中国春硬度基因的克隆与序列分析  

Cloning and Sequence Analysis of Hardness Genes in Chinese Spring

作　　者：罗立廷[1] 陶红梅[1] 杨广笑[1] 何光源[1] 郑重 

机构地区：[1]华中科技大学中英HUST-RRes基因工程和基因组学联合实验室,湖北武汉430074 [2]湖北省洪湖市第一中学,湖北洪湖437200

出　　处：《生物技术》2007年第6期1-4,共4页Biotechnology

基　　金：国家863课题资助(编号:2002AA224031)

摘　　要：目的:克隆、分析puroindoline a(Pina)、puroindoline b(Pinb)和grain softness protein(gsp)这三种控制小麦籽粒硬度的主要基因。方法:根据已报导的小麦3种基因的保守序列,设计合成了3对特异性引物,对六倍体软质小麦中国春基因组DNA进行基因扩增、克隆和序列测定,得到了3个基因的全长。结果:其ORF分别是447bp、447bp和495bp,编码的蛋白质全长分别为148aa、148aa和164aa。3种蛋白质都含有谷类作物所特有的19aa的信号肽,10个半胱氨酸所形成的5个二硫键结构,PINA和PINB蛋白有麦类作物所特有的富含色氨酸的结构域。结论:与粗山羊草的pina、pinb和gsp相比较,其核苷酸同源性为99.8%、95.8%和99.4%,氨基酸的同源性高达99.3%、96.6%和98.8%。3种与硬度相关的基因的分离与克隆丰富了小麦种质遗传资源库,为弄清影响小麦籽粒硬度的机制及其抗菌活性奠定了基础。Objective： Clone and sequence puroindoline a （Pina）, puroindoline b （ Pinb ） and grain sofiness protein （gsp） the three major genes controlling grain hardness in wheat. Methods： According to the conserved regions of three genes reported, three pairs of specific primers were designed and used to amplify them from genomic DNA of soft hexaploid Chinese spring＇ s young leaves, Results： Through gene cloning and sequencing, they were 447bp, 447bp and 495bp in size, encoding 148, 148 and 164 amino acid residues respectively. Three proteins had a signal peptide, which were specific in cereal crops, five disulphide bonds formed by ten cysteines and both pina and pinb contained the tryptophan - rich domain, which were specific in wheat. Conclusion： Compared with pina, pinb, gsp from Aegilops tauschii, they shared 99.8 %, 95.8 % and 99. 4 % homologies in nucleotide sequence and 99.3 %, 96.6% and 98.8 % homologies in amino acid sequence respectively. The isolatiom of three major genes relating to grian hardness enriched the genetic resources for cultivated wheat and laid a fundamental basis for understanding the reason influencing the wheat hardness.

关 键 词：中国春 PUROINDOLINE GSP 籽粒硬度 克隆 

分 类 号：Q785[生物学—分子生物学]