Vaisvila R, Sliesaraviciute Z, Kulakauskas S, Janulaitis A
Institute of Biotechnology FERMENTAS, Vilnius, Lithuania.
Gene. 1995 May 19;157(1-2):55-7. doi: 10.1016/0378-1119(94)00792-q.
A genetic system enabling the in vivo selection of genes encoding the DNA-modifying enzymes was developed. A gene library is transformed into a strain harboring the restriction-modification (R-M) system which a recognition sequence is a subset of the target sequence of the DNA methyltransferase (MTase) to be cloned. If the residing MTase is temperature sensitive, the inability of transformants to grow at 42 degrees C provides a simple and convenient procedure for the isolation of new MTase-encoding genes. The feasibility of this procedure has been demonstrated by the isolation of the ppu21IM gene from a Pseudomonas putida RFL21 gene library.
开发了一种能够在体内选择编码DNA修饰酶基因的遗传系统。将一个基因文库转化到一个含有限制修饰（R-M）系统的菌株中，该系统的识别序列是待克隆的DNA甲基转移酶（MTase）靶序列的一个子集。如果存在的MTase对温度敏感，转化子在42℃下无法生长，这为分离新的编码MTase的基因提供了一种简单便捷的方法。通过从恶臭假单胞菌RFL21基因文库中分离出ppu21IM基因，证明了该方法的可行性。
