东亚砂藓类萌发素蛋白基因RjGLP的克隆及表达分析  

Cloning and expression analysis on RjGLP, a germin-like protein gene from Racomitrium japonicum

作　　者：张梅娟[1] 沙伟[1,2] 刘博[2] 徐红红[2] 

机构地区：[1]东北林业大学生命科学学院,黑龙江哈尔滨150040 [2]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006

出　　处：《湖南农业大学学报（自然科学版）》2015年第1期47-52,66,共7页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金项目(31070180;31270254);黑龙江省自然科学基金重点项目(ZD201408)

摘　　要：以东亚砂藓转录组测序获得的类萌发素蛋白基因序列为基础,采用PCR技术克隆得到该基因的cDNA全长序列,命名为RjbZIP。该序列全长为726 bp,包含669 bp的开放阅读框,编码222个氨基酸的蛋白质。生物信息学分析显示,该蛋白的相对分子质量为23 397.9,等电点(pI)为6.82,不稳定系数为14.41,为稳定蛋白。序列预测分析表明,该蛋白疏水性强,具有信号肽序列,是一种胞外基质分泌蛋白。多序列比对分析结果显示,该基因编码的蛋白具有GLPs家族的典型特征。实时荧光定量PCR研究表明,在干旱胁迫处理过程中,东亚砂藓RjGLP基因的表达量高于正常生长的材料(CK),被诱导表达,推测该基因可能参与对干旱胁迫的应答。A full length cDNA sequence of germin-like protein was cloned from Racomitriumjaponicum using RT-PCR, and was named as RjGLP, which was 726 bp in length, contained an open reading frame （ORF） of 669 bp, and encoded with 222 amino acids. The results of bioinformatics demonstrated that the protein belonged to stable protein with molecular weight of 23 397.9 and wtih pI of 6.82. The sequence prediction showed that the protein, located in cytoplasm, was a transmembrane protein and had signal peptide. Multiple sequence alignment of GLP proteins revealed that the protein had typical characteristics of plant germin-like proteins. The RT-qPCR results showed that RjGLP gene could express in the process of quick dehydration, and the expression was higher than that of CK. Therefore, the RjGLP gene was speculated to participate in the stress reaction, and the results would lay the foundations for further study of desiccation-tolerant mechanism.

关 键 词：东亚砂藓 类萌发素蛋白 RjGLP基因 基因克隆 表达分析 

分 类 号：Q785[生物学—分子生物学]