酸酶H特异性和催化效率的因素。
