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甘油醛-3-磷酸脱氢酶(glyceraldehyde-3-phosphatedehydrogenase,GAPDH)是真核生物细胞中一类功能丰富的蛋白质.采用抑制性差减杂交(suppressionsubtractivehybridization,SSH)的方法,筛选到一个在大豆疫霉侵染早期上调表达的、编码GAPDH的cDNA片段;克隆了该基因的全长序列,命名为PsGapdh.Southern杂交结果显示,PsGapdh在大豆疫霉基因组中含有3个拷贝.该基因编码的氨基酸序列具有GAPDH的保守结构域.在系统发育树中,PsGAPDH与两性绵霉(Achlyabisexualis)的GapC1同源性最高、与中华盒形藻(Odontellasinensis)和三角褐指藻(Phaeodactylumtricornutum)的GapC2同源性较高,三者聚类分布于GapC亚族的C-Ⅱ分支.RT-PCR分析表明,该基因在大豆疫霉侵染早期转录水平提高.该基因可以恢复酵母突变体?tdh1对H2O2的耐受性.上述结果提示,大豆疫霉的甘油醛-3-磷酸脱氢酶具有与酵母tdh1相似的抗氧化作用,且参与了大豆疫霉对寄主植物的侵染过程.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a kind of function-rich protein in eukaryotic cells.A method of suppression subtractive hybridization (SSH) The GAPDH-encoding cDNA fragment was up-regulated at the early stage of mildew inoculation, and the full-length sequence of the gene was cloned and named as PsGapdh.The results of Southern hybridization showed that PsGapdh contained 3 copies in Phytophthora sojae genome.The amino acid sequence Has a conserved domain of GAPDH.In the phylogenetic tree, PsGAPDH has the highest homology with GapC1 of Achlyabia xylani and GapC2 homology with Odontellasinensis and Phaeodactylum metricornutum High and the three clustered in the C-Ⅱ branch of GapC subfamily.RT-PCR analysis showed that the gene was increased in the early stage of Phytophthora sojae infection.The gene could restore the tolerance of yeast mutant? Tdh1 to H2O2 The above results suggest that glyphosate-3-phosphate dehydrogenase from Phytophthora sojae has antioxidant activity similar to that of yeast tdh1 and is involved in the host plantation of Phytophthora sojae The infection process.