A suitable method for construction and cloning hybrid plasmids containing EcoRI-fragments of E. coli genome.
A convenient procedure for the isolation of specific EcoRI-fragments of E. coli genome and their amplification on Km-resistance plasmid victor CK delta11 is described. The hybrid molecules were constructued in vitro using EcoRI-digestion, followed by ligation. Then appropriated E. coli strain was transformed with ligated DNA mixture and hybrid plasmids CK delta11-arg+, CK delta11-his+, CK delta11-thr+ and CK delta11-leu+ containing loci of E. coli genome were selected by molecular cloning. The hybrid plasmids obtained consisted of one EcoRI-fragment of initial plasmid CK delta11 and one respective specific portion of E. coli genome.
D001120:Arginine / Q000378:metabolism; D004261:DNA Replication; D004926:Escherichia coli; D005111:Extrachromosomal Inheritance; D005796:Genes; D005821:Genetic Techniques; D006639:Histidine / Q000378:metabolism; D006824:Hybridization, Genetic; D007930:Leucine / Q000378:metabolism; D010957:Plasmids; D013912:Threonine / Q000378:metabolism
D013912:Threonine; D006639:Histidine; D001120:Arginine; D007930:Leucine