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IQ基序突变对AtIQM1的钙调素结合活性的影响

DOI: 10.13560/j.cnki.biotech.bull.1985.2014.12.021, PP. 128-132

Keywords: AtIQM1,IQ,基序突变,酵母双杂交,CaM,结合

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Abstract:

旨在确定IQ基序中关键氨基酸残基突变对IQM1的CaM结合活性的影响,利用基因体外定点突变技术将LQ缺失或将L突变成S,并通过酵母双杂交分析突变蛋白iqm1的CaM结合活性。结果显示,用重组质粒pGADT7-IQM1CDS、pGADT7-IQM1Del113-114、pGADT7-IQM1L113S或pGBKT7-CaM5分别转化酵母AH109,未出现自激活现象;用pGADT7-IQM1CDS和pGBKT7-CaM5共转化酵母能在选择性培养基(SD/-Ade/-His/-Leu/-Trp/X-α-Gal)上形成蓝色菌落;而用pGADT7-IQM1Del113-114和pGBKT7-CaM5或pGADT7-IQM1L113S和pGBKT7-CaM5共转化酵母则不能在选择性培养基(SD/-Ade/-His/-Leu/-Trp)上形成菌落。结果表明,IQ基序中LQ或者L是IQM1与CaM5结合的关键氨基酸残基。

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