甜荞木瓜类半胱氨酸蛋白酶基因FeRD21的克隆与表达分析（英文）  

Clone and Expression Analysis of a Papain-like Cysteine Protease Gene(FeRD21)in Fagopyrum esculentum

作　　者：方正武[1] 李来运[2] 李晓方[1] 刘志雄[1,2] 

机构地区：[1]长江大学作物遗传育种研究所暨农作物多基因型种群育种技术中心,湖北荆州434025 [2]长江大学园艺园林学院,湖北荆州434025

出　　处：《西北植物学报》2015年第3期459-464,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(31101202);国家公益性行业(农业)科研专项(201303008);湖北省重点(优势)学科作物学(长江大学)

摘　　要：该研究以旱区小杂粮作物甜荞(Fagopyrum esculentum)为材料,采用同源克隆、RACE技术和实时荧光定量RT-PCR方法,对其半胱氨酸蛋白酶基因(FeRD21)进行了分离和表达分析。结果表明:(1)FeRD21基因cDNA全长1 750bp,包含1个1 407bp的完整开放阅读框,编码468个氨基酸。(2)蛋白序列比对发现,甜荞FeRD21全酶包括信号肽、N末端自主抑制前体区域、蛋白酶、脯氨酸富含结构域和C末端颗粒体蛋白结构域,同时,其蛋白酶结构域包含1个木瓜类蛋白酶家族保守的催化三连体活性位点:Cys168-His304-Asn324。(3)分子系统发生分析证实,其与拟南芥的RD21一致性最高,属类RD21半胱氨酸蛋白酶类。(4)基因表达分析表明,FeRD21能被干旱、高盐、ABA和衰老胁迫诱导。Fagopyrum esculentum（buckwheat,Polygonaceae）is a multi-food-use pseudocereal with healing benefits and is growing on arid areas.（1）Based on homology and RACE method,aRD21 orthologous gene from buckwheat was isolated and identified.The RD21 homologous gene fromF.esculentumtranscript was1 750 bp and contained a 1 407 bp ORF（Open Reading Frame,ORF）encoding 468 amino acids.（2）Protein sequence alignment and phylogenetic analyses grouped FeRD21 into PLCPs subfamily members which carry a C-terminal granulin domain.（3）The protease of FeRD21 was highly conserved and harbored the conservation sites of catalytic residues Cys168-His304-Asn324.（4）Expression analysis suggested that FeRD21 was up-regulated by salt,dehydration,ABA,and senescent treatments,which showed a different way in response to stresses with RD21 in Arabidopsis.Our results indicated that FeRD21 might be involved the stress-responsive pathways in F.esculentum.

关 键 词：非生物胁迫 甜荞 木瓜类半胱氨酸蛋白酶 FeRD21 

分 类 号：Q786[生物学—分子生物学]