Molecular Cloning of ELOVL4 Gene from Cynomolgus Monkey (Macaca Fascicularis)

Abstract

Abstract: : Purpose: To clone the monkey homologue of the ELOVL4, the gene responsible for autosomal dominant Stargardt-like macular dystrophy (STGD3) and to determine its expression profile in various tissues and protein distribution in monkey retina. Methods: PCR of genomic DNA and 5’/3’-rapid amplification of cDNA ends were carried out to determine the complete sequence of the monkey ELOVL4 cDNA and the gene. ELOVL4 mRNA expressions in various tissues were measured by real-time quantitative PCR using various tissues and immunohistochemical analysis was performed using anti-human ELOVL4 peptide polyclonal antibody to localize ELOVL4 protein in monkey retina. Results: Sequence analysis of monkey ELOVL4 revealed similar gene structure to human. The cloned full-length cDNA encoded 314 amino acids, same length as human and two amino acids shorter than mouse. The monkey ELOVL4 also preserved characteristics typical to super family of ELO, enzymes involved in the metabolism of membrane-bound fatty acid elongation. The monkey ELOVL4 was highly expressed in restricted tissue-specific fashion in skin (90% of retina) and in thymus (111% of retina). Among other tissues, significant expression was observed only in brain at the level less than 9% of the retina. Immunohistochemical study revealed that the ELOVL4 protein is expressed in rod and cone photoreceptors of monkey retina. Conclusion: ELOVL4 was cloned from Cynomolgus monkey (Macaca Fascicularis). Gene sequence and structure of ELOVL4 was similar to human and mouse homologues. Quantitative-PCR experiments revealed the ELOVL4 mRNA expression not only in retina but also in thymus and skin. Immunohistochemical studies localize the expression of ELOVL4 protein to rod and cone photoreceptors.

Keywords: 308 age-related macular degeneration • 476 molecular biology • 417 gene/expression