Cloning and SequenceAnalysis of a Novel Gene EncodingA Testis Lipid Binding Protein(PERF15) in Human Testis


Pouresmaeili F *, Tehrani M.K ,

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PERF15 is a testicular germ cell specific fatty acid-binding protein (FABP) isolated from mammals, originally, rat. It encodes for one of the most abundant proteins of the rat spermatozoa localized in perinuclear theca. It is a small protein of 15 060 molecular weight. Northern blot analysis demonstrated that the rat PERF15 mRNA is transcribed in meiotic and post meiotic cells, exclusively. In this study, we have cloned the human PERF15 gene. For this purpose, we have designed two specific primers to amplify the human PERF15 gene according to the open reading frame of automated computational analysis of Homo sapiens similar to testis fatty acid binding protein 9 with accession #XM_378035. After performance the PCR, a unique band of ~3kb was obtained. Restriction digest using PvuII restriction enzyme, confirmed that the fragment is Abstract of the 8th Royan International Twin Congress, Tehran, Iran, 5-7 September 2007 20 Yakhteh Medical Journal, Vol 9, Sup 1, Summer 2007 related to the same gene. Then we gel extracted the ~3kb band. We sequenced this fragment by direct sequencing using Automatic sequencer (version 3130XL Genetic Analyzer, ABI Applied Biosystem) and 2 specific primers to the gene. Alignment showed 100 % similarity between our gene sequence and the mentioned computational data. The human PERF15 gene contains four exons and three introns. Exon 1 codes for 24 amino acids, exon 2 codes for 57 amino acids, exon 3 codes for 34 amino acids and exon 4 codes for 17 amino acids, respectively. The existing three introns are composed of 2113, 461, and 168 nucleotides. In spite of the homology between exonic regions and exonintron boundaries of the human PERF15 gene and the animal’s (rat& mouse PERF15 genes), the human PERF15 gene is different in size and sequence of the corresponding introns with rat and mouse PERF15.