Cloning and characterization of a cDNA specific for bovine retina

• Published in: Neuroscience research 1986-05, Vol.3 (4), p.300-310
• Main Authors: Nakagawa, Yuzo, Kuo, Che-Hui, Ishii, Kayoko, Shiosaka, Sadao, Tohyama, Masaya, Miki, Naomasa
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Biological and medical sciences; Cattle; Cell Line; Cloning, Molecular; DNA - isolation & purification; DNA Restriction Enzymes; Eye and associated structures. Visual pathways and centers. Vision; Fundamental and applied biological sciences. Psychology; Glioma - genetics; Humans; Mice; molecular cloning; neurnal retina; Nucleic Acid Hybridization; Poly A - isolation & purification; Rats; Retina - metabolism; retina specific cDNA; Retinoblastoma - genetics; RNA - isolation & purification; RNA, Messenger; Species Specificity; Vertebrates: nervous system and sense organs
• ISSN: 0168-0102; 1872-8111
• DOI: 10.1016/0168-0102(86)90022-2

A retina-specific cDNA clone (pCR18) was selected from a bovine retinal cDNA library and characterized. The clone pCR18 consisted of 905 base pairs and hybridized to the mRNA of about 12S from the bovine retina, but not that from the brain or liver. The nucleotide sequence revealed a long open reading frame which encodes a 147 amino acid polypeptide of about 15,700 Da. No significant sequence homology with the predicted protein was found in the protein sequence library of about 3500. Messenger RNA which hybridized to pCR18 translated a polypeptide of about 19,000 Da in a reticulocyte translation system. Southern blot analysis indicated that the bovine genome contains a single copy of this gene. Furthermore, RNA dot analysis showed that the poly(A) +RNA from the human retinoblastoma cell lines (Y79 and WERI) hybridized to pCR18, whose intensity was comparable to that of the bovine retina. In situ hybridization revealed that pCR18 was expressed mostly in some ganglion cells of the rat retina. The results suggest that cDNA clone (pCR18) encodes a protein specific for the retina and mRNA for pCR18 is mostly localized in the retinal ganglion cells and also expressed in the human retinoblastoma cells, although its function remains to be elucidated.