Isolation and Characterization of theDrosophilaOrnithine Decarboxylase Locus: Evidence for the Presence of Two Transcribed ODC Genes in theDrosophilaGenome
- 1 July 1993
- journal article
- Published by Mary Ann Liebert Inc in DNA and Cell Biology
- Vol. 12 (6) , 499-508
- https://doi.org/10.1089/dna.1993.12.499
Abstract
The polymerase chain reaction (PCR) was used to isolate two Drosophila ornithine decarboxylase (ODC) genes. Two mixtures of degenerate oligonucleotides corresponding to peptides that are fully conserved among ODCs from widely diverged species were used as opposing primers in the PCR with cDNA or genomic DNA as templates. Sequence analysis of the resulting DNA products confirmed their identity as ODC fragments. The genomic PCR product was then used as a probe for screening a Drosophila genomic library, resulting in the isolation of genomic clones representing two distinct ODC genes (dODC1 and dODC2). Sequence analysis of both genes demonstrated that although varying at their coding and noncoding regions, their overall structure is extremely similar containing 6 exons and 5 short introns. Southern blot and sequence analyses revealed that the two ODC genes are arranged in a tandem head-to-tail configuration. Both ODC genes were assigned by in situ hybridization analysis to position 44A on the right arm of the second chromosome. The isolation of cDNA clones corresponding to these two ODC genes demonstrated that both are transcribed in the adult fly. We hope that the isolation of genomic and cDNA clones of Drosophila ODC will permit the investigation of the expression of ODC during Drosophila development and the role of polyamines in this process.Keywords
This publication has 19 references indexed in Scilit:
- The Drosophila FGF-R homolog is expressed in the embryonic tracheal system and appears to be required for directed tracheal cell extension.Genes & Development, 1991
- Characterization of sequences involved in mediating degradation of ornithine decarboxylase in cells and in reticulocyte lysateEuropean Journal of Biochemistry, 1991
- Characterization and Sequence Analysis of the Human Ornithine Decarboxylase GeneDNA, 1989
- Prevention of Rapid Intracellular Degradation of ODC by a Carboxyl-Terminal TruncationScience, 1989
- Use of the DNA polymerase chain reaction for homology probing: isolation of partial cDNA or genomic clones encoding the iron-sulfur protein of succinate dehydrogenase from several species.Proceedings of the National Academy of Sciences, 1989
- The molecular genetics of embryonic pattern formation in DrosophilaNature, 1988
- Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA PolymeraseScience, 1988
- Enzymatic Amplification of β-Globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell AnemiaScience, 1985
- Isolation of biologically active ribonucleic acid from sources enriched in ribonucleaseBiochemistry, 1979
- Preparation, molecular weight, base composition, and secondary structure of giant nuclear ribonucleic acidBiochemistry, 1973