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6-磷酸葡萄糖酸脱氢酶(6PGDH)是磷酸戊糖途径的限速酶,影响着细胞生命活动所需要的NADPH的产生.为揭示草菇菌丝生长的分子基础,通过克隆草菇6-磷酸葡萄糖酸脱氢酶编码基因(6gpdh)的可变剪接体,测序后进行生物信息学分析,利用表达谱和qPCR测定了它们在同核体和异核体菌株的表达量.结果显示,草菇6gpdh序列全长2 315bp,含10个内含子,第6个内含子存在内含子保留的可变剪接.在同核体和异核体菌株中,内含子保留的可变剪接体表达量低,且菌株间没显著差异,而内含子被剪接的剪接体表达量高,且异核体菌株高于同核体菌株.两种可变剪接体具有6-磷酸葡萄糖酸脱氢酶保守结构域和相似的三维结构.因此,初步分析两个可变剪切体是同工酶,内含子保留是组成型表达,表达量与菌丝生长快慢无关,内含子剪接是主效剪接方式,菌丝生长快的菌株表达量高,生长慢的菌株表达量低.图6表1参26
6-phosphogluconate dehydrogenase (6PGDH) is the rate-limiting enzyme in the pentose phosphate pathway, affecting the production of NADPH required by the cell life activities.In order to reveal the molecular basis of the growth of mushroom mycelium, Phosphogluconate dehydrogenase encoding gene (6gpdh) of the alternative splicing, sequenced bioinformatics analysis, using the expression profile and qPCR measured their homokaryotic and heterokaryotic strains of the expression.The results show that the grass The 6gpdh sequence of Mushroom contains a total length of 2 315 bp, contains 10 introns, and the 6th intron has an alternative splicing of the intron. In introns and heterokaryotic strains, alternative splicing The expression level was low and there was no significant difference among the isolates, while the intron was spliced and the heterokaryon strain was higher than that of the homokaryon splicing strain.The two alternative splicing fragments had 6-phosphogluconate off Hydrogenase conserved domain and similar three-dimensional structure.Therefore, preliminary analysis of the two variable splicing is an isozyme, intron retention is constitutive expression, expression and mycelium growth has nothing to do, intron splicing is The main effect of splicing, mycelial growth of the rapid expression of high strains, slow growth of the expression of strains Figure 1 reference 26 in Table 6