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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g DNA序列长1 954bp,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(g6pdh ID),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(g6pdh IR),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g6pdh IR在草菇同核体与异核体中的表达量均很低,因此转录本g6pdh ID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。
Glucose-6-phosphate dehydrogenase is the rate-limiting enzyme in the pentose phosphate pathway, affecting the production of NADPH required for cell life activities. In this study, two transcripts of this gene were cloned from Agrobacterium tumefaciens and their expression levels in two homokaryons and heteronuclear isolates were determined. The results showed that the g DNA sequence of g6pdh was 1 954 bp in length and contained 7 introns. Two transcripts could be produced: one was the full-cut transcript of intron (g6pdh ID), encoding a protein of 515 amino acids with an entire domain; the other is an alternative splice variant (g6pdh IR) retained by the fifth intron predicted for a protein encoding 316 amino acids with no complete domain . Quantitative PCR results showed that the g6pdh IR variable splicing variant was found to be low in both syncytium and heterokaryon, so the transcript g6pdh ID was the major splice body of g6pdh gene. The g6pdh gene was highly expressed The amount of expression in heterokaryons is much higher than in either of the two congeners that are weaker in growth. The conclusion of the research shows that the activity of carbohydrate metabolism is closely related to the growth and development of edible fungi.