猪PNLIPRP2基因的分离、分子特征以及组织表达谱研究  

Isolation,Molecular Characterization and Tissue-specific Expression Analysis of the Porcine PNLIPRP2 Gene

作　　者：程蕾[1,2] 符亚原[3] 刘晓华 王定发 王肆玖 左波[1] 熊远著[1] 

机构地区：[1]华中农业大学动物科技学院,武汉430070 [2]武汉市畜牧兽医研究所,武汉430065 [3]黑龙江八一农垦大学动物科技学院,大庆163319

出　　处：《畜牧兽医学报》2009年第9期1283-1289,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金："863"国家高技术研究发展计划(2006AA10Z1D6);"973"国家重大基础研究项目(2006CB102102)

摘　　要：本研究旨在分离鉴定猪胰脂肪酶相关蛋白2(Pancreatic lipase-related protein 2,PNLIPRP2)基因,并对其分子特征及在主要组织中的表达分布进行探讨。利用电子克隆和RT-PCR技术克隆出猪PNLIPRP2基因编码区(CDS)全长,并将序列提交到GenBank;采用GENSCAN等软件和NCBI等在线资源对其及其预测编码蛋白序列进行生物信息学和比较基因组学分析;同时利用半定量PCR技术分析该基因在2月龄大白猪12种组织中的表达情况。结果表明,猪PNLIPRP2基因CDS全长1 416 bp(EU715062),编码471个氨基酸,含有1个保守的甘油三酯脂酶抑肽酶PLAT结构域(PLAT-PL),在进化上与人、黑猩猩PNLIPRP2的遗传距离较近。进一步分析显示,该基因定位于猪14号染色体,含有12个外显子和11个内含子。另外,猪PNLIPRP2在小肠中表达量最高,脑、脂肪中次之,骨髓、胃中表达量较低,在其它组织中几乎不表达。本研究对猪PNLIPRP2基因CDS进行了克隆,初步分析了该基因的结构与表达特征,结果提示,猪PNLIPRP2可能是与脂肪沉积和免疫调节密切相关的重要候选基因。The aim of the study is to clone the full-length coding region of the porcine PNLIPRP2（pancreatic lipase-related protein 2） gene,analyze the molecular characteristic,and investigate its expression in the major tissues.The CDS of porcine PNLIPRP2 gene was cloned using the in Silico Cloning and RT-PCR methods.The sequence was submitted to the GenBank.Many softwares（e.g.,GENSCAN） and online resources（e.g.,NCBI） were used for the analysis of bioinformatics and comparative genomics.The expression of PNLIPRP2 gene was investigated by RT-PCR in 12 different tissues of 2-month-old Large White pig.The full-length CDS of the porcine PNLIPRP2 is 1 416 bp（EU715062） encoding a protein of 471 amino acids which contains a conserved PLAT domain of pancreatic triglyceride lipase（PLAT-PL）.Porcine PNLIPRP2 is evolutionary more near to human and chimpanzee than other species.Further bioinformatics analysis showed that porcine PNLIPRP2 gene was located on porcine chromosome 14 and contained 12 exons and 11 introns.In addition,porcine PNLIPRP2 mRNA was the most abundant in intestine,followed by the mordrate level in brain and fat,the lowest level in bone marrow and stomach,and almost no expression in other tissues.The CDS full-length of the porcine PNLIPRP2 gene was cloned.The molecular characteristic and its expression patterns in different tissues were also described.The results indicated that porcine PNLIPRP2 gene may be an important candidate gene in regarding to the fatty deposition and the immunity.

关 键 词：猪 胰脂肪酶相关蛋白2基因 序列分析 半定量RT-PCR 

分 类 号：S828[农业科学—畜牧学] S813.3[农业科学—畜牧兽医]