Farkhondeh Pouresmaeili, Tahereh Khalili, Mahmood Jeddi Tehrani, Mojgan Bandehpour, Jamileh Ghasemi, Fatemeh Salehi-Niya
Department of Genetics, Faculty of Medicine, Shaheed Beheshti University of Medical Sciences, Tehran, Iran ; Fertility-Infertility Health Research Center (IRHRC), Taleghani Hospital, Shaheed Beheshti University of Medical Sciences, Tehran, Iran.
J Reprod Infertil. 2009 Oct;10(3):199-205.
PERF15 is a testicular germ-cell specific fatty-acid binding protein (FABP) isolated from mammals, originally from rats. It encodes one of the most abundant proteins of rat spermatozoa localized in the perinuclear theca. Northern blot analysis has demonstrated that rat PERF15 mRNA is exclusively transcribed during meiosis and post-meiosis. In this study, we cloned and sequenced human PERF15 gene.
According to the open reading frame of automated computational analysis of Homo sapiens similar to testis fatty acid binding protein nine, two specific Primers were designed to amplify human PERF15 gene. To confirm the identity of the amplified gene, PCR products of PERF15 were cloned into appropriate plasmid vectors followed by sequencing of the inserts.
A unique band of ∼3kb was obtained after PCR amplification. Restriction enzyme digestion using PvuII confirmed that the fragment was related to PERF15. Gene alignment, direct sequencing and application of specific primers to the gene showed 100% similarity between this gene and the computational data by gel extraction of the ∼3 kb band. The human PERF15 gene contained four exons and three introns. Exons one, two, three and four, respectively, coded for 24, 57, 34 and 17 amino acids. The existing three introns were composed of 2113, 461, and 168 nucleotides.
In spite of the homology between exonic regions and exon-intron boundaries of human PERF15 gene and that of animals, human PERF15 gene is different in size and sequence from corresponding introns in rat and murine PERF15.
PERF15是一种从哺乳动物（最初是大鼠）中分离出的睾丸生殖细胞特异性脂肪酸结合蛋白（FABP）。它编码大鼠精子中最丰富的蛋白质之一，定位于核周膜。Northern印迹分析表明，大鼠PERF15 mRNA仅在减数分裂和减数分裂后转录。在本研究中，我们克隆并测序了人类PERF15基因。
根据对与睾丸脂肪酸结合蛋白9相似的智人的自动计算分析的开放阅读框，设计了两个特异性引物来扩增人类PERF15基因。为了确认扩增基因的身份，将PERF15的PCR产物克隆到合适的质粒载体中，然后对插入片段进行测序。
PCR扩增后获得了一条约3kb的独特条带。使用PvuII进行的限制性酶切证实该片段与PERF15相关。基因比对、直接测序以及对该基因应用特异性引物显示，通过凝胶提取约3kb条带，该基因与计算数据之间具有100%的相似性。人类PERF15基因包含四个外显子和三个内含子。外显子一、二、三、四分别编码24、57、34和17个氨基酸。现有的三个内含子分别由2113、461和168个核苷酸组成。
尽管人类PERF15基因的外显子区域和外显子 - 内含子边界与动物的具有同源性，但人类PERF15基因在大小和序列上与大鼠和小鼠PERF15的相应内含子不同。
