剑白香猪葡萄糖转运蛋白4 cDNA的克隆及序列分析

为了在基因水平上给动物的营养代谢研究提供基础性资料,为系统研究剑白香猪这一优质猪种提供参考,以剑白香猪肌肉组织中的总RNA为模板,克隆了剑白香猪的GLUT-4cDNA序列,并对其序列进行了分析。结果表明:克隆片段总长为1 616bp,包含一个长1 530bp的开放阅读框(ORF),编码509个氨基酸残基的蛋白;该蛋白的相对分子质量为54.809 4kDa,等电点为6.98,包含30个功能位点,即3个N-糖基化位点、1个cAMP和cGMP依赖性蛋白激酶磷酸化位点、4个酪蛋白激酶Ⅱ磷酸化位点、2个酪氨酸激酶磷酸化位点、4个蛋白激酶C磷酸化位点、2个酰胺化位点、12个N-豆蔻酰化位点、1个糖转运蛋白signature 1位点和1个糖转运蛋白signature 2位点;剑白香猪与牛、兔、小家鼠、人、马、黑猩猩、褐家鼠、非洲爪蟾GLUT-4基因编码序列的同源性分别为91.76%、90.52%、87.91%、90.52%、92.29%、90.46%、87.52%和65.82%,氨基酸序列的同源性分别为94.7%、95.68%、94.11%、95.09%、94.89%、95.28%、94.5%和68.31%。

cDNA Cloning and Sequence Analysis of GLUT-4 from the Jianbai Xiang-pig

With the purpose of providing the theoretical basis to nutrimental metabolism at the level of gene and offering reference to systematically study Jiangbai Xiang-pig,cDNA of GLUT-4 was amplified by RT-PCR from total RNA of Jiangbai Xiang-pig musculature cell,then the RT-PCR products were cloned and sequenced.The results showed that the length of the fragment was 1 616 bp,containing an ORF of 1 530 bp,which encoded a protein of 509 amino acid residues.The pI of the protein was 6.98 and its molecular weight was 54.8094 kDa.Protein prediction shows that the protein contain 30 function sites: three N-glycosylation sites,one cAMP-and cGMP-dependent protein kinase phosphorylation site,four casein kinase II phosphorylation sites,two tyrosine kinase phosphorylation sites,four protein kinase C phosphorylation sites,two Amidation sites,12 N-myristoylation sites,one sugar transporter proteins signature1,noe sugar transporter proteins signature 2.The coding sequence and the amino acid sequence of GLUT-4 gene from Jiangbai Xiang-pig were compared with those of cattle,rabbit,mouse,human,horse,chimpanzee,norway rat,African clawed frog respectively.The homologies of the coding sequence were 91.76 %,90.52 %,87.91%,90.52%,92.29 %,90.46%,87.52% and 65.82% respectively.The homologies of the amino acid sequence were 94.7%,95.68%,94.11%,95.09%,94.89%,95.28%,94.5% and 68.31% respectively.