Study On The Expression Of Recombinant Antimicrobial Peptides Mediated By Inclusion Body

In recent years,people had a new understanding of inclusion bodies for their biological function and the value of potential application.For cytotoxic proteins or unstable structure of short peptides,using the form of recombinant expression in inclusion bodies have showed unique natural advantage.However,the method is relatively single for inducing the formation of inclusion bodies in recombinant proteins,using the fusion tags that are easy to form inclusion bodies mediated the target protein for the formation of inclusion bodies at mos of time.In this essay,it was constructed a series of intracellular and secretory expression vectors of fusion proteins.In commonly,TrxA as a fusion tag that used to enhance the soluble expression of the target protein and the fusion protein is obviously accumulated in a soluble form in E.coli cells when fused with pEGF,Metch,or Andropin;However,it caused the formation of much inclusion bodies when the signal peptide of OmpA or DsbC was linked to the N-terminus of the fusion protein in induced expression,also the lower expression temperature was benifit for the accumulation of inclusion bodies.This result shows that the signal peptide can induce the formation of inclusion bodies of recombinant proteins in some of degree.This new discovery provides a new approach for the expression of recombinant proteins based on inclusion bodies.It was the unique antibacterial mechanisms that antimicrobial peptides shows the value of potential application.Exploring efficient and simple recombinant expression methods of antimicrobial peptides has important theoretical and practical significance.In this essay,the signal peptide was used to induce the formation of recombinant protein inclusion bodies,and successfully performed the highly efficient recombinant expression of the antimicrobialpeptides based on inclusion bodies in E.coli.Compared with the corresponding intracellular soluble expression,the inclusion bodies of antimicrobial peptides increased about 50%.It was a highly purity of inclusion bodies and the content of recombinant protein was more than 70% isolated from it.This research also established the technology system for separated and dissoluted the inclusion bodies and the corresponding TEV proteinase cleavaged recombinant protein for released antimicrobial peptide.The test result shows that the recombinant antibacterial peptides had significant antibacterial activity.Compared with reported methods for the expression and purification of antibacterial peptides,the established technical system had a high expression level and the purification method was more simpler and efficient,with a view to providing new ideas and new methods for the large-scale industrial production of antimicrobial peptides,which will in turn promote the development and practical application of antimicrobial peptides.