辣椒CaCBF1A基因的克隆及非生物胁迫下表达分析

Cloning and Expression Analysis of CaCBF1A Gene from Capsicum annuum L. under Abiotic Stress

为揭示Ca CBF1A基因在辣椒抗逆机制中发挥的功能,对辣椒Ca CBF1A基因进行克隆与分析。以豫椒101为材料,根据参考基因组序列设计引物,通过PCR技术从辣椒基因组c DNA中获得CBF基因Ca CBF1A。经生物信息学分析,该基因具有一个完整的ORF(471 bp),编码156个氨基酸。Ca CBF1A编码的蛋白质包含保守的AP2 DNA结合域。对其亚细胞定位、跨膜结构进行分析,预测其定位在叶绿体中,存在跨膜结构。荧光定量PCR检测结果表明,低温、高温和盐胁迫均可诱导Ca CBF1A基因的表达,其表达量在迅速达到峰值后又降低,说明Ca CBF1A是一个逆境胁迫快速响应基因,推测其在辣椒抗逆机制中起着重要的作用。

To reveal the roles of Ca CBF1 A gene played in pepper,the Ca CBF1 A gene was cloned and analyzed. According to the CBF gene sequence from Capsicum annuum genome,the primer was designed.Yujiao 101 was taken as material and Ca CBF1 A was acquired from the genome of pepper by PCR. The bioinformatics analysis results indicated that the gene possessed complete ORF with length of 471 bp,encoding 156 amino acids. Protein encoded by Ca CBF1 A gene in pepper contained conservative AP2 DNA binding domain. Besides,the subcellular localization and membrane structure were analyzed.It was predicted that protein encoded by Ca CBF1 A was located in chloroplasts,and contained transmembrane structure.Furthermore,the results by fluorescence quantitative PCR showed that cold,high temperature and salt stress could induce the expression of Ca CBF1 A gene,the expression level of which was decreased rapidly after peak point. These could provide some indication that Ca CBF1 A was rapid response gene,which played important roles in the resistance mechanisms of pepper.