In vivo cloning of DNA into multicopy cosmids by mini-Mu-cosduction.
A general in vivo procedure for cloning Escherichia coli genes into cosmids has been developed. The method we describe here uses a deleted Mu phage (a mini-Mu) to transpose E. coli genes into cosmids during mini-Mu replication. The resulting cosmids clones are packaged in vivo into lambda phage particles. Plasmids carrying a particular DNA sequence can be selectively recovered after infection of a new host with the in vivo constructed genomic cosmid library. This system was used successfully to clone several E. coli genes.
D010582:Bacteriophage lambda / Q000235:genetics; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D003090:Coliphages / Q000235:genetics*; D003360:Cosmids; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005838:Genotype; D014161:Transduction, Genetic