草鱼碱性氨基酸转运载体y^＋ LAT2 cDNA基因克隆与表达  

Molecular Cloning and Expression Analysis of Cationic Amino Acid Transporter y^+ LAT2 from Ctenopharyngodon idellus

作　　者：杨吉轩[1] 谭青松[1] 朱文欢[1] 陈忱[1] 

机构地区：[1]华中农业大学水产学院农业部淡水生物繁育重点实验室,湖北武汉430070

出　　处：《水生态学杂志》2013年第3期54-61,共8页Journal of Hydroecology

基　　金：华中农业大学自主科技创新基金(2009QC022);公益性行业(农业)科研专项经费(201003020)

摘　　要：利用逆转录聚合酶链式反应(RT-PCR)和cDNA末端快速扩增(RACE)方法,克隆y+LAT2基因cDNA序列,并利用实时荧光定量PCR探讨y+LAT2在草鱼(Ctenopharyngodon idellus)各组织中的表达情况以及饥饿对其mRNA的影响。结果表明,获得的y+LAT2基因的cDNA序列片段为1849bp,含1371bp的核心序列,编码456个氨基酸。预测草鱼氨基酸序列与斑马鱼(Danio rerio)的同源性高达96.5%,与哺乳动物和两栖动物的同源性在78%～83%,构建的系统进化树与传统形态学分类一致。在草鱼的肌肉、脑、鳃、心、肾脏、肝、后肠、中肠、前肠、脾脏组织均检测到y+LAT2基因的表达。草鱼脾脏、肾脏以及前肠、中肠的y+LAT2 mRNA表达水平对禁食的反应不同,在短期饥饿(14d)期间,其y+LAT2 mRNA表达水平均呈下降趋势。草鱼碱性氨基酸转运载体y+LAT2基因全长cDNA序列的获得,有利于进一步探讨鱼类氨基酸的吸收转运机制。The cDNA sequence encoding y＋LAT2 gene was cloned and sequenced from grass carp （Ctenopharyngodon idellus） by RT-PCR and RACE method. The tissue distribution and expression regulation by fasting of y＋LAT2 mRNA were analyzed using real-time PCR.The results showed that the cDNA （1 849 bp） encoding y＋LAT2 contained a core sequence （1 371 bp） encoding 456 amino acids. Amino acid sequence analysis showed that sequence homology of C. idellus was 96.5% identity with the sequence of Danio rerio and between 78% and 83% with sequences of mammalian and amphibian. The phylogenetic tree of amino acid sequence built from y＋LAT2 homology of grass carp and other animals was consistent with that of the traditional morphology results.The y＋LAT2 mRNA was detected in muscle, brain, gill, heart, kidney, liver, hindgut, midgut, foregut and spleen. The expression patterns of y＋LAT2 in spleen, kidney and foregut, midgut were different and y＋LAT2 mRNA expression after fasting （14 days） showed a decreasing trend. The research laid a foundation for further study on the metabolism of fish amino acid absorption and transport.

关 键 词：草鱼 碱性氨基酸转运载体 y+LAT2 基因克隆 

分 类 号：Q343.1[生物学—遗传学]