CAJ | 학술논문

本文构建了鲤鱼肝胰脏cDNA文库,共获得了1016条有效的表达序列标签.拼接组装成115个contigs和282个single-tons.其中215个拼接序列在GenBank公共数据库中寻找到相对应的基因.对它们进行功能性分类和比较分析为鲤鱼肝胰脏的研究提供了基因表达信息的基础.文库中1016条表达序列标签有11条代表了鲤鱼肝基本型脂肪酸结合蛋白(Lb-FABP).通过序列比较我们获得了两个具有相同开放阅读框长度的Lb-Fabp cDNAs.开放阅读框全长381bp,编码126个氨基酸.半定量RT-PCR结合Southern blot技术研究了L6-Fabp mRNA在成鱼不同组织以及早期发育不同时期的表达图式.结果表明,Lb-FabP mRNA在肝胰脏、中肠和后肠中表达量较高.同时在精巢和皮肤中有低水平的表达.脑、肌肉、卵巢、肾脏、脾脏、鳃和心脏等组织中其表达量更低.而在脂肪和前肠中则没有检测到Lb-Fabp mRNA表达.Lb-Fabp mRNA最早在胚体形成期检测到有低水平表达,随后的发育阶段中表达量逐渐升高.鲤鱼Lb-Fabp基因的表达图式提示在肝脏和肠等器官开始发育后,它可能在脂肪代谢中具有重要作用.
본문구건료리어간이장cDNA문고,공획득료1016조유효적표체서렬표첨.병접조장성115개contigs화282개single-tons.기중215개병접서렬재GenBank공공수거고중심조도상대응적기인.대타문진행공능성분류화비교분석위리어간이장적연구제공료기인표체신식적기출.문고중1016조표체서렬표첨유11조대표료리어간기본형지방산결합단백(Lb-FABP).통과서렬비교아문획득료량개구유상동개방열독광장도적Lb-Fabp cDNAs.개방열독광전장381bp,편마126개안기산.반정량RT-PCR결합Southern blot기술연구료L6-Fabp mRNA재성어불동조직이급조기발육불동시기적표체도식.결과표명,Lb-FabP mRNA재간이장、중장화후장중표체량교고.동시재정소화피부중유저수평적표체.뇌、기육、란소、신장、비장、새화심장등조직중기표체량경저.이재지방화전장중칙몰유검측도Lb-Fabp mRNA표체.Lb-Fabp mRNA최조재배체형성기검측도유저수평표체,수후적발육계단중표체량축점승고.리어Lb-Fabp기인적표체도식제시재간장화장등기관개시발육후,타가능재지방대사중구유중요작용.
A representative cDNA library obtained from hepatopanereas of common carp (Cyprinus carpio) was constructed, and 1016 expressed sequence tags were generated assembling into 115 contiguous sequences and 282 singletons. 215 (54.15%) of the assembled sequences were annotated based upon matching known genes in the GenBank public Databas-es. Functional categorization and comparative analyses of the sequence provided gene identities, expression information and a solid base for further research on common carp hepatopancreas. High expression of liver basic fatty acid binding protein (Lb-Fabp) (11/1016 ESTs) in the library was detected. Two types of Lb-Fabpa and Lb-Fabpb had an open reading frame of 381 bp encoding 126 amino acids, mRNA expression of Lb-Fabps in embryos, larva and adult common carp tissues were determined using semi-quantitative RT-PCR followed by Southern Blot analysis. Lb-Fabp mRNA was expressed with high levels in hepatopancreas, middle intestine and posterior intestine. Low levels of Lb-Fabp mRNA expression were observed in testis and skin, while much lower levels in muscle, brain, ovary, kidney, spleen, gill and heart. No Lb-Fabp mRNA expression was observed in fat and anterior intestine. Lb-Fabp mRNA was first observed in neurula stage, and expressed higher in the following stages. The expression pattern of Lb-Fabp mRNA suggests that common carp Lb-FABPs play an im-portant role in lipid metabolism after the morphogenesis of hepatopancreas and intestine.