木薯磷酸烯醇式丙酮酸羧化酶基因全长cDNA克隆及表达分析

Cloning and Expression Analysis of Phosphoenolpyruvate Carboxylase Gene pepc from Cassava (Manihot esculenta Crantz)

摘要:

应用巢式PCR 从木薯栽培种(Manihot esculenta) Arg7 和野生种W14 (M. esculenta subsp. flabellifolia)叶片中克隆到磷酸烯醇式丙酮酸羧化酶基因pepc 全长cDNA,GenBank 序列号为JN387052 和JN387053。cDNA 全长2945 bp,含1 个2895 bp 的开放阅读框,预测编码的蛋白含964 个氨基酸,具有磷酸烯醇式丙酮酸羧化酶保守结构域。木薯PEPC 与麻疯树和蓖麻的PEPC氨基酸序列具有高度同源性。表达分析表明pepc 在W14 和Arg7 叶中的表达量最高,其次是须根和块根,茎中最低。单日表达量动态分析表明,Arg7 叶中pepc 总体表达量高于W14,但是16∶00 后W14 高于Arg7,推测两种木薯pepc 调控区域存在差异。

Abstract:

The full-length cDNAs of phosphoenolpyruvate carboxylase gene pepc were cloned from cassava (Manihot esculenta) cultivar Arg7 and wild species (M. esculenta subsp. flabellifolia) W14 by using Nest-PCR method, respectively, with corresponding GenBank accession number of JN387053 and JN387052. Both of two cDNA sequences were 2945 bp in length including a ORF of 2895 bp, predicting to encode a protein with 964 amino acids and containing typical conserved sequences/domains of pepc gene. Cassava PEPC had high homologous with that in Ricinus communis and Jatropha curcas. The pepc expression was the highest in leaves of W14 and Arg7, followed fibrous roots and tuberous roots, and the lowest expression was in stems. The oneday dynamic expression analysis showed that the pepc expression in Arg7 leaves was higher than in W14 before 16∶00, while it was higher in W14 than Arg7 after 16∶00. Therefore, it suggested that the regulatory regions of pepc gene in two cassava species were different.