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高等植物光合同化产物蔗糖的质外体运输主要是靠蔗糖载体蛋白来完成的。MdSUT1是从苹果果实中克隆的蔗糖载体家族基因,本文将MdSUT1构建到酵母表达载体pMETαB中,重组质粒转化毕赤酵母PMAD16经0.5%甲醇诱导后获得Md-SUT1表达。纯化的MdSUT1异源表达蛋白免疫Balb/C小鼠制备多克隆抗体,抗体特异性分析结果显示该抗体对酵母表达和苹果中的MdSUT1识别具有较高的特异性。免疫共沉淀实验结果也证明抗体能够应用于苹果果实中MdSUT1的功能分析。
Higher plant photoassimilation product sucrose apoplast transport is mainly by sucrose carrier protein to complete. MdSUT1 is a family of sucrose vector cloned from apple fruit. MdSUT1 was constructed into yeast expression vector pMETαB. Md-SUT1 expression was induced after transformation of Pichia pastoris PMAD16 with 0.5% methanol. The purified MdSUT1 heterologous protein was used to immunize Balb / C mice to prepare polyclonal antibody. The results of antibody specificity analysis showed that the antibody has high specificity for yeast expression and MdSUT1 recognition in apple. Co-immunoprecipitation results also demonstrated that the antibody can be applied to the functional analysis of MdSUT1 in apple fruit.