Cloning of the ppu21IM gene using a in vivo selection method.
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.
Vaisvila
R
R
Institute of Biotechnology FERMENTAS, Vilnius, Lithuania.
Sliesaraviciute
Z
Z
Kulakauskas
S
S
Janulaitis
A
A
D001426:Bacterial Proteins; D003001:Cloning, Molecular / Q000379:methods; D015254:DNA Modification Methylases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D005798:Genes, Bacterial; D019811:Hydroxylamine; D006898:Hydroxylamines; D008780:Methyltransferases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D016296:Mutagenesis; D010957:Plasmids; D016958:Pseudomonas putida / Q000201:enzymology* / Q000235:genetics*; D015183:Restriction Mapping
D001426:Bacterial Proteins; D006898:Hydroxylamines; D019811:Hydroxylamine; D015254:DNA Modification Methylases; D008780:Methyltransferases; C094488:Ppu21IM methyltransferase, Pseudomonas