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菌物学报  2015 

草菇葡萄糖-6-磷酸脱氢酶基因及其可变剪接体的克隆与表达量分析

DOI: 10.13346/j.mycosystema.150034, PP. 724-733

Keywords: 食用真菌,戊糖磷酸途径,基因克隆,定量PCR

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Abstract:

葡萄糖-6-磷酸脱氢酶是戊糖磷酸途径的限速酶,影响着细胞生命活动所需要的NADPH的产生。本研究从草菇中克隆到该基因的2个转录本,并测定了它们在两个同核体和形成的异核体菌株中的表达量。结果表明,草菇葡萄糖-6-磷酸脱氢酶基因(g6pdh)的gDNA序列长1954bp,有7个内含子,可产生两个转录本:一个是内含子全部剪切的转录本(g6pdhID),编码515个氨基酸且有完整结构域的蛋白质;另一个是第5个内含子保留的可变剪接变体(g6pdhIR),预测编码316个氨基酸但没有完整结构域的蛋白质。定量PCR结果显示,可变剪接变体g6pdhIR在草菇同核体与异核体中的表达量均很低,因此转录本g6pdhID为g6pdh基因的主要剪接体;并且g6pdh基因在生长旺盛的异核体中的表达量远远高于在生长较弱的两个同核体中的任何一个。研究结论说明,糖代谢活动的强弱与食用真菌的生长与发育有密切关系。

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