Molecular cloning of human ornithine aminotransferase mRNA

Journal Article · Sat Mar 01 00:00:00 EST 1986 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.83.5.1203· OSTI ID:5561717

Inana, G; Totsuka, S; Redmond, M; Dougherty, T; Nagle, J; Shiono, T; Katunuma, N

The isolation and characterization of a cDNA clone for the mRNA of human ornithine aminotransferase (OATase; ornithine-oxo-acid aminotransferase; L-ornithine:2-oxo-acid aminotransferase, EC 2.6.1.13), a nonabundant mitochondrial matrix enzyme that is severely deficient in a hereditary chorioretinal degenerative disease (gyrate atrophy), is described. Human liver, retina, and retinoblastoma (Y79) mRNAs were prepared and tested for the OATase mRNA content by in vitro translation, immunoprecipitation, and NaDodSO/sub 4//PAGE. The retinoblastoma cells were found to be expressing this enzyme at a relatively high level. The primary translation product of the OATase mRNA is larger than the pure OATase protein on NaDodSO/sub 4//PAGE. lambdagt11 cDNA libraries were prepared from the human mRNAs, and the recombinant clones were immunoscreened as plaques with two different preparations of rabbit anti-human OATase antibodies. The amino acid sequences of seven tryptic peptides (115 amino acid residues) of the pure human OATase were obtained by microsequencing. When the tryptic peptide and cDNA-derived amino acid sequences were compared, homologies in 111 of 115 residues, including a match of 20 consecutive residues, were observed. An RNA blot hybridization of /sup 32/P-labeled OATase cDNA to normal human retina and retinoblastoma mRNAs demonstrated an OATase mRNA species of approx. = 2.2 kilobases.

• Research Organization: National Institutes of Health (NIH), Bethesda, MD (United States)
• OSTI ID: 5561717
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 83:5
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
AMINOTRANSFERASES
BIOCHEMISTRY
MESSENGER-RNA
DNA-CLONING
AMINO ACID SEQUENCE
ELECTROPHORESIS
LIVER
MAN
PHOSPHORUS 32
RECOMBINANT DNA
RETINA
SULFUR 35
TUMOR CELLS
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
BODY AREAS
CHEMISTRY
CLONING
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
DNA
ENZYMES
EVEN-ODD NUCLEI
EYES
FACE
GLANDS
HEAD
ISOTOPES
LIGHT NUCLEI
MAMMALS
MOLECULAR STRUCTURE
NITROGEN TRANSFERASES
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS ISOTOPES
PRIMATES
RADIOISOTOPES
RNA
SENSE ORGANS
SULFUR ISOTOPES
TRANSFERASES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques