甘蔗细胞质型苹果酸脱氢酶基因的克隆及其表达分析

Cloning and Expression Analysis of Cytosolic Malate Dehydrogenase Gene from Sugarcane

对甘蔗(Saccharum officinarum L.)叶片全长cDNA文库进行测序,获得了1个细胞质型苹果酸脱氢酶(cMDH)基因的全长cDNA序列,命名为Sc-cMDH。生物信息学分析表明,该基因全长1314 bp,开放阅读框为999 bp,编码332个氨基酸。Sc-cMDH与其他植物cMDH的氨基酸序列同源性高达86.5%～97.0%。Sc-cMDH包含典型的NAD+结合基元T11GAAGQI17和催化基元I184WGNH188,还有相当保守的6个半胱氨酸残基,因此推断该基因为细胞质型NAD-MDH。定量PCR分析结果表明,该基因在甘蔗叶片和根中的表达量高于茎。

A full-length cDNA sequence of sugarcane （Saccharum officinarum L.） cytosolic malate dehydrogenase （cMDH） gene, named as Sc-cMDH, was obtained from sugarcane full-length cDNA library by EST sequencing and bioinformatics analysis. The full-length of Sc-cMDH was 1314 bp with 999 bp open reading frame, encoding 332 amino acids. Sc-cMDH shared 86.5%-97.0% of amino acids with cMDH from other 9 species. Sc-cMDH contains typical NAD＋ binding motif （THGAAGQ117） and catalytic motif （II84WGNH188）. Real-time PCR analysis showed that the expression of Sc-cMDH was higher in leaves and roots than that in stems of sugarcane.