红地球葡萄VvWRKY11VvWRKY75基因克隆及序列分析

Cloning and bioinformatics analysis of VvWRKY11 and VvWRKY75 from Red Earth Grape

  • 摘要:
    目的 克隆红地球葡萄果实中转录因子VvWRKY11VvWRKY75全长,并研究其序列特征,为研究该转录因子在葡萄果实中的抗病作用和功能奠定基础。
    方法 依据课题组前期的采前水杨酸(SA)喷施葡萄果实转录组测序数据,筛选并通过同源克隆得到受SA剧烈诱导果实应答灰葡萄孢菌显著差异表达的2个WRKY转录因子VvWRKY11VvWRKY75的全长cDNA序列,进一步运用生物信息学方法对VvWRKY11VvWRKY75进行序列分析。
    结果 转录因子VvWRKY11VvWRKY75的cDNA全长分别为1 075和965 bp,开放阅读框分别为894和570 bp,分别编码297和189个氨基酸,终止密码子分别为TGA和TAA,预测相对分子量分别为32.36和21.26 kD,理论等电点分别为9.73和9.13,均属于不稳定亲水性蛋白。葡萄VvWRKY11和VvWRKY75蛋白二级结构主要由无规则卷曲和α-螺旋组成,两者基因启动子含有W-box、MeJA、SA和ABA等响应元件。
    结论 获得红地球葡萄转录因子VvWRKY11VvWRKY75基因全长编码序列,探明该序列的结构特征。

     

    Abstract:
    Objective To clone the full-length transcription factors VvWRKY11 and VvWRKY75 in Red Earth Grape berries and study their sequence characteristics in the hope of laying the foundation for the study of the role and function of these transcription factors in grape berries' disease resistance.
    Methods Based on the transcriptome sequencing data of pre-harvest SA sprayed grapes, we screened and cloned two WRKY transcription factors, VvWRKY11 and VvWRKY75, which were significantly differentially expressed in SA-induced fruit in response to Staphylococcus griseus, and then further analyzed the sequences of VvWRKY11 and VvWRKY75 by bioinformatics methods.
    Results The full length of VvWRKY11 and VvWRKY75 gene sequences were 1 075 and 965 bp, respectively, with open reading frames of 894 and 570 bp, encoding 297 and 189 amino acids, respectively, and with termination codons TGA and TAA, respectively, and with predicted relative molecular weights of 32.36 and 21.26 kD, respectively, and theoretical isoelectric points of 9.73 and 9.13, both of which were unstable hydrophilic proteins. The secondary structures of both grape VvWRKY11 and VvWRKY75 proteins consist mainly of irregularly coiled and α-helices, and the promoters of both genes contain hormone-responsive elements such as the W-box, methyl jasmonate, salicylic acid and abscisic acid.
    Conclusion The full-length coding sequences of the Red Earth Grape transcription factors VvWRKY11 and VvWRKY75 genes are obtained, and the structural features of the sequences are explored.

     

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