AGO蛋白在小RNA介导的基因沉默中发挥关键作用,AGO10和AGO7分别为AGO1和AGO2家族成员,为了进一步研究AGO7和AGO10的功能。在同实验室对玉米Chang 7-2 AGO7和AGO10基因的克隆和表达分析的基础上,经过IPTG诱导蛋白表达,Ni柱亲和层析纯化蛋白,同时免疫家兔制备AGO7和AGO10蛋白的抗体。结果表明：IPTG诱导重组蛋白表达,在分子量37.69kDa和 38.44kDa处出现重组蛋白,与预测的分子量大小一致。Western blot检测显示免疫家兔的抗血清与玉米的AGO家族蛋白杂交信号良好。成功制备出玉米AGO7和AGO10抗体,为进一步开展玉米AGO7及AGO10的功能研究奠定了良好的基础。
英文摘要:
AGO proteins play important roles in small RNA mediated gene silencing. AGO10 and AGO7 are the numbers of AGO1 and AGO2 family, respectively. In order to study the function of AGO7 and AGO10. Based on the analysis of cloning and expression of Chang 7-2 AGO7 and AGO10 genes in maize and induced by IPTG. Proteins were purified by Ni affinity chromatography. Antibodies of AGO7 and AGO10 were obtained by rabbit immunization. The conclusion showed: 37.69kDa and 38.44kDa recombinant protein of AGO7 and AGO10 were massively expressed and purified by Ni affinity chromatography. Western blot showed relatively specific hybridization signals, which indicated that the polyclonal antibodies of AGO7 and AGO10 wereprepared successfully. This work will laid a good foundation for further research of the functions of AGO7 and AGO10 proteins .
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