Effects of Mutations in IQ Motif of AtIQM1 on Its Calmodulin Binding

doi:10.13560/j.cnki.biotech.bull.1985.2014.12.021

Abstract

Abstract: To confirm the effects of mutation of key amino acid residues in IQ motif of AtIQM1 on its calmodulin binding, the LQ was deleted or L was mutated to S through the mutagenesis technology in vitro. Then calmodulin binding of the mutant protein iqm1 was analyzed via yeast two-hybrid assay. The self-activation was not observed when the recombinant plasmid pGADT7-IQM1CDS, pGADT7-IQM1Del113-114, pGBKT7- CaM5 or pGADT7-IQM1L113S was transformed into the yeast AH109 strain, respectively. Blue colonies were achieved on the selective agar plates (SD/-Ade/-His/-Leu/-Trp/X-α-Gal)when pGADT7-IQM1CDS and pGBKT7-CaM5 were co-transformed into AH109 yeast strain. However, the yeasts which were co-transformed with pGADT7-IQM1Del113-114 and pGBKT7-CaM5 or pGADT7-IQM1L113S and pGBKT7-CaM5 were not able to grow on the selective medium(SD/-Ade/-His/-Leu/-Trp). The results suggest that the LQ or L in IQ motif is required for AtIQM1 to combine with CaM5.

Key words: AtIQM1, Mutantion in IQ motif, Yeast two hybrid, Calmodulin binding

Huang Zhangke, Zhang Yi’neng, Mo Zhongzhen, Zhou Yuping, Huang Xiaoling, Tian Chang’en. Effects of Mutations in IQ Motif of AtIQM1 on Its Calmodulin Binding[J]. Biotechnology Bulletin, 2014, 0(12): 128-132.

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