Abstract
High-level expression of many eukaryotic genes has proved difficult to achieve even when a strong promoter1–3 and the ribosome binding sequence4,5 from highly expressed Escherichia coli genes have been placed in front of the coding sequences. To overcome this problem, many eukaryotic proteins have been efficiently produced as hybrids after fusion of their genes with a coding sequence of E. coli genes6. However, such hybrid proteins are not suitable for functional studies or clinical use unless the authentic protein sequence can be released by specific cleavage. Here, we have inserted the sequence Ile-Glu-Gly-Arg between the 31 amino-terminal residues of λ cII protein and Val 1 of human β-globin, and produced this hybrid in high yield in E. coli. We then cleaved the hybrid specifically at the single arginine, using blood coagulation factor Xa and thus liberated the authentic β-globin chain. As factor Xa is specific for the tetrapeptide Ile-Glu-Gly-Arg7, which is rare in protein sequences, our expression/cleavage system is applicable to the efficient production of many eukaryotic proteins.
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Nagai, K., Thøgersen, H. Generation of β-globin by sequence-specific proteolysis of a hybrid protein produced in Escherichia coli. Nature 309, 810–812 (1984). https://doi.org/10.1038/309810a0
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DOI: https://doi.org/10.1038/309810a0
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