| 草鱼碱性氨基酸转运载体y+LAT2 cDNA基因克隆与表达研究 |
| Molecular Cloning and Expression Analysis of cationic amino acid transporter y+LAT2 from Grass Carp |
| 投稿时间：2013-01-13 修订日期：2013-03-10 |
| DOI： |
| 中文关键词:草鱼 碱性氨基酸转运载体 y+LAT2 基因克隆 表达研究 |
| 英文关键词:Grass carp Cationic amino acid transporter y+LAT2 gene cloning expression analysis |
| 基金项目: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） |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| The cDNA sequence encoding y+LAT2 gene was cloned and sequenced from grass carp (Ctenopharyngodon idellus ) by RT – PCR and RACE. The tissue distribution and expression regulation by fasting of y+LAT2 mRNA were analyzed using real – time PCR. The results showed that the cDNA (1849 bp) encodingy+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 amphibian. The phylogenetic tree of amino acid sequence built from y+LAT2 homology of grass crap 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. |
| 杨吉轩,谭青松,朱文欢,陈 忱.2013.草鱼碱性氨基酸转运载体y+LAT2 cDNA基因克隆与表达研究[J].水生态学杂志,34(3):54-61. |
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