东方山羊豆肌醇-1-磷酸合酶基因的克隆与分析  

Cloning and Analysis of Myo-Inositol-1-Phosphate Synthases in Galega Orientalis

作　　者：李春艳[1] 宋清晓[2] 刘建宁[1] 高洪文[2] 

机构地区：[1]山西省农业科学院畜牧兽医研究所,山西太原030032 [2]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《家畜生态学报》2010年第5期17-22,共6页Journal of Domestic Animal Ecology

基　　金：国家科技支撑计划(2007BAD56B02);山西省农科院畜牧兽医研究所基金项目(YGG0853)

摘　　要：探索新的基因和基因组合在植物耐盐性的研究中是至关重要的。本试验根据在不同植物中功能相同的同源基因保守序列设计引物,以250 mM NaCl胁迫后东方山羊豆(Galega.orientalis L.)幼嫩叶片中所提取的总RNA为模板,采用RT-PCR及RACE方法从东方山羊豆中克隆了一种新的肌醇-1-磷酸合成酶基因GoMIPS,cDNA全长1 858 bp,3'非编码区为256 bp,5'非编码区为69 bp,开放阅读框为1 533 bp,编码510个氨基酸。与已报道物种的MIPS基因氨基酸序列比较分析表明,GoMIPS与其他植物MIPS基因氨基酸相似性高达87%～95%,GoMIPS推导的氨基酸序列中含有植物MIPS基因所具有典型的NAD+结合区(GWGGNNG)和底物结合位点(SYNHLGNNDG),且9种植物NAD+结合区位置比较保守,均起始于第68个氨基酸。这一结果暗示功能结构域在进化过程中具有较高的稳定性。Exploring novel genes and gene combinations are of primary importance in plant salt-tolerance.The primers were designed according to the conserved region of the homologous gene in other plants.GoMIPS was amplified by RT-PCR and RACE from the total RNA from young leaf of Galega orientalis with 250 mmol/L sodium chloride.Sequence analysis revealed that the full-length of GoMIPS gene was 1858 bp,the open reading frame of GoMIPS was 1533bp and encoded a putative protein of 510 amino acids residues,and including 256 bp 3＇untranslated region,69 bp 5＇untranslated region.When deduced amino acid sequences were compared with homologous sequences from other organisms,GoMIPS was found sharing sequence similarity（87%-95%） with plant MIPS genes.In addition,GoMIPS also contained a typical NAD＋-binding domain＂GWGGNNG＂ and substrate binding site＂SYNHLGNNDG＂,which were identical to those of other MIPS from plants.Further analysis showed that the location of NAD＋-binding domain was relatively conserved,which all originated the first 68 amino acids.The results suggested that functional domains in the evolution of high stability.

关 键 词：东方山羊豆 肌醇-1-磷酸合成酶 CDNA末端快速扩增 基因克隆 

分 类 号：S811.5[农业科学—畜牧学]