Cloning and Identification of A New Na^+/H^+ Antiporter Gene ZmSOS1 in Maize(Zea mays L.)  

玉米Na^+/H^+逆向转运蛋白基因ZmSOS1的克隆与鉴定(摘要)(英文)

作　　者：赵祥强[1] 

机构地区：[1]南通大学生命科学学院,江苏南通226007

出　　处：《Agricultural Science & Technology》2009年第6期57-62,158,共7页农业科学与技术（英文版）

基　　金：Supported by the Natural Science Foundation of the Department of Educationof Jiangsu Province(07KJD180168);the Doctoral ScienceStarting Foundation of Nantong University;And the Openning Subjectof Plant Functional Genomics Key Laboratory of Jiangsu Province~~

摘　　要：[ Objective] The study aimed to clone and identify Na^＋/H^＋ antiporter genes in maize, and provided the information for characterizing the function of such genes in abiotic stress tolerance of maize. Method The in silico cloning, RT-PCR, and bioinformatics analysis were used in this study. Result By in sifico cloning, a plasma membrane Na^＋/H^＋ antiporter gene, named as ZmSOS1 （EMBL accession No. BN001309）, was cloned from maize （ Zea mays L. ）. ZmSOS1 has an open reading frame （ORF） of 3 411 bp which encoded a protein of 1 136 amino acids. By multiple sequence alignment analysis, it showed the predicated peptide of ZmSOS1 were 61% and 82% identities in amino acids to the plasma membrane Na^＋/H^＋ antiporter AtSOS1 and OsSOS1, respectively. The RT-PCR analysis revealed that ZmSOS1 could be significantly up-regulated by salt stress, which indicated ZmSOS1 might play a role in salt tolerance of maize. Conclusion ZmSOS1 is a putative plasma membrane Na^＋/H^＋ antiporter gene and may play a role in abiotic stress tolerance of maize.[目的]研究玉米Na^+/H^+逆向转运蛋白基因ZmSOS1的克隆与鉴定,为研究该基因在玉米逆境信号转导与生长发育中的作用提供参考。[方法]采用电子克隆、RT-PCR、生物信息学方法,从玉米基因组中克隆1个质膜型Na^+/H^+逆向转运蛋白基因,并鉴定该基因编码产物的跨膜结构预测以及在盐胁迫下的表达模式等信息。电子克隆具体步骤:将水稻质膜型Na^+/H^+逆向转运蛋白OsSOS1的氨基酸序列作为种子序列输入GenBank,对玉米基因组数据库和EST数据库进行tBLASTN分析,结果检索到含有候选基因的基因组序列AC186524和1批高度相似的玉米EST。为确定候选基因的编码区,将上述EST序列进行拼接,并将拼接所得的contig通过BLASTN分析定位到基因组序列AC186524中。为获得完整的编码区序列,利用与contig间隔区相对应的水稻OsSOS1蛋白的氨基酸序列,进一步对玉米基因组序列进行tBLASTN分析;同时,结合GENSCAN软件的基因预测结果,最终确定了候选基因的编码区序列。最后,通过比较编码区序列和基因组序列,确定ZmSOS1基因的外显子和内含子结构。[结果]利用电子克隆方法,从玉米中克隆出1个质膜型Na^+/H^+逆向转运蛋白基因ZmSOS1(编码序列反向互补于AC186524中的78 964～96 731 bp处);ZmSOS1基因的开放阅读框长3 411 bp,编码1 136个氨基酸的蛋白。ZmSOS1蛋白的理论等电点pI=6.46,分子质量为126.2kDa,理论半衰期大于10 h,不稳定参数为41.74,属于不稳定蛋白。通过在线软件TMHMM2.0和TMPRED分析,发现ZmSOS1蛋白含有12个跨膜结构区,且有一个长的高度亲水性的羧基端尾巴,暗示质膜型Na^+/H^+逆向转运蛋白在进化上是较为保守的。ZmSOS1基因至少含有21个内含子,如果将ZmSOS1基因的结构分别与水稻、拟南芥和苔藓植物(Phycomitrella patens)的同源基因OsSOS1、AtSOS1、PpSOS1相比较,可以发现质膜型Na^+/H^+逆向转运蛋白基因编码区的结构是比较保守

关 键 词：Zea mays Na^＋/H^＋ antiporter In silico cloning Bioinformatics analysis 

分 类 号：S513[农业科学—作物学]