大豆中GmKAB1基因的克隆和信息学分析  

Molecular cloning and bioinformatics analysis of GmKAB1 from soybean[Glycine max( L. ) Merri]

作　　者：朱晓玲[1,2] 郝青南[1,2] 陈海峰[1] 王程[1,3] 陈李淼[1] 沙爱华[1] 陈水莲[1] 周蓉[1] 周新安[1] 

机构地区：[1]中国农业科学院油料作物研究所/农业部油料作物生物学重点开放实验室,湖北武汉430062 [2]中国农业科学院研究生院,北京100081 [3]南京农业大学/国家大豆改良中心/国家作物遗传与种质创新重点实验室,江苏南京210095

出　　处：《中国油料作物学报》2014年第1期18-24,共7页Chinese Journal of Oil Crop Sciences

基　　金：国家转基因生物新品种培育重大专项(2013ZX08004-005);国家自然科学基金项目(30871554,30900906);中国农业科学院公益性院所科研基金(1610172011006)

摘　　要：为探讨GmKAB1在大豆处于低钾胁迫下的表达水平,以大豆钾高效品系油06-71和钾敏感型品系衡春04-11为试验材料,设置低钾胁迫试验,分别在处理后0.5h、2h、6h、12h、3d、6d、9d和12d取样提取RNA,利用Real time-PCR检测各时间段GmKAB1基因的表达量。结果显示GmKAB1基因在地下部分表达比地上部持续时间更长,地下部相对表达倍数最高为3.5,较地上部相对表达倍数更高。克隆目的基因并对基因序列进行同源性及生物信息学分析,结果表明,与GmKAB1基因相似性在30%以上的同源基因有45个,GmKAB1在进化树中的位置与Glyma20g19000最近;GmKAB1编码蛋白为稳定的可溶性蛋白,具有2个保守结构域,多个磷酸化位点,表明在受到低钾胁迫后该基因编码的β亚基与α亚基结合调控电压门控钾离子通道,对大豆从根部获取钾离子可能起着关键作用。： A low - K - tolerant soybean line （ You06 - 71 ） and a low - K - sensitive line （ Hengchun04 - 11 ） were selected as plant materials to study the expression level of GmKAB1 in low potassium stress condition. RNA was extracted at 0.5h, 2h, 6h, 12h, 3d, 6d, 9d and 12d after stress treatment, and Real -time PCR was carried out respectively. The results showed that the expression of GmKAB1 in roots were lasted longer than that in shoots, and the relative expression levels in roots were significantly higher than that in shoots, and the highest level in roots was up to 3.5 times. Homology and bioinformatics analysis after cloning the target sequencing from the two lines in- dicated that comparing with GrrtKAB1, there were 45 homologous genes with more than 30% similarity, and Gly- ma20gl9000 was closest to GmKAB1 in the phylogenetic tree. The protein encoded by GmKAB1 was stable and soluble, which contained two conversed structural domains, possessed multiple phosphorylation sites. These indicated that GmKAB1 polypeptide formed a close physical association with 15 subunit and ct subunit to control voltage - ga- ted K ＋ channel. In conclusion, GmKAB1 might have played a pivotal role in potassium absorption in soybean and had important value in research.

关 键 词：大豆 钾离子通道基因 克隆 实时荧光定量PCR 生物信息学分析 同源性分析 系统发育分析 

分 类 号：S565.103[农业科学—作物学]