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Molecular cloning and characterization of the alkB gene of Escherichia coli

✍ Scribed by Kataoka, Hiroko ;Sekiguchi, Mutsuo

Publisher: Springer
Year: 1985
Tongue: English
Weight: 825 KB
Volume: 198
Category: Article
ISSN: 0026-8925
DOI: 10.1007/bf00383004

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✦ Synopsis

Using methods of in vitro recombination we constructed hybrid plasmids that can suppress the increased methylmethane sulfonate sensitivity caused by alkB mutation. Since the cloned DNA fragment was mapped at 47 rain on the Escherichia coli K12 genetic map, an area where the alkB gene is located, we concluded that the cloned DNA fragment contains the alkB gene itself but not other genes that suppress alkB mutation. Specific labeling of plasmid-encoded proteins by the maxicell method revealed that the alkB codes for a polypeptide with a molecular weight of about 27,000. Introduction of a small deletion into the alkB region of the bacterial chromosome resulted in inactivation of both the alkB and ada genes, thereby suggesting that the two genes are adjacent on the E. coli chromosome.

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