西伯利亚蓼乙二醛酶Ⅱ基因的克隆与表达模式分析

Expression Modes Analysis and Cloning of GLY Gene from Polygonum sibiricum Laxm.

[目的]克隆西伯利亚蓼(Polygonum sibiricum Laxm.)乙二醛酶Ⅱ基因并了解该基因表达模式。[方法]应用RACE技术和实时荧光定量PCR技术从西伯利亚蓼中克隆了具有完整编码区的乙二醛酶Ⅱ基因的cDNA序列,并对该基因在不同胁迫时间、不同组织的表达差异进行了比较。[结果]克隆的乙二醛酶Ⅱ基因cDNA为890 bp,其中开放读码框为765 bp,编码254个氨基酸,5'非翻译区为49bp,3'非翻译区为72 bp,GenBank中登录号为HM241910。序列比对分析结果表明,该基因编码的乙二醛酶Ⅱ与乙二醛酶Ⅱ三型匹配最佳,并具备乙二醛酶Ⅱ的GloB、ComEC和Lactamase-B等结构域。实时荧光定量PCR分析显示,乙二醛酶Ⅱ基因在正常生长的西伯利亚蓼地下茎、茎与叶中都有表达,其中茎的表达量最高。同时,乙二醛酶Ⅱ基因受3%NaHCO3胁迫诱导,其在不同部位的表达模式有差异。[结论]克隆了西伯利亚蓼乙二醛酶Ⅱ基因并初步阐明了其表达模式。

[ Objective j The aim was to clone GLY gene from Polygonum sibiricum Laxm. and understand the expression modes of gene. [Meth-od] The full length of glyoxalase Ⅱ gene from Polygonum sibiricum kaxm. was isolated using RACE（ rapid amplification of cDNA ends） method. RT-PCR（ Real-time quantitative PCR） was used to research the expression of PsGLY-3 from different tissues and different deal time. [ Result] The full length of glyoxalase Ⅱ gene from Polygonum sibiricum Laxm. （GenBank accession：HM241910） was isolated using RACE（rapid amplification of cDNA ends） method,which extended 890 bp with a 49 bp 5＇ untranslated region, a 765 bp open reading frame （ORF） encoding a peptide of 254 amino acid residues, and a 72 bp 3＇ untranslated region. The glyoxalase Ⅱ had high similarity to GLY3. Similar with most of glyoxalase Ⅱ protein, glyoxalase Ⅱ from Polygonum sibiricum Laxm. include GloB domain, ComEC domain, Lactamase.B domain, etc.. Real-time quantitative PCR revealed that glyoxalase II gene was expressed in leaves, stem and rhizoma. The highest glyoxalase II gene expression was detected in stem. Moreover,glyoxalase II gene could be induced by 3 % NaHCO3, but the dynamics of the inductions varied in different tissues. [ Conclusion ] The research have cloned GLY zene from Polygonum sibiricum Laxm. and expounded the expression modes of PsGLY-3.