花生Clp蛋白酶基因(AhClpP)的克隆与序列分析  

Cloning and Analyzing of Caseinolytic Protease Gene from Arachis hypogaea L.

作　　者：纪鸿飞[1] 彭振英[2] 马敬[1] 毕玉平[1,2,3] 

机构地区：[1]山东师范大学生命科学学院,山东济南250014 [2]山东省农业科学院高新技术研究中心,山东省作物与畜禽品种改良生物技术重点实验室,山东济南250100 [3]农业部黄淮海作物遗传改良与生物技术重点实验室,山东济南250100

出　　处：《华北农学报》2010年第B12期5-8,共4页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金(30871541)

摘　　要：Clp蛋白酶通过蛋白质水解作用来清除细胞内由逆境引起的异常和具有潜在毒性的蛋白或多肽,从而保证细胞正常的生理功能。从优质大花生鲁花14种子不同发育时期混合cDNA文库中发现一条序列,全长为1 212 bp,3′端含有polyA尾,通过blastx搜索得知其为Clp蛋白酶基因。该序列与GenBank中EZ736390.1的序列相似性达到99%。以该花生cDNA序列为模板设计引物,以花生幼嫩种子cDNA为模板克隆得到花生Clp蛋白酶基因。序列分析表明,AhClpP基因的开放阅读框长度为843 bp,编码280个氨基酸,预测其分子量为30.9 kDa,等电点为9.07,编码的蛋白包含S14-ClpP-2保守结构域,无信号肽,定位于线粒体。通过与其他植物Clp蛋白酶的氨基酸序列比对,发现与拟南芥、蓖麻的Clp蛋白酶同源性较高。花生AhClpP基因的克隆为进一步研究其生物学功能和应用奠定了基础。Clp protease controls ordinary metabolic processes by removing abnormal and toxic proteins or peptides caused by stress.By constructing the cDNA library of different stages seeds of peanut cultivar Luhua 14,we selected a sequence of 1 212 bp with polyA in 3′end.By blastx we found that it was a ClpP gene and had 99% similarity with EZ736390.1 in the Genbank.Based on tender peanut seed cDNA,we designed primers and cloned AhClpP gene.Sequence analysis demonstrates that the cDNA of AhClpP gene has a single open reading frame of 843 bp encoding 280 amino acids.The predicted protein has an molecular mass of 30.9 kDa and an isoelectric point of 9.07.The estimated protein contains a S14_ClpP_2 conserved domain,has no signal peptide and sublocates in mitochontria.Compared with other ClpP members,AhClpP has high homology with its counterpart of Arabidopsis thaliana and Ricinus communis.The cloning of AhClpP gene provides foundation for further study of its biological function and application.

关 键 词：花生 Clp蛋白酶 克隆 序列分析 

分 类 号：Q78[生物学—分子生物学]