Optimizing Expression of Recombinant Collagen Peptide in Escherichia coli

Designed primers to obtain the target gene(CP6) fragment through PCR,the template contained the protein polypeptide gene of Ⅵ type human collagen,and constructed the recombinant plasmid,and then screened the host bacteria of high efficient expression,detected the instability of recombinant plasmid,as well as optimized the culture conditions.The results showed that the best host bacterium was Rosetta(DE3) and the recombinant plasmid has higher stability.The optimal expression conditions were inoculum of 4% and 5%,37 ℃,200 r/min,cultured to OD600 nm0.7 and added 0.5 mol/L IPTG to the culture medium,and then cultured 5 h.Western blotting showed that the expressed protein could specifically bind with human monoclonal antibody COL6A2.