The method leading to overexpression of the full-length mouse recombinant prion protein (mrPrP 23-231) in the cytoplasm of E. coli as a his-PrP fusion protein and its effective purification using affinity chromatography is described. A typical yield of the method was 8-10 mg his-mrPrP per L of the bacterial culture. The purity of purified protein was > 95 %. The purified his-mrPrP was converted to a soluble form and its folding to alpha-helical and beta-sheet conformations was studied. The properties of differently folded mrPrP were determined by measuring their circular dichroism spectra, partial resistance to cleavage by proteinase K and by centrifugation in sucrose gradient.

• MeSH
• chromatografie afinitní MeSH
• cirkulární dichroismus MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• Escherichia coli genetika   metabolismus   MeSH
• myši MeSH
• oxidace-redukce MeSH
• peptidové fragmenty biosyntéza   chemie   genetika   izolace a purifikace   MeSH
• priony biosyntéza   chemie   genetika   izolace a purifikace   MeSH
• rekombinantní proteiny biosyntéza   chemie   genetika   izolace a purifikace   MeSH
• sbalování proteinů MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• peptidové fragmenty MeSH
• prion protein (23-231) MeSH Prohlížeč
• priony MeSH
• rekombinantní proteiny MeSH