衔接重复序列对酵母菌絮凝蛋白功能的调控作用
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国家自然科学基金(30970087)


Regulation of tandem repeats on the function of flocculation protein in Saccharomyces cerevisiae
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Supported by the National Natural Science Foundation of China (30970087)

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    摘要:

    摘要:【目的】了解絮凝基因FLO1 中重复DNA 序列B 和D 对絮凝蛋白Flo1p 功能的影响,为构建遗传稳定的最小絮凝功能基因奠定理论基础。【方法】通过PCR 和融合PCR 方法分别克隆到完整的絮凝基因FLO1、重复DNA 序列B 和D 分别缺失的衍生基因FLO1b 和FLO1d,分析这些基因在非絮凝酵母中表达对细胞絮凝特性的影响。【结果】与完整絮凝基因相比,重复DNA 序列B 和D 分别缺失后对酵母细胞絮凝强度没有明显影响,但不同基因在酵母菌中表达产生的絮凝特性受环境因素,如甘露糖浓度和pH 等的影响有明显差 异。FLO1 中重复DNA 序列B 和D 缺失后,细胞絮凝特性受甘露糖抑制的敏感性降低;同时对环境pH的改变具有更广泛的适应性。【结论】重复DNA 序列B 和D 对絮凝蛋白Flo1p 结构和功能具有调控作用,二者缺失后,特别是D 缺失后会使絮凝蛋白在极端酸碱环境下更稳定。

    Abstract:

    Abstract:[Objective]There are a large numbers of tandem repeats in FLO1,which are highly dynamic components in genome leading to the unstable flocculation profiles in Saccharomyces cerevisiae. The effects of repeated unite B or D deletion on the function of flocculation protein was studied to provide theory basis for constructing genetically stable flocculation gene with minimal size.[Methods]We cloned the intact flocculation gene FLO1 from S.cerevisiae YS59 by PCR,and constructed the derived genes FLO1b and FLO1d with repeated unite B or D deletion respectively by fusion PCR. We analyzed the physiological characteristics of flocculation in yeast strains YSF1,YSF1b and YSF1d containing FLO1,FLO1b and FLO1d respectively.[Results]YSF1b and YSF1d displayed almost the same level of Flo1-type flocculation as YSF1. However,flocculation of YSF1b and YSF1d,especially YSF1d was more tolerant to pH change and mannose concentration than strain YSF1.[Conclusion]Tandem repeats regulate the function of flocculation protein. Deletion of repeated unite B or D,especially D increases the stability of flocculation protein.

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李娥娥,常琦,郭雪娜,何秀萍,张博润. 衔接重复序列对酵母菌絮凝蛋白功能的调控作用. 微生物学报, 2012, 52(1): 69-76

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  • 收稿日期:2011-10-24
  • 最后修改日期:2011-11-25
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  • 在线发布日期: 2012-01-13
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