Shinozaki K, Okazaki T
Nucleic Acids Res. 1978 Nov;5(11):4245-61. doi: 10.1093/nar/5.11.4245.
T7 gene 6 exonuclease has been shown to have an RNase H activity as well as a double-strand specific DNase activity by the following experiments: The RNase H activity coelutes with the DNase activity from DEAE-cellulose, phosphocellulose, hydroxyapatite, and Sephadex G-200 columns. Gene 6 exonuclease specified by a T7 strain with a temperature sensitive mutation in gene 6 has an extremely heat-labile RNase H activity as well as a heat-labile DNase activity. T7 gene 6 exonuclease degrades the RNA region of a poly(A) . poly(dT) hybrid polymer exonucleolytically from the 5' terminus, releasing a ribonucleoside 5'-monophosphate product. When the RNA strand of a 0X174 RNA . DNA hybrid molecule synthesized with E. coli RNA polymerase is degraded, a ribonucleoside triphosphate is produced from the 5'-triphosphate terminus. Participation of T7 gene 6 exonuclease in the removal of primer RNA in discontinuous replication of T7 DNA is discussed.
通过以下实验表明,T7基因6核酸外切酶具有核糖核酸酶H活性以及双链特异性脱氧核糖核酸酶活性:核糖核酸酶H活性与脱氧核糖核酸酶活性在DEAE-纤维素、磷酸纤维素、羟基磷灰石和葡聚糖G-200柱上共同洗脱。由基因6发生温度敏感突变的T7菌株所指定的基因6核酸外切酶具有极不耐热的核糖核酸酶H活性以及不耐热的脱氧核糖核酸酶活性。T7基因6核酸外切酶从5'末端以外切方式降解聚(A)·聚(dT)杂交聚合物的RNA区域,释放出核糖核苷5'-单磷酸产物。当用大肠杆菌RNA聚合酶合成的0X174 RNA·DNA杂交分子的RNA链被降解时,从5'-三磷酸末端产生核糖核苷三磷酸。讨论了T7基因6核酸外切酶在T7 DNA不连续复制中参与引物RNA去除的情况。