CONSTRUCTION AND ANALYSIS OF RECOMBINANT LAMBDA-PHAGES CONTAINING MITOCHONDRIAL-DNA FRAGMENTS

Rat mtDNA has a molecular length of about 16 kilobase (kb) pairs and is cleaved into seven fragments by restriction endonuclease EcoRI. These fragments were cloned in Escherichia coli K-12 host using λgtWES·λB′ (λgtWES·λB, for short, in this paper) as a vector. Recombinant DNAs containing one or a few fragments of the mtDNA were transfected to CaCl2-treated E. coli, and the plaques containing specific recombinant phages were selected. DNA amplified in the recombinant phage λgt·mt was shown to contain the same restriction endonuclease cleavage sites as those found in the mtDNA. Present results permitted the DNA sequencing of any portion of the mitochondrial genome. © 1979.