西伯利亚鲟肝脏中过氧化物酶体增殖物激活受体α、脂蛋白脂酶、肝脂酶基因全长 cDNA克隆与序列分析

摘要

本试验通过反转录聚合酶链式反应( RT-PCR)和cDNA末端快速扩增( RACE)技术克隆西伯利亚鲟( Acipenser baerii)肝脏中过氧化物酶体增殖物激活受体α( PPARα)、脂蛋白脂酶( LPL)、肝脂酶( HL )基因全长 cDNA 序列。结果显示：西伯利亚鲟肝脏中 PPARα基因( GenBank登录号KJ534588) cDNA全长1736 bp,包括158 bp的5’非翻译区、1401 bp的开放阅读框及177 bp的3’非翻译区；开放阅读框编码1个由466个氨基酸组成的蛋白质,预测其分子质量为52.3 ku,与其他物种的相似性为75.5%~82.4%。 LPL 基因( GenBank 登录号KJ720972) cDNA全长1760 bp,包括163 bp的5’非翻译区、1506 bp的开放阅读框及91 bp的3’非翻译区；开放阅读框编码1个由501个氨基酸组成的蛋白质,预测分子质量为57.1 ku,与其他物种的相似性为48.7%~67.0%。 HL基因( GenBank登录号KJ720973) cDNA全长1740 bp,包括124 bp的5’非翻译区、1500 bp的开放阅读框及116 bp的3’非翻译区；开放阅读框编码1个由499个氨基酸组成的蛋白质,预测分子质量为56.4 ku,与其他物种的相似性为55.4%~67.1%。通过构建系统发育进化树发现,西伯利亚鲟PPARα与高等脊椎动物的亲缘关系较近, LPL、HL与鱼类亲缘关系较近。本试验通过对西伯利亚鲟PPARα、HL、LPL基因同源性、系统进化地位的分析,为进一步研究西伯利亚鲟脂肪代谢调控机制奠定了分子生物学基础。%The full-length cDNAs of peroxisome proliferator-activated receptorα ( PPARα) , lipoprotein lipase ( LPL) and hepatic lipase ( HL) were cloned from Siberian sturgeon ( Acipenser baerii) liver using reverse transcription-phosphatase chain reaction ( RT-PCR) and rapid amplification of cDNA end ( RACE) methods in this study. The results showed as follows: the full-length cDNA of PPARα gene in liver of Siberian sturgeon (GenBank accession No. KJ534588) was 1 736 bp, including a 5’ untranslated region of 158 bp, an open reading frame ( ORF) of 1 401 bp and a 3’ untranslated region of 177 bp, the ORF encoding a protein of 466 amino acids with the theoretical molecular weight of 52.3 ku, and shared 75.5% to 82.4% similarity with other species. The full-length cDNA of LPL gene in liver of Siberian sturgeon ( GenBank accession No. KJ720972) was 1 760 bp, including a 5’ untranslated region of 163 bp, an ORF of 1 506 bp and a 3’ untranslated region of 91 bp, the ORF encoding a protein of 501 amino acids with the theoretical molecular weight of 57.1 ku, and shared 48.7% to 67. 0% similarity with other species. The full-length cDNA of HL gene in liver of Siberian sturgeon ( GenBank accession No. KJ720973) was 1 740 bp, including a 5’ untranslated region of 124 bp, an ORF of 1 500 bp and a 3’ untranslated region of 116 bp, the ORF encoding a protein of 499 amino acids with the theoretical molecular weight of 56.4 ku, and shared 55.4% to 67.1% similarity with other species. Phyloge-netic tree analysis revealed that PPARα gene of Siberian sturgeon had closer genetic relationship with higher vertebrates, but LPL and HL genes had closer genetic relationship with other fishes. In this study, the gene ho-mology and phylogenetic status of PPARα, HL, LPL from Siberian sturgeon are analyzed, and the results pro-vide a molecular biological basis for the further study of the regulatory mechanism of lipornetabolism in Siberi-an sturgeon.