Cloning of toxic genes with mini-mu derivative of bacteriophage mu.
We describe an in vivo cloning method using mini-Mu phage for genes, which cannot be cloned on multicopy vectors, mainly for their toxicity. We have successfully cloned succinate dehydrogenase (sdh) gene E. coli which was inactivated with defined insertion of fragment Kmr by this method. The most of obtained Kmr clones of mini-Mu transductants have contained the sequence of whole sdh gene. The intact gene of sdh can be reconstructed by site-directed mutagenesis.
Stuchlík
S
S
Department of Molecular Biology, Comenius University, Faculty of Natural Sciences, Bratislava, Slovak Republic.
Janitorová
V
V
Turna
J
J
D010583:Bacteriophage mu / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial; D005822:Genetic Vectors; D007613:Kanamycin Resistance / Q000235:genetics; D016254:Mutagenesis, Insertional; D010957:Plasmids / Q000235:genetics; D013385:Succinate Dehydrogenase / Q000235:genetics
D013385:Succinate Dehydrogenase