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Title: Molecular cloning of human ornithine aminotransferase mRNA

Abstract

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.

Authors:
; ; ; ; ; ;
Publication Date:
Research Org.:
National Institutes of Health (NIH), Bethesda, MD (United States)
OSTI Identifier:
5561717
Resource Type:
Journal Article
Journal Name:
Proc. Natl. Acad. Sci. U.S.A.; (United States)
Additional Journal Information:
Journal Volume: 83:5
Country of Publication:
United States
Language:
English
Subject:
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

Citation Formats

Inana, G, Totsuka, S, Redmond, M, Dougherty, T, Nagle, J, Shiono, T, Ohura, T. Kominami, E., and Katunuma, N. Molecular cloning of human ornithine aminotransferase mRNA. United States: N. p., 1986. Web. doi:10.1073/pnas.83.5.1203.
Inana, G, Totsuka, S, Redmond, M, Dougherty, T, Nagle, J, Shiono, T, Ohura, T. Kominami, E., & Katunuma, N. Molecular cloning of human ornithine aminotransferase mRNA. United States. https://doi.org/10.1073/pnas.83.5.1203
Inana, G, Totsuka, S, Redmond, M, Dougherty, T, Nagle, J, Shiono, T, Ohura, T. Kominami, E., and Katunuma, N. 1986. "Molecular cloning of human ornithine aminotransferase mRNA". United States. https://doi.org/10.1073/pnas.83.5.1203.
@article{osti_5561717,
title = {Molecular cloning of human ornithine aminotransferase mRNA},
author = {Inana, G and Totsuka, S and Redmond, M and Dougherty, T and Nagle, J and Shiono, T and Ohura, T. Kominami, E. and Katunuma, N},
abstractNote = {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.},
doi = {10.1073/pnas.83.5.1203},
url = {https://www.osti.gov/biblio/5561717}, journal = {Proc. Natl. Acad. Sci. U.S.A.; (United States)},
number = ,
volume = 83:5,
place = {United States},
year = {Sat Mar 01 00:00:00 EST 1986},
month = {Sat Mar 01 00:00:00 EST 1986}
}