小麦Clp蛋白酶基因(TaClpP)的克隆与序列分析  

Cloning and Sequence Analysis of Clp Protease Gene(TaClpP) from Wheat(Triticum aestivum L. )

作　　者：张斌[1] 杨连群[1] 边斐[1] 李娜娜 范仲学[1] 宫永超 蒲艳艳 丁汉凤 彭振英[1] 

机构地区：[1]山东省农业科学院生物技术研究中心/山东省作物遗传改良与生态生理重点实验室,山东济南250100 [2]山东省农作物种质资源中心,山东济南250100

出　　处：《山东农业科学》2014年第6期1-5,共5页Shandong Agricultural Sciences

基　　金：国家重大科技专项转基因新品种培育"解析双特异性蛋白激酶介导的信号传递网络在提高植物抗性中的分子作用机制"(2014ZX0800916B-003);"十二五"农村领域国家科技计划课题"小麦种质资源发掘与创新利用"(2013BAD01B02-13);山东省科技发展计划"山东省近六十年主推小麦品种磷效率演变及磷高效品种的筛选与利用"(2012G0021031)

摘　　要：Clp蛋白酶(ClpP)普遍存在于各种生物体内,在蛋白质代谢调控过程中发挥着极为重要的作用。本研究利用同源克隆和RACE方法从小麦叶片中获得了TaClpP基因(GenBank No.KJ541961)的全长cDNA序列。序列分析表明,该基因含有897 bp的完整阅读框,编码蛋白含有298个氨基酸,预测相对分子量为32.6 kD,等电点为9.79。该蛋白含有一个保守的S14-ClpP-2结构域,无信号肽,定位于叶绿体。与其他植物Clp蛋白酶的氨基酸序列比对发现,TaClpP与节节麦、乌拉尔图小麦的Clp蛋白酶相似性较高。另外,以小麦基因组DNA为模板,通过PCR扩增获得了TaClpP的基因组序列,长度为3 256 bp,包含9个外显子、8个内含子。Clp protease （ ClpP） widely exists in various prokaryotes and eukaryotes , and plays important roles in the regulation of protein metabolism .In this study , a TaClpP gene （ GenBank No .KJ541961 ） was cloned from wheat （ Triticum aestivum L.） leaves by homology cloning strategy and RACE method .The cDNA sequence analysis showed that TaClpP had a single open reading frame of 897 bp; the predicted protein had 298 amino acids and the predicted molecular mass was 32 .6 kD;the isoelectric point was 9 .79 .This predic-ted protein contained a conserved S14-ClpP-2 domain motif, and had no signal peptide in its N -terminus. The supposed subcellular location of TaClpP was in chloroplast .Compared with other plants ’ ClpP members, TaClpP had high homology with its counterpart of Aegilops tauschii and Triticum urartu.In addition, the TaClpP genome sequence was obtained by PCR amplification , which contained 9 exons and 8 introns, and the length was 3 256 bp.

关 键 词：小麦 Clp蛋白酶基因 基因克隆 序列分析 

分 类 号：Q785[生物学—分子生物学]