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。
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.
出处
《海洋与湖沼》
CAS
CSCD
北大核心
2010年第6期935-940,共6页
Oceanologia Et Limnologia Sinica
基金
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
