|کد مقاله||کد نشریه||سال انتشار||مقاله انگلیسی||ترجمه فارسی||نسخه تمام متن|
|1172896||961625||2016||3 صفحه PDF||سفارش دهید||دانلود کنید|
• Periplasmic expression of MBP-Hep25 is toxic for E. coli.
• Using this property, we designed a new positive-selection expression vector.
• Cloning efficiency with this vector was close to 100%.
Recombinant proteins are often produced in the periplasm of Escherichia coli because this facilitates the purification process. The oxidizing environment favors the formation of disulfide bridges. We showed that the periplasmic expression of the human hormone hepcidin 25 (Hep25) fused to the maltose-binding protein (MBP) resulted in cell death. This toxicity was not observed when MBP–Hep25 accumulated in the bacterial cytoplasm, or when Hep25 was addressed to the periplasm without the MBP tag. We then modified the periplasmic expression vector pMALp2E to create pMALp2EH, a positive-selection vector with Hep25 as counterselection gene.
Journal: Analytical Biochemistry - Volume 500, 1 May 2016, Pages 35–37