Cell-Penetrating Peptide-Mediated Transformation of Large Plasmid DNA into Escherichia coli
Cell-Penetrating Peptide-Mediated Transformation of Large Plasmid DNA into Escherichia coli
复制标题
细胞穿透肽介导的大质粒 DNA 转化至大肠杆菌
DOI：
10.1021/acssynbio.9b00055
复制
发表时间：
2019
期刊：
影响因子：
4.7
通讯作者：
M. and Numata K.
中科院分区：
文献类型：
--
作者：
Islam M.M.;Odahara M.;Yoshizumi T.;Oikawa K.;Kimura M.;Su'etsugu;M. and Numata K.
细胞的高效遗传转化对于合成生物学过程是必不可少的，特别是对于大基因簇的转化。在这篇技术报告中，我们提出了一种新的细胞穿透肽（CPP）介导的大肠杆菌大尺寸质粒DNA转化系统。将含有CPP基序的阳离子肽与大质粒pMSR227（205kb）复合，成功地转化到E.大肠杆菌含有质粒DNA的转化体表现出编码红色荧光蛋白的报道基因的表达。转化效率显著高于热激法，与电穿孔法相近。该技术可用作在温和条件下简单高效转化大DNA分子的平台，而不会对DNA造成重大损害，加速了用于工业目的的基因工程微生物设计的合成生物学研究。
The highly efficient genetic transformation of cells is essential for synthetic biology procedures, especially for the transformation of large gene clusters. In this technical note, we present a novel cell-penetrating peptide (CPP)-mediated large-sized plasmid DNA transformation system forEscherichia coli. A large plasmid (pMSR227, 205 kb) was complexed with cationic peptides containing a CPP motif and was successfully transformed intoE. colicells. The transformants containing the plasmid DNA exhibited expression of a reporter gene encoding a red fluorescent protein. The transformation efficiency was significantly higher than that obtained using the heat-shock method and was similar to that of electroporation. This technique can be used as a platform for the simple and highly efficient transformation of large DNA molecules under mild conditions without causing significant damage to DNA, accelerating synthetic biology investigations for the design of genetically engineered microorganisms for industrial purposes.