E.coli K-12琥珀酸脱氢酶sdhC基因纳米锥的设计及自组装

Self-assemble of gene nanocone with sdhC DNA sequence of E. coli K-12

采用DAEDALUS软件,以E.coliK-12MG1655的琥珀酸脱氢酶复合体C亚基编码酶基因sdhC的双链DNA为脚手架链,进行一个边长为17.68nm的正四面锥设计,并最终通过核酸自组装反应获得了sdhC基因的纳米锥聚合体。利用琼脂糖凝胶电泳、扫描电子显微镜和透射电子显微镜对多组样品进行化学组分和形貌分析,同时利用原子力显微镜液下成像法测定了核酸聚合体的三维尺度。结果表明:DAEDALUS软件设计出的16条订书钉链,确实将624bp的sdhC双链DNA中的一条链折叠成了平均边长为19.05nm的正四面锥,与预设模型仅相差1.37nm,证明普通基因的双链DNA可以替代M13mp18单链核酸作为DNA折纸的核酸材料,为构建核酸纳米材料提供了一种新方法。

E.coli K-12 MG1655 succinate dehydrogenase gene,sdhC(encoding membrane protein sdhC),a double-stranded DNA,was used as a scaffold to design a 17.68nm tetrahedron with DAEDALUS software.The sdhC DNA strand self-assembled into a nanocone polymer exactly as similar as the front design.The morphology,structure and chemical composition of the samples from several experiments were assayed by agarose gel electrophoresis,scanning electron microscope and transmission electron microscopy,at the same time,the three-dimensional structure was imaged by high-resolution atomic force microscope in liquid.The results showed 16 staple strands designed by DAEDALUS folded the 624 bp double-stranded DNA of sdhC gene into nanocones with an average length of 19.05nm,which is only 1.37nm difference with the prediction.This study provides a new method for constructing nucleic acid nanomaterial with common genetic sequences beside M13mp18 in DNA origami.