• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

龙眼MYB14的生物信息学分析及其基因表达载体构建

Bioinformatics and Construction of Expression Vector for Longan MYB14

  • 摘要:
      目的  克隆龙眼MYB-related基因家族中MYB14基因,并对其进行生物信息学分析和基因表达载体的构建,为进一步研究该基因的生物学功能提供基础数据。
      方法  通过转录组测序及生物信息学分析得到龙眼MYB14基因全长序列,进一步设计特异引物对其开放阅读框(ORF)全长序列进行克隆和生物信息学分析,同时构建超表达载体。
      结果  MYB14基因属于MYB-related家族的TBP-like亚家族,ORF长度831 bp,编码276个氨基酸,有一个MYB结合位点。该蛋白属于非跨膜亲水蛋白,亚细胞定位预测定位于细胞质,存在57个氨基酸磷酸化位点。其二级结构主要由无规则卷曲以及α-螺旋构成,二级结构与三级结构预测结果高度一致,同源基因进化树分析表明该基因与大麻亲缘关系最近,同时成功构建了植物超表达载体pBI121-MYB14。
      结论  本研究克隆得到了龙眼MYB14基因,成功构建表达载体pBI121-MYB14,为该基因功能的研究奠定了坚实的基础。

     

    Abstract:
      Objective  MYB14 in Longan was cloned, its bioinformatics studied, and expression vector constructed.
      Method  Full-length sequence of MYB14 was obtained by transcriptome sequencing and bioinformatics analysis. Accordingly, specific primers for the cloning were designed, bioinformatics on the open reading frame sequence established, and overexpression vector constructed.
      Result  In the TBP-like subfamily of MYB-related family of genes, MYB14 had an 831 bp ORF encoding 276 amino acids with one MYB binding site. It was a non-transmembrane hydrophilic protein with 57 amino acid phosphorylation sites in the cytoplasm. Its secondary structure was irregular crimp and α-helix. The predicted secondary and tertiary structures were highly consistent. The phylogenetic tree on the gene of longan showed a homology with that of cannabis.
      Conclusion   MYB14 of Longan was successfully cloned with the constructed expression vector, PBI121-MYB14, which provided reference for the further study on the function of MYB14.

     

/

返回文章
返回