Renaturation method of COVID-19 recombinant protein inclusion bodies
A coronavirus and recombinant protein technology, applied in the biological field, can solve problems such as low recovery rate, large amount of protein precipitation, shortening the time and cost required for renaturation of recombinant protein inclusion bodies, etc.
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Embodiment 1
[0042] Example 1 Novel coronavirus RBD recombinant protein expression and inclusion body renaturation
[0043] 1. Expression of novel coronavirus RBD recombinant protein
[0044] (1) A BamH I restriction site is introduced into the N-terminus of the novel coronavirus RBD protein, and an Xho I restriction site is introduced into the C-terminus to obtain the novel coronavirus RBD recombinant protein. According to the gene sequence encoding the RBD recombinant protein of the new coronavirus, synthetic primers were designed for PCR amplification. After the PCR amplification product was purified, the PCR amplification product of the gene encoding the RBD recombinant protein of the new coronavirus was obtained. Among them, the full gene sequence of the novel coronavirus RBD protein is shown in SEQ ID NO: 1, the upstream primer for PCR amplification is: acacggatccAGAGTCCAACCAACAGAATC (SEQ ID NO: 3), and the downstream primer is: agagctcgagGAAATTGACACATTTGTTTTTAACC (SEQ ID NO: 4).
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Embodiment 2
[0059] Example 2 Expression of novel coronavirus N recombinant protein and renaturation of inclusion bodies
[0060] Including the following steps:
[0061] (1) A Sac I restriction site was introduced into the N-terminus of the N protein of the novel coronavirus, and 6*His and Not I restriction sites were introduced into the C-terminus to obtain the recombinant protein of the novel coronavirus N. According to the gene sequence encoding the new coronavirus N recombinant protein, synthetic primers are designed for PCR amplification. After the PCR amplification product is purified, the PCR amplification product of the coding gene of the new coronavirus N recombinant protein is obtained. Among them, the new coronavirus N protein The whole gene sequence is shown in SEQ ID NO:2, the upstream primer for PCR amplification is: tgtcgagctcATGTCTGATAATGGACCC (SEQ ID NO:5), and the downstream primer is: gagtgcggccgcTTAGTGATGGTGGTGATGGTGGGCCTGAGTTGAGTCAGC (SEQ ID NO:6).
[0062] (2) The PC...
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