Patent ID: 9714274
Date: 2017-07-25
CPC Classifications: C07K

Claim:
1. A method for enhancing expression of a fusion protein that comprises a target peptide, the method comprising culturing a host cell transformed with an expression vector comprising a nucleic acid encoding a fusion protein that comprises a fusion carrier protein linked to at least one target peptide, wherein the fusion carrier protein has an amino add sequence as set forth in Formula wherein T1is absent, a Met, a His-tag or at least one peptidic cleavage site, A1 is superfolder green fluorescent protein, which has the amino acid sequence Ser-Lys-Gly-Glu-Glu-Leu-Phe-Thr-Gly-Val-Val-Pro-lle-Leu-Val-Glu-Leu-Asp-Gly-Asp-Val-Asn-Gly-His-Lys-Phe-Ser-Val-Arg-Gly-Glu-Gly-Glu-Gly-Asp-Ala-Thr-Asn-Gly-Lys-Leu-Thr-Leu-Lys-Phe-lle-Cys-Thr-Thr-Gly-Lys- Leu-Pro-Val-Pro-Trp-Pro-Thr-Leu-Val-Thr-Thr-Leu-Thr-Tyr-Gly-Val-Gln-Cys-Phe-Ser-Arg-Tyr-Pro-Asp-His-Met-Lys-Arg-His-Asp-Phe-Phe-Lys-Ser-Ala-Met-Pro-Glu-Gly-Tyr-Val-Gln-Glu-Arg-Thr-lle-Ser-Phe-Lys-Asp-Asp- Gly-Thr-Tyr-Lys-Thr-Arg-Ala-Glu-Val-Lys-Phe-Glu-Gly-Asp-Thr-Leu-Val-Asn-Arg-lle-Glu-Leu-Lys-Gly-lle-Asp-Phe-Lys-Glu-Asp-Gly-Asn-lle-Leu-Gly-His-Lys-Leu-Glu-Tyr-Asn-Phe-Asn-Ser-His-Asn-Val-Tyr-lle-Thr-Ala- Asp-Lys-Gln-Lys-Asn-Gly-lle-Lys-Ala-Asn-Phe-Lys-lle-Arg-His-Asn-Val-Glu-Asp-Gly-Ser-Val-Gln-Leu-Ala-Asp-His-Tyr-Gln-Gln-Asn-Thr-Pro-lle-Gly-Asp-Gly-Pro-Val-Leu-Leu-Pro-Asp-Asn-His-Tyr-Leu-Ser-Thr-Gln-Ser- Val-Leu-Ser-Lys-Asp-Pro-Asn-Glu-Lys-Arg-Asp-His-Met-Val-Leu-Leu-Glu-Phe-Val-Thr-Ala-Ala-Gly-lle-Thr-His-Gly-Met-Asp-Glu-Leu-Tyr-Lys (SEQ ID NO: 1), or an amino acid sequence that is at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO: 1, T2 is absent, a His-tag or at least one peptidic cleavage site, provided that at most one of T1 and T2 is absent, and wherein the target peptide is selected from the group consisting of PTH, GLP-1, exenatide, liraglutide, calcitonin, and proinsulin, under suitable conditions for expression of the expression vector, thereby producing the fusion protein encoded by the nucleic acid in bacterial inclusion bodies, wherein the suitable conditions comprist an inducer for inducing the host cell to express the expression vector.