A Novel Human Lipid Binding Protein Coding Gene: PERF15, Sequence and Cloning

Introduction: PERF15 is a testicular germ-cell specific fatty-acid binding protein(FABP) isolated from mammals, originally from rats. It encodes one of the mostabundant proteins of rat spermatozoa localized in the perinuclear theca. Northern blotanalysis has demonstrated that rat PERF15 mRNA is exclusively transcribed duringmeiosis and post-meiosis. In this study, we cloned and sequenced human PERF15gene.Materials and Methods: According to the open reading frame of automatedcomputational analysis of Homo sapiens similar to testis fatty acid binding proteinnine, two specific Primers were designed to amplify human PERF15 gene. To confirmthe identity of the amplified gene, PCR products of PERF15 were cloned intoappropriate plasmid vectors followed by sequencing of the inserts.Results: A unique band of ~3kb was obtained after PCR amplification. Restrictionenzyme digestion using PvuII confirmed that the fragment was related to PERF15.Gene alignment, direct sequencing and application of specific primers to the geneshowed 100% similarity between this gene and the computational data by gelextraction of the ~3 kb band. The human PERF15 gene contained four exons and threeintrons. Exons one, two, three and four, respectively, coded for 24, 57, 34 and 17amino acids. The existing three introns were composed of 2113, 461, and 168nucleotides.Conclusion: In spite of the homology between exonic regions and exon-intronboundaries of human PERF15 gene and that of animals, human PERF15 gene isdifferent in size and sequence from corresponding introns in rat and murine PERF15.