SsrA peptide tag from Mycoplasma fl orum has been developed as a versatile biotechnology tool to control orthogonal degradation of tagged proteins in Escherichia coli . Here, using the systematic deletion mutants of m...SsrA peptide tag from Mycoplasma fl orum has been developed as a versatile biotechnology tool to control orthogonal degradation of tagged proteins in Escherichia coli . Here, using the systematic deletion mutants of mf -ssrA tag, we demonstrated that the residues in two separate regions have diff erent functions in mf -Lon-mediated specifi c orthogonal target protein degradation in E. coli . The deletion of multiple residues, up to six amino acids, did not fatally abolish its specifi c degradation activity, instead of being able to improve the stability of the tagged protein in the presence of endogenous proteases before mf -Lon expression in E. coli . Except for previously identifi ed essential residues, the region adjacent to the C-terminal of the mf -ssrA tag was involved in mf -Lon and endogenous protease-mediated degradation. Moreover, the deletion of specifi c residues made the mf -ssrA tag more eff ective and compact. The mf -ssrA tag can be implemented in synthetic biology and bioengineering for development of synthetic circuits.展开更多
基金supported by the National Natural Science Foundation of China (No. 21476167 and No. 21778039)
文摘SsrA peptide tag from Mycoplasma fl orum has been developed as a versatile biotechnology tool to control orthogonal degradation of tagged proteins in Escherichia coli . Here, using the systematic deletion mutants of mf -ssrA tag, we demonstrated that the residues in two separate regions have diff erent functions in mf -Lon-mediated specifi c orthogonal target protein degradation in E. coli . The deletion of multiple residues, up to six amino acids, did not fatally abolish its specifi c degradation activity, instead of being able to improve the stability of the tagged protein in the presence of endogenous proteases before mf -Lon expression in E. coli . Except for previously identifi ed essential residues, the region adjacent to the C-terminal of the mf -ssrA tag was involved in mf -Lon and endogenous protease-mediated degradation. Moreover, the deletion of specifi c residues made the mf -ssrA tag more eff ective and compact. The mf -ssrA tag can be implemented in synthetic biology and bioengineering for development of synthetic circuits.
