东亚砂藓甘油醛-3-磷酸脱氢酶基因的克隆及表达分析

目的]克隆东亚砂藓甘油醛-3-磷酸脱氢酶基因RjGAPDH,并对其进行生物信息学和表达分析。方法]通过分析东亚砂藓转录组测序数据,利用RT-PCR技术克隆该基因的全长序列,并通过荧光定量PCR分析RjGAPDH基因在不同干旱胁迫时间的表达情况。结果]该基因全长为1 208 bp,开放阅读框1 053 bp,编码350个氨基酸,预测蛋白分子量为38.66 kD,等电点pI为6.02,不稳定系数为23.97,为稳定蛋白;该基因编码的蛋白无跨膜区和信号肽序列,定位于细胞质。在快速干旱胁迫处理过程中,东亚砂藓RjGAPDH基因的表达量高于正常生长的材料(CK),能被诱导表达。结论]RjGAPDH基因可能参与东亚砂藓对干旱的胁迫反应,为后续进一步研究其功能特征奠定基础。

Cloning and Expression Analysis of Glyceraldehyde-3-Phosphate dehydrogenase Gene RjGAPDH in Racomitrium japonicum

Objective] The research aimed to clone glyceraldehyde-3-phosphate dehydrogenase gene RjGAPDH,and analyze its bioinformation and expression.Method] Based on RNA-Seq databases of Racomitrium japonicum,RjGAPDH gene was cloned by RT-PCR,and the expression of RjGAPDH in different times under quick dry was detected by real-time PCR.Result] The full length cDNA sequence of RjGAPDH gene was 1 208 bp in length,and contained a 1 053 bp open reading frame which encoded a protein of 350 amino acids,molecular weight was 38.66 kD and theoretical pI was 6.02.The predicted RjGAPDH protein belonged to stable protein,located in cytoplasm without transmembrane protein and signal peptide.In the proceed of quick dry,RjGAPDH gene could be induced,and the expression level was higher than that in CK.Conclusion] RjGAPDH gene was speculated to participate the stress reaction of Racomitrumjaponium,and the results laid the foundations for the further study on its function.