DNA origami technique, a breakthrough in DNA nanotechnology, has been widely used to prepare complex DNA nanostructures with nanoscale addressability. However, the purity and yield are generally the bottleneck to appl...DNA origami technique, a breakthrough in DNA nanotechnology, has been widely used to prepare complex DNA nanostructures with nanoscale addressability. However, the purity and yield are generally the bottleneck to application of DNA nanostructures, and current methods for purifying DNA origami nanostructures in large quantities are time-consuming and laborious. This study aims to develop a scalable, cost-effective and contamination-free method of purifying DNA origami nanostructures. We employ an effective and convenient purification approach to purify planar rectangle DNA origami structures through rate-zonal centrifugation. By subjecting DNA origami samples to high centrifugal force in a density gradient media of glycerol, well-folded nanostructures and by-products are separated successfully, which are confirmed by agarose gel electrophoresis and atomic force microscopy(AFM). This method will aid the production of pure rectangle DNA origami nanostructures in large quantity.展开更多
以大肠杆菌菌毛蛋白CsgA组装形成的蛋白纤维为模板,引导不同数目的DNA四面体(tetrahedron DNA nanostructure,TDN)组装构建了蛋白-DNA亚微米复合结构. TDN经次氮基三乙酸(NTA)修饰后在Ni2+的螯合作用下与CsgA蛋白单体结合,利用CsgA的自...以大肠杆菌菌毛蛋白CsgA组装形成的蛋白纤维为模板,引导不同数目的DNA四面体(tetrahedron DNA nanostructure,TDN)组装构建了蛋白-DNA亚微米复合结构. TDN经次氮基三乙酸(NTA)修饰后在Ni2+的螯合作用下与CsgA蛋白单体结合,利用CsgA的自组装能力将TDN有序地排列在形成的蛋白纤维上.原子力显微镜(atomic force microscopy,AFM)成像结果表明,控制TDN与CsgA的浓度比为1:500,可以得到单个TDN与蛋白纤维的组装产物.将2个TDN通过杂交形成二聚体(dTDN)与CsgA蛋白进行组装,得到的亚微米复合结构保持了很好的直链形态,在蛋白纤维上连有3个dTDN结构的比例达44%.展开更多
基金Supported by Shanghai Natural Science Foundation(Nos.15ZR1448400 and 15ZR1448700)National Natural Science Foundation of China(No.31300825)
文摘DNA origami technique, a breakthrough in DNA nanotechnology, has been widely used to prepare complex DNA nanostructures with nanoscale addressability. However, the purity and yield are generally the bottleneck to application of DNA nanostructures, and current methods for purifying DNA origami nanostructures in large quantities are time-consuming and laborious. This study aims to develop a scalable, cost-effective and contamination-free method of purifying DNA origami nanostructures. We employ an effective and convenient purification approach to purify planar rectangle DNA origami structures through rate-zonal centrifugation. By subjecting DNA origami samples to high centrifugal force in a density gradient media of glycerol, well-folded nanostructures and by-products are separated successfully, which are confirmed by agarose gel electrophoresis and atomic force microscopy(AFM). This method will aid the production of pure rectangle DNA origami nanostructures in large quantity.
文摘以大肠杆菌菌毛蛋白CsgA组装形成的蛋白纤维为模板,引导不同数目的DNA四面体(tetrahedron DNA nanostructure,TDN)组装构建了蛋白-DNA亚微米复合结构. TDN经次氮基三乙酸(NTA)修饰后在Ni2+的螯合作用下与CsgA蛋白单体结合,利用CsgA的自组装能力将TDN有序地排列在形成的蛋白纤维上.原子力显微镜(atomic force microscopy,AFM)成像结果表明,控制TDN与CsgA的浓度比为1:500,可以得到单个TDN与蛋白纤维的组装产物.将2个TDN通过杂交形成二聚体(dTDN)与CsgA蛋白进行组装,得到的亚微米复合结构保持了很好的直链形态,在蛋白纤维上连有3个dTDN结构的比例达44%.
