کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
8359697 1542314 2016 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Transformation of Escherichia coli and protein expression using lipoplex mimicry
ترجمه فارسی عنوان
ترشح اشرشیا کولی و بیان پروتئین با استفاده از الگوریتم لیپولکس
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
چکیده انگلیسی
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 × 106 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.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 127, November 2016, Pages 68-72
نویسندگان
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