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US 10,239,922 B2Granted
Fusion proteins of superfolder green fluorescent protein and use thereof
AssigneeSuzhou Kunpeng Biotech Co., Ltd.
InventorsWenshe Liu, Wei Wan
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Abstract
The present disclosure pertains to methods of producing recombinant peptides that contain between 10 and 200 amino acid residues using novel carrier proteins derived from superfolder green fluorescent protein and its mutants.
Claims10
A method for enhancing expression of 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 the target peptide, wherein the fusion carrier protein has an amino acid sequence as set forth in Formula
- T1-A1-T2 (I),
- wherein
- T1 is absent, a Met, a His-tag, or at least one peptidic cleavage site,
- A1 is a superfolder green fluorescent protein, which has the amino acid sequence Ser-Lys-Gly-Glu-Glu-Leu-Phe-Thr-Gly-Val-Val-Pro-Ile-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-Ile-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-Ile-Ser-Phe-Lys-Asp-Asp-Gly-Thr-Tyr-Lys-Thr-Arg-Ala-Glu-Val-Lys-Phe-Glu-Gly-Asp-Thr-Leu-Val-Asn-Arg-Ile-Glu-Leu-Lys-Gly-Ile-Asp-Phe-Lys-Glu-Asp-Gly-Asn-Ile-Leu-Gly-His-Lys-Leu-Glu-Tyr-Asn-Phe-Asn-Ser-His-Asn-Val-Tyr-Ile-Thr-Ala-Asp-Lys-Gln-Lys-Asn-Gly-Ile-Lys-Ala-Asn-Phe-Lys-Ile-Arg-His-Asn-Val-Glu-Asp-Gly-Ser-Val-Gln-Leu-Ala-Asp-His-Tyr-Gln-Gln-Asn-Thr-Pro-Ile-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-Ile-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,
- 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 comprise an inducer for inducing the host cell to express the expression vector, and wherein the expression level of the target peptide is enhanced as compared to a control where a fusion carrier protein is not used.
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Dependencies are inferred from the claim text.