Cloning and sequencing of the alcohol dehydrogenase II gene from zymomonas mobilis

Journal Article · Mon Jun 01 00:00:00 EDT 1987 · J. Bacteriol.; (United States)

OSTI ID:6501556

Conway, T; Sewell, G W; Osman, Y A; Ingram, L O

The gene which encodes alcohol dehydrogenase II (adhB) from Zymomonas mobilis was cloned in Escherichia coli as a 1.4-kilobase DNA fragment by using a novel indicator plate which directly detects the expression of this activity by recombinant colonies. The DNA sequence for this clone contained an open reading frame encoding a polypeptide of 383 amino acids, with a molecular weight of 40,141. Although this protein exhibited very little homology with other known alcohol dehydrogenases, the predicted amino acid composition was in excellent agreement with that reported for the purified alcohol dehydrogenase II protein from Z. mobilis. In Z. mobilis, the adhB gene was transcribed from tandem promoters which were separated by 100 base pairs and ended with a transcriptional terminator (13-base-pair palindrome). In Escherichia coli, only one of the Z. mobilis promoters was used, despite apparent similarity to the enteric consensus promoter. The adhB gene was transcribed at low levels in E. coli from the P2 promoter of Z. mobilis but was expressed well in E. coli under control of the lac promoter (approximately 0.25% of the total cell protein).

• Research Organization: Univ. of Florida, Gainesville
• OSTI ID: 6501556
• Journal Information: J. Bacteriol.; (United States), Vol. 169:6
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
GENES
DNA SEQUENCING
DNA-CLONING
TRANSCRIPTION
OXIDOREDUCTASES
CHEMICAL COMPOSITION
ESCHERICHIA COLI
GENE REPRESSORS
MOLECULAR WEIGHT
ZYMOMONAS MOBILIS
BACTERIA
CLONING
ENZYMES
MICROORGANISMS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
PROTEINS
STRUCTURAL CHEMICAL ANALYSIS
550200* - Biochemistry