| 摘要: |
| 采用 RT-PCR 和 RACE 技术克隆鲫鱼 slc7A8 基因的全长 cDNA 序列, 对基因序列用生物软件对基因进行生物信息分析, 用 Real-time PCR 方法检测基因在组织中的 mRNA 表达丰度。结果表明, 鲫鱼 slc7A8cDNA 全长为 2709bp, 包含 195bp 的 5′UCR 序列, 921bp 的 3′UCR 序列, 1593bp 开放阅读框, 编码 530 个氨基酸序列; 鲫鱼与斑马鱼基因同源性和编码氨基酸同源性分别为 88.9%和95.5%, 而与其他动物分别在 70.1%—74.8%和 80.6%—82.1%之间; 预测编码蛋白的分子量为57719.84, 等电点为 4.8, 对其结构预测显示该蛋白具有与哺乳动物十分相似的 12 个螺旋跨膜结构, 跨膜区氨基酸高度保守, 不同动物间的同源性都在 92.2%以上; Real-time PCR 检测鲫鱼组织中的mRNA 表达丰度高低顺序为前肠、中肠、脑、后肠、肌肉、肾、鳃、心、肝脏。
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| 关键词: 鲫鱼, slc7A8 cDNA, 分子特征, slc7A8 mRNA 表达丰度 |
| DOI：10.11693/hyhz201006021021 |
| 分类号: |
| 基金项目:国家自然科学基金资助项目, 31001114 号; 湖南省高等学校科学研究项目(优秀青年项目), 09B011 号; 湖南省高校科技创新团队支持计划资助, 2008—2010。 |
附件 |
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| CLONE OF slc7A8 (SOLUTE CARRIER FAMILY 7) GENE AND ITS EXPRESSION ANALYSIS IN CARASSIUS AURATUS TISSUES |
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XIONG Gang1,2, LIU Zhen1, WANG Xiao-Qing3, LU Shuang-Qing1, ZHANG Jian-She1, XIE Di-Zhi1,3, LUO Xiao-Hua1,3
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1.Department of Biotechnology＆Environment Science, Changsha University;2.unan Biological and Electormechinacial Polytechinc;3.College of Animal Science and Technology, Hunan Agricultural University
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| Abstract: |
| We studied the information of Carassius auratus slc7A8 and analyzed the regulation of slc7A8 mRNA expression in different tissues. The full cDNA sequence of slc7A8 of C. auratus was cloned by using RT-PCR and RACE-PCR. It had 2709 nucleotides, including 5′UTR of nucletides and 3′ URT of nucleotides. Its open reading frame had 1593 nucleotides, encoding a 530-amino-acid peptide. The sequence of C. auratus slc7A8 had the highest similarity with Danio rerio (89.9%). The homology with other animals was between 70.1% and 74.8%. Interestingly, the amino acid sequence of C. auratus slc7A8 had the highest similarity with D. rerio (95.5%), and the similarity with others varied from 80.6% to 82.1%. This protein had 12 helix transmenbrane regions. The homology of transmenbrane region AA had the highest similarity among all animals. The abundances of slc7A8 mRNA were measured in foreintestine and midintestine, brain, hind intestine, muscle, kidney, gill, heart, and liver, from high to low.
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| Key words: Carassius auratus, slc7A8 cDNA, Molecular characteristics, slc7A8 mRNA expression abundance |
