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Isolation and characterization of a ColE1 plasmid containing the entireBiogene cluster ofEscherichia coliK12

✍ Scribed by Gerald Cohen; Zehava Zimmer; Raya Gurevich; Saul Yankofsky

Publisher
Springer
Year
1978
Tongue
English
Weight
842 KB
Volume
166
Category
Article
ISSN
1617-4615

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✦ Synopsis

ColE1amp plasmids carrying the entire bio gene cluster were constructed in vitro using ColE1amp as the cloning vehicle and a lambda transducing phage, lambdaatt2, as the source of bio DNA. Restriction endonuclease EcoRI digests of ColE1amp and lambdaatt2 DNA were joined by polynucleotide ligase and plasmids bearing the entire bio gene cluster were selected, after transformation, in bio deletion strains of E. coli. Recombinant DNA molecules contained one ColE1amp fragment (7.4 X 10(6) daltons) and one lambdaatt2 DNA fragment (5.4 X 10(6) daltons). Clones carrying ColE1 amp-bio plasmids produce elevated levels of biotin and biotin synthetase activity.

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