Structure and expression of an alcohol dehydrogenase 1 gene from Pisum sativum (cv. "Greenfeast").

Three genomic clones for anaerobically inducible alcohol dehydrogenase (Adh) have been isolated from Pisum sativum cv. "Greenfeast" via cDNA cloning. One of these contains a complete gene, has exon sequences corresponding to one of the cDNA sequences and is likely to be an expressed gene. This gene has a structure similar to the Adh genes of maize, with introns in the same positions in the coding sequence but differing in their lengths and nucleotide sequences. At the nucleotide level the coding sequence is 75% homologous to both maize Adh1 and Adh2 and 80% homologous to the Adh gene from Arabidopsis, but has an extra coding triplet in exon 1 that is not found in the other plant Adh genes. The non-translated regions of all the gene transcripts are widely divergent between species. A short segment of the pea Adh promoter region (-290 to +57) was fused to a reporter gene and introduced into protoplasts of Nicotiana plumbaginifolia by electroporation. Transient expression of the introduced gene increased markedly when the transfected protoplasts were incubated under anaerobic conditions, showing that cis-acting regulatory signals necessary for anaerobic control of expression reside in the -290 to +57 segment. Sequence comparisons between this region and the corresponding regions of maize and Arabidopsis Adh genes have identified short sequences that may be involved in the anaerobic regulation of plant Adh genes.

[1]  P. Palukaitis,et al.  Replication of tobacco mosaic virus. VII. Further characterization of single- and double-stranded virus-related RNAs from TMV-infected plants. , 1983, Virology.

[2]  W. D. Benton,et al.  Screening lambdagt recombinant clones by hybridization to single plaques in situ. , 1977, Science.

[3]  W. Gerlach,et al.  Molecular Analysis of Ds Controlling Element Mutations at the Adh1 Locus of Maize , 1984, Science.

[4]  B. Barrell,et al.  Two related but differentially expressed potential membrane proteins encoded by the EcoRI Dhet region of Epstein-Barr virus B95-8 , 1985, Journal of virology.

[5]  E. Finnegan,et al.  Molecular analysis of the alcohol dehydrogenase 2 (Adh2) gene of maize. , 1984, Nucleic acids research.

[6]  W. Sofer,et al.  Alcohol dehydrogenase-negative mutants in Drosophila: defects at the structural locus? , 1976, Genetics.

[7]  B. Howard,et al.  Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells , 1982, Molecular and cellular biology.

[8]  D. Llewellyn,et al.  Maize Adh‐1 promoter sequences control anaerobic regulation: addition of upstream promoter elements from constitutive genes is necessary for expression in tobacco , 1987, The EMBO journal.

[9]  A. Hanson,et al.  Control of Lactate Dehydrogenase, Lactate Glycolysis, and α-Amylase by O2 Deficit in Barley Aleurone Layers , 1984 .

[10]  R. Crawford,et al.  A METABOLIC THEORY OF FLOODING TOLERANCE: THE SIGNIFICANCE OF ENZYME DISTRIBUTION AND BEHAVIOUR , 1971 .

[11]  Michael Freeling,et al.  The anaerobic proteins of maize , 1980, Cell.

[12]  T. Maniatis,et al.  Human β-interferon gene expression is regulated by an inducible enhancer element , 1985, Cell.

[13]  B. Hoffman,et al.  A simple and very efficient method for generating cDNA libraries. , 1983, Gene.

[14]  E. Chen,et al.  Supercoil sequencing: a fast method for sequencing plasmid DNA , 1985 .

[15]  F. Sanger,et al.  Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. , 1980, Journal of molecular biology.

[16]  J. Vieira,et al.  A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. , 1982, Gene.

[17]  I. Negrutiu Improved Conditions for Large-scale Culture, Mutagenesis, and Selection of Haploid Protoplasts of Nicotiana plumbaginifolia Viviani , 1981 .

[18]  J. Messing,et al.  Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. , 1983, Gene.

[19]  E. Chen,et al.  Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. , 1985, DNA.