小鼠重组朊蛋白核心片段PrP23-231的原核表达

Objective: To obtain highly expressed mature murine prion protein PrP23-231 and to understand the mechanism of prion protein conformational misfolding and amyloid fibrillization. Methods: Target gene fragment of PrP23-231 was obtained by PCR amplification from DNA genome of health C57BL/C mouse, then the purified target gene was cloned into a prokaryotic expression vector pET-30a, and thus recombinant pET-30a-PrP plasmid was constructed. The positive recombinant pET-30a-PrP plasmids were transformed into E.coli BL21, and IPTG was used to induce the expression of recombinant prion protein PrP23-231. Finally, SDS-PAGE and Western blotting identified the recombinant prion protein PrP23-231. Conclusion: SDS-PAGE and Western blotting showed that the obtained recombinant protein was exactly prion protein PrP23-231 with good immune characteristics. Conclusion: It indicated that the highly expressed recombinant prion protein PrP23-231 could be obtained by prokaryotic expression system, and the recombinant prion protein PrP23-231 could be provided as material for prion protein conformational misfolding and amyloid fibrillization.