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Expression of the cloned uvrB gene of Escherichia coli: mode of transcription and orientation

Journal Article · · J. Bacteriol.; (United States)
OSTI ID:5334120
; ;
The Escherichia coli uvrB gene, located on a 1.5-megadalton EcoRI fragment F, derived from transducing phage lambda b2att/sup 2/ (lambda b2cI857intam6..delta..(bioAB)bioFCD+uvrB+), has been cloned in the unique EcoRI site of several relaxed plasmids, i.e., pMB9, pBR322, and pBH20 (=pBR322, including the lac regulatory elements). Expression of the uvrB gene, both on pMB9 and on pBH20, occurs only when fragment F has one particular orientation. Cloning of this fragment on pBR322 in either orientation does not allow expression of the uvrB gene. Transcription of this gene on pNP5 (=pMB9 uvrB) is shown to be dependent on a pMB9 promotor that is located on a 0.22-megadalton EcoRI-HindIII fragment. Using plasmid pBH20 as a vector, we could demonstrate that expression of the uvrB gene is under control of the lac promotor-operator region. From deoxyribonucleic acid-deoxyribonucleic acid hybridization experiments with lambda pgal8 deoxyribonucleic acid and restriction fragments of pNP5 deoxyribonucleic acid it could be shown that the uvrB gene is transcribed clockwise on the chromosome.
Research Organization:
Univ. of Leiden, Netherlands
OSTI ID:
5334120
Journal Information:
J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 139:1; ISSN JOBAA
Country of Publication:
United States
Language:
English

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Related Subjects

550200 -- Biochemistry
550400 -- Genetics
560112* -- Radiation Effects on Biochemicals-- In Microorganisms-- (-1987)
560131 -- Radiation Effects on Microorganisms-- Basic Studies-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
BACTERIA
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CELL CONSTITUENTS
CLONING
DNA
ELECTROMAGNETIC RADIATION
ESCHERICHIA COLI
GENE MUTATIONS
GENES
KINETICS
MICROORGANISMS
MUTATIONS
NUCLEIC ACIDS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
ORIENTATION
PLASMIDS
PROTEINS
RADIATION EFFECTS
RADIATIONS
RADIOSENSITIVITY
RADIOSENSITIVITY EFFECTS
REACTION KINETICS
RECOVERY
REPAIR
TRANSCRIPTION
ULTRAVIOLET RADIATION