title: Transformation of Escherichia coli and protein expression using lipoplex mimicry.
abstract: We investigated a "one-step" method for transformation of and protein expression in Escherichia coli (E. coli) using a complex of n-stearylamine, a cationic lipid, and plasmid DNA, which mimics lipoplex-based approaches. When E. coli cells were treated with the cationic lipid-plasmid complex, the transformation efficiencies were in the range of approximately 2-3 × 10(6) colony-forming units. Further increase in the efficiency was obtained by co-treatment with calcium chloride (or rubidium chloride) and the complexes. Moreover, after DNA transfer, E. coli cells successfully expressed plasmid-encoded proteins such as cytochrome P450s and glutathione-S-transferase without overnight incubation of the cells to form colonies, an indispensable step in other bacterial transformation methods. In this study, we provide a simple method for E. coli transformation, which does not require the preparation of competent cells. The present method also shortens the overall procedures for transformation and gene expression in E. coli by omitting the colony-forming step.
authors.affiliation: School of Biological Sciences and Technology, Chonnam National University, Gwangju 61186, Republic of Korea.
authors.forename: Chul-Ho
authors.initials: CH
authors.lastname: Yun
authors.affiliation: College of Veterinary Medicine, Chonnam National University, Gwangju 61186, Republic of Korea.
authors.forename: Chun-Sik
authors.initials: CS
authors.lastname: Bae
authors.affiliation: College of Veterinary Medicine, Chonnam National University, Gwangju 61186, Republic of Korea. Electronic address: thahn@jnu.ac.kr.
authors.forename: Taeho
authors.initials: T
authors.lastname: Ahn
mesh_terms: D000588:Amines / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D010957:Plasmids / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014169:Transformation, Bacterial
chemical_list: D000588:Amines; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System; D005982:Glutathione Transferase; C009317:stearylamine
keywords: Escherichia coli; Expression; Lipoplex; Transformation; n-stearylamine