Molecular cloning and expression of a serine protease family from Jellyfish Cyanea capillata / 中国生化药物杂志

ABSTRACT

Objective To obtain a single toxin component from the jellyfish Cyanea capillata and provide a foundation for the further study on bioactivity and function of the serine proteases from C.capillata.Methods Primers designed with restriction enzyme were used to amplify the coding region of cDNAs (CcSP1, CcSP2 and CcSP3).PCR fragments were ligated with the pET-24a( +) vector to construct the recombinant plasmids (pET24a-CcSP1, pET24a-CcSP2 and pET24a-CcSP3).After screening and identification,the recombinant plasmids were transformed into the Rosetta (DE3).plysS for protein expression.After induction with IPTG, SDS-PAGE and Western-blot were used to detect the expression of the recombinant proteins.Results SDS-PAGE showed that the proteins of rCcSP1, rCcSP2 and rCcSP3 were expressed in a single band at about 34 kDa, 42 kDa and 42kDa, respectively.Western-blot detection with anti-His antibody further confirmed that these recombinant proteins were His-tagged CcSP1, CcSP2 and CcSP3 fusion protein were obtained.Conclusion Prokaryotic recombinant plasmids of C.capillata serine proteases are contructed and recombinant proteins are obtained, which establishes the foundation for future study on the function of serine proteases from jellyfish.