Cell-Penetrating Peptide-Mediated Transformation of Large Plasmid DNA into Escherichia coli

Md Monirul Islam, Masaki Odahara, Takeshi Yoshizumi, Kazusato Oikawa, Mitsuhiro Kimura, Masayuki Su'Etsugu, Keiji Numata

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)

Abstract

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 for Escherichia coli. A large plasmid (pMSR227, 205 kb) was complexed with cationic peptides containing a CPP motif and was successfully transformed into E. coli cells. 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.

Original languageEnglish
Pages (from-to)1215-1218
Number of pages4
JournalACS Synthetic Biology
Volume8
Issue number5
DOIs
Publication statusPublished - May 17 2019
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

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