苹果属山荆子MbNramp1基因克隆、序列与表达分析  

MbNramp1 Gene Cloning,Sequence Analysis and Expression Analysis in Malus baccata (L.) Borkh.

作　　者：肖海华[1,2,3] 印莉萍[3] 韩振海[1,2] 

机构地区：[1]中国农业大学园艺植物研究所,北京市果树逆境生理与分子生物学重点实验室,北京100193 [2]四川农业大学资源环境学院,四川雅安625014 [3]首都师范大学生命科学学院,北京100048

出　　处：《园艺学报》2010年第9期1409-1415,共7页Acta Horticulturae Sinica

基　　金：国家转基因专项(2009ZX08009-122B);国家自然科学基金项目(30971982;30800741;30700545;30671441);中国博士后科学基金特别资助项目(200902153)

摘　　要：以苹果属山荆子为试材,根据Nramp1同源序列保守区设计两对简并引物进行PCR,结合RACE技术获得了3'末端和5'末端片段,将3段片段序列拼接,根据拼接序列获取了MbNramp1基因全长cDNA序列。MbNramp1cDNA长2090bp,包含一个长度为1656bp的开放阅读框,编码551个氨基酸的多肽,分子量约为59.7kD。该基因编码的蛋白是一个膜蛋白,76%以上的氨基酸为非极性。MbNRAMP1与二价铁、锰转运蛋白NRAMP基因家族保守性很高,具有NRAMP1金属转运蛋白家族基因典型特征,即含有推断的N–端相连的糖基化位点、10个预测的跨膜结构域(TMs),在第6和第7跨膜结构域间有一个相同的转运基序(CTM)。低铁胁迫时MbNramp1在根中表达量增加。Degenerate primers corresponding to the conserved motifs of NRAMP1 in plants were used to amplify specific DNA fragments from Malus baccata（L.）Borkh.roots cDNA.Then the gene specific primers were designed based on the obtained specific DNA fragments,the 3＇ end and 5＇ end fragments were amplified by RACE.A 2 090 bp full length cDNA MbNramp1 containing a 1 656 bp ORF was obtained based on specific DNA fragments.MbNramp1 encodes a polypeptide of 551 amino acids with a predicted molecular mass of 59.7 kD and a membrane protein with more than 76% non-polar amino acids.All conserved features of NRAMP described previously were present in the predicted MbNRAMP1 sequence,such as putative N-linked glycosylation sites,10 transmembrane domains（TMS）and a consensus transporter motif（CTM）located between Ⅵ and Ⅶ TM.Expression of MbNramp1 was increased in roots under iron deficiency,and its mRNA accumulation patterns differed with the induction time.

关 键 词：山荆子 Nramp1 MbNramp1 铁转运蛋白 锰转运蛋白 

分 类 号：S661[农业科学—果树学]