园艺学报 ›› 2008, Vol. 35 ›› Issue (7): 1017-1022.

• 蔬菜 • 上一篇    下一篇


徐慧妮1 ;王秀峰1,2*;孙旭东1;杨凤娟1,2;杜栋良1   

  1. 1山东农业大学园艺科学与工程学院; 2作物生物学国家重点实验室,山东泰安 271018)
  • 收稿日期:2007-12-12 修回日期:2008-05-15 出版日期:2008-07-25 发布日期:2008-07-25
  • 通讯作者: 王秀峰

Bioinformatics Analysis and Prokaryotic Expression of MAPK in Cucumber

XU Hui-ni1, WANG Xiu-feng1,2*, SUN Xu-dong1, YANG Feng-juan1,2, and DU Dong-liang1   

  1. (1College of Horticulture Science and Engineering , Shandong Agricultural University; 2State Key Laboratory of Crop Biology, Tai 'an, Shandong 271018, China )
  • Received:2007-12-12 Revised:2008-05-15 Online:2008-07-25 Published:2008-07-25
  • Contact: WANG Xiu-feng


利用RT-PCR技术结合RACE技术,从NO3-胁迫下的黄瓜根中克隆出促分裂原活化蛋白激酶(mitogen-activated protein kinase, MAPK)基因的同源序列,命名为CsNMAPK,GenBank注册号为DQ812086。生物信息学分析表明,该基因全长1 636 bp,开放阅读框(ORF)1 113 bp,编码370个氨基酸。亚细胞定位预测其编码蛋白可能定位于细胞质;跨膜结构分析表明该基因有一个强的跨膜螺旋结构;PlantCare分析结果显示该基因序列具有脱落酸诱导、茉莉酸诱导、赤霉素诱导、水杨酸诱导、伤害诱导等顺式作用元件。成功构建原核表达载体,并转化E.coli BL21(DE3),SDS-PAGE检测结果显示表达了一个约46 kD的蛋白。为进一步揭示黄瓜MAPK基因在NO3-胁迫中的作用奠定基础。

关键词: 黄瓜, 促分裂原活化蛋白激酶, NO3-胁迫, 生物信息学分析, 原核表达


A cucumber cDNA designated CsNMAPK (GenBank accession NO. DQ812086) was isolated by RT-PCR and RACE. Bioinformatics analysis indicated that the full-length cDNA sequence was 1 636 bp, which contained an open reading frame (ORF) of 1 113 bp and encoded a protein of 370 amino acid residues. The subcellular localization analysis predicted that the protein was in the cytoplasm and there was one strong inside to outside transmembrane helix. PlantCare analysis indicated that there were abscisic acid, MeJA, salicylic acid cis-acting responsive elements and gibberellin, wound-responsive elements in CsNMAPK. The CsNMAPK was cloned into pET-30a(+) vector to construct recombination prokaryotic expression vector pET-CsNMAPK. After transformed to E.coli BL21 and induced by isopropyl-β-D-thiogalactopyranoside (IPTG), recombinant protein about 46 kD was expressed in pET-CsNMAPK system and separated by SDS-PAGE electrophoresis. The research will be helpful to study the function of cucumber MAPK gene in response to NO3- stress.

Key words: cucumber, MAPK, NO3-stress, bioinformatics analysis, prokaryotic expression