Abstract
Abstract: :
Purpose: Fovin is a novel gene product that is highly expressed in the retina. The purpose of this study was to determine the full-length sequence of the mouse fovin cDNA, to generate antibodies against fovin peptides and to ascertain the mouse tissues expressing the fovin protein. Methods: To identify the start of the coding region, 5’ RACE was performed using mouse retina cDNA and gene specific primers. The RACE products were cloned and analyzed by DNA sequencing and PCR. Two different peptides were deduced from the fovin cDNA sequence, synthesized and injected into two rabbits per peptide. The rabbits were bled to obtain the antibodies made against the fovin peptides. Western blots were run using mouse tissues and the fovin peptide antibodies. Results: Four RACE clones have been completely sequenced and differences are seen between them. A BLAST search revealed that the largest mouse fovin gene spans more than 45 kB on chromosome 11. Of the four fovin antibodies, one of them DEBRH1/80, recognizes a single band about 59 kDa in the mouse retina. DEBRH1/80 also recognizes several bands in the mouse brain and a single band in the mouse liver. Conclusions: Mapping of mouse fovin to chromosome 11 is not unexpected since human fovin maps to chromosome 17 and there is synteny between human chromosome 17 and mouse chromosome 11. Translation of the opening reading frame of the cDNA predicts that the mouse fovin protein is 59 kDa. One of the antibodies identifies a protein of about 59 kDa in the mouse retina indicating that this antibody recognizes fovin. Fovin isoforms are expressed in the mouse brain and liver. Alternative splicing may account for the differences seen between the 5’ RACE products and the bands detected by Western blots and may underlie a mechanism for functional diversity of this gene.
Keywords: retina • molecular biology • gene/expression