2010•Animal husbandry and feed scienceRequires access
Expression of recombinant bovine prion protein PrP27-30 in CHO-K1 cells.
Yaozhong Ding, Lina Ma, Zhong Jie, Yongsheng Liu
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Abstract
[Objective] To investigate the possibilities of expressing bovine PrP27-30 gene in CHO-K1 cells.[Method] The purified PCR products of PrP27-30 were digested and ligated to the pCI-neo vector to yield pCI-neo-PrP27-30 that was used as an expression vector.Then CHO-K1 cells were transfected by pCI-neo-PrP27-30,and stable expression clone cells were screened by methotrexate(MTX) at a concentration of 0.1 and 1.0 μmol/L.The transient expression was detected by indirect immunofluorescence assay and western blot.[Result] After drug selection with MTX,the expression of PrP27-30 gene was detected in CHO-K1 cells.[Conclusion] Recombinant protein PrP27-30 expressed in CHO-K1 cells has better immunoreactivity and can be used to study secondary structure and regulation mechanism of pathological isoform of prion protein(PrPC).