Author: Lu, Shuai; Chen, Yingzhu; Qin, Kun; Zhou, Jianfang; Lou, Yongliang; Tan, Wenjie
Title: Genetic and antigenic characterization of recombinant nucleocapsid proteins derived from canine coronavirus and canine respiratory coronavirus in China Document date: 2016_4_15
ID: 1ghgd7yg_10
Snippet: Both recombinant NPs of CCoV and CRCoV were ex-pressed in either soluble form or inclusion bodies, with obvious bands at about 45 and 55 kD respectively, which corresponds to our prediction ( Figure 3) . Optimization of the expression conditions indicated that N proteins had the highest soluble expression level with induction at 37°C for 4 h (data not shown). The soluble rNPs were purified by Ni 2+ and S75 columns. SDS-PAGE showed the isolated C.....
Document: Both recombinant NPs of CCoV and CRCoV were ex-pressed in either soluble form or inclusion bodies, with obvious bands at about 45 and 55 kD respectively, which corresponds to our prediction ( Figure 3) . Optimization of the expression conditions indicated that N proteins had the highest soluble expression level with induction at 37°C for 4 h (data not shown). The soluble rNPs were purified by Ni 2+ and S75 columns. SDS-PAGE showed the isolated CCoV-N and CRCoV-N both had a high purity of over 95%. Meanwhile, Western blot analysis demonstrated reactivity with His monoclonal antibody specifically at the corresponding sizes (Figure 4 ).
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