A suitable method for construction and cloning hybrid plasmids containing EcoRI-fragments of E. coli genome. |
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Authors: | J. I. Kozlov N. A. Kalinina L. V. Gening B. A. Rebentish A. Y. Strongin V. G. Bogush V. G. Debabov |
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Affiliation: | (1) Institute of Genetics and Selection of Industrial Microorganisms, P.O. Box 825, 113545 Moscow, USSR |
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Abstract: | Summary A convenient procedure for the isolation of specificEcoRI-fragments ofE. coli genome and their amplification on Km-resistance plasmid vector CK 11 is described. The hybrid molecules were constructedin vitro usingEcoRI-digestion, followed by ligation. Then appropriatedE. coli strain was transformed with ligated DNA mixture and hybrid plasmids CK 11-arg+, CK 11-his+, CK 11-thr+ and CK 11-leu+ containing loci ofE. coli genome were selected by molecular cloning. The hybrid plasmids obtained consisted of oneEcoRI-fragment of initial plasmid CK 11 and one respective specific portion ofE. coli genome. |
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