Fructose-1, 6-bisphosphatase (EC 3.1.3.11, FBPase), a key regulatory enzyme int he gluconeogenic pathway, catalyzes the hydrolysis of fructose-1, 6- bisphosphate to fructose 6-phosphate the inorganic phosphate. Metabolic acidosis, hypoglycemia, and hepatomegaly have been reported by the deficiency of FBPase, which is also inherited as an autosomal recessive defect. We have isolated and partially characterized two closely related cDNA clones of FBPase from a chicken liver cDNA library in lambda gt 11. In the next phase, the genomic library of chicken liver constructed lambda dash will be screened using a radioactive cDNA probe. The authenticity of the isolated clone will be confirmed by comparing the deduced amino acid sequence with that obtained from direct sequencing of the enzyme using cyanogen bromide cleavage at methionine or by trypsin and lysyl endoproteinase digestion. This approach will facilitate the investigation of the structure of the gene (exons and introns), upstream promoter sequences and the transcription regulation of the gene. The tissue distribution of FBPase mRNA by RNA blot analysis will be assessed to explore molecular diversity within the family. The proposed investigation will provide undergraduate students with a variety of modern techniques used in recombinant DNA technology. This research will encourage them to pursue a research career in biomedical related areas of biochemistry or molecular biology.
| Bayse, Gladys S; Hammonds-Odie, Latanya P; Jackson, Kimberly M et al. (2013) Permeation of roxarsone and its metabolites increases caco-2 cell proliferation. Adv Biol Chem 3:389-396 |
