摘要
利用分子克隆手段构建了酿酒酵母ATP合酶δ亚基和流感病毒血凝素(haemagglutinin,HA)标签融合蛋白的表达质粒。该融合蛋白在酿酒酵母细胞中能够正常表达,而且能够互补编码δ亚基的ATP16基因缺失株在利用非发酵性碳源方面的表型缺陷,表明该融合蛋白具有野生型δ亚基的功能。同时,构建了在大肠杆菌细胞中表达该δ亚基的ScAtp16p-His6融合蛋白,并用纯化的融合蛋白在家兔中制备了其多抗血清。结果表明此多抗可以很好地与ScAtp16p-His6和HA-ScAtp16p两种融合蛋白特异性结合。这些研究材料的获得为深入研究ATP合酶的解偶联机制和磷酸化调控机理奠定了基础。
A construct expressing the HA-tagged δ subunit of F1F0 ATP synthase was constructed. Functional complementation study showed that the fusion protein could complement the function of the wild-type δ subunit in the utilization of non-fermentable carbon sources in yeast cells. Meanwhile, antiserum against inclusion bodies containing bacterial expressed ScATP16-His6 proteins was generated, and this polyclonal antibody could recognize fusion proteins ScATP16-His6 and HA-ScAtp16p. These studies provide a basis to further study the uncoupling mechanism between F1 and F0 portions of ATP syuthase and its regulation by protein phosphorylation in the mitochondrion.
出处
《生物技术通报》
CAS
CSCD
北大核心
2009年第8期99-103,共5页
Biotechnology Bulletin
基金
国家自然科学基金项目(30870107)
