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Cloning of the ppu21IM gene using a in vivo selection method

R Vaisvila¹, Z Sliesaraviciute, S Kulakauskas

¹Institute of Biotechnology FERMENTAS, Vilnius, Lithuania.

|May 19, 1995

Abstract:

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.

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