摘要
为了研究鳜(Siniperca chuatsi)脂蛋白脂酶(LPL)、肝脂酶(HL)基因结构与功能关系,首先采用RT-PCR及RACE法,克隆得到鳜肝脏LPL与HL基因cDNA全序列。鳜肝脏LPL基因cDNA全长为2089bp,其中开放阅读框(ORF)为1548bp,编码515个氨基酸。鳜HL基因cDNA全长为1964bp,其中ORF为1494bp,编码497个氨基酸。进化树分析显示,鳜LPL与HL基因与其他脊椎动物的LPL、HL基因各自聚集成簇。对鳜基因组进行PCR获得鳜LPL与HL全基因组DNA序列,与其他脊椎动物基因结构相似,鳜LPL基因由10个外显子和9个内含子组成,全长7392bp;鳜HL基因由9个外显子和8个内含子组成,全长8837bp。应用Genome Walker方法在鳜克隆得到一段长为1071bp的LPL和一段长为2173bp的HL基因5′侧翼区序列,并利用相关软件预测其中具有多个保守的顺式调控元件。组织表达研究显示与易患动脉粥样硬化的人类LPL不同,鳜LPL基因在所有被检测组织中均有表达:在脂肪组织中大量表达,其次是肝脏、脑、肠道、肌肉,在脾中表达量最低;而鳜HL基因的表达则与人类相似,只在肝脏中表达。本研究将为深入探讨机体脂质代谢调节机制以及LPL、HL在防治动脉粥样硬化中的作用奠定良好基础。
In order to study the genomic structure and functional of Siniperca chuatsi lipoprotein lipase （LPL） and hepatic lipase （HL） genes, at first, two full-length cDNA clones of LPL and HL were isolated from the liver of Siniperca chuatsi by RT-PCR and RACE. The LPL cDNA was 2 089 bp in length, and contains a 1 548 bp open reading frame （ORF） encoding 515 amino acids. The HL cDNA was 1 964 bp in length, and contains a 1 494 bp ORF encoding 497 amino acids. Phylogenetic analysis showed that Siniperca chuatsi LPL and other vertebrate LPL are grouped in one cluster, as well as all the HL sequences grouped in the other cluster. LPL and HL genomic sequences were identified in the Siniperca chuatsi genome and displayed the same genomic structure as other vertebrates （LPL： 10 exons and 9 introns; HL： 9 exons and 8 introns） spanning 7.4 kb and 8.8 kb, respectively. Using genome walker method, the LPL and HL 5＇ -flanking regions were obtained in the Siniperca chuatsi which spanned 1 071 bp and 2 173 bp, respectively, and several potential regulatory elements were identified in the promoter regions. The mRNA expression of Siniperca chuatsi LPL gene was detected in all the examined tissues： highest in adipose tissue, followed by liver, brain, intestine, muscle, and lowest in spleen. But the Siniperca chuatsi HL gene was found to be exclusively expressed only in liver. These studies will lay a foundation in the regulation of gene expression in lipid metabolism and the prevention from the development of atherosclerosis.
出处
《中国水产科学》
CAS
CSCD
北大核心
2009年第4期506-517,共12页
Journal of Fishery Sciences of China
基金
广东省海洋渔业科技推广专项资助项目(A200899D02)
广东省自然科学基金资助项目(031886
980702)
