The Use of Protein S Fusion for Protein Solubilization
AssigneeUniversity of Medicine and Dentistry of New Jersey
InventorsMasayori Inouye, Takeshi Yoshida
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Abstract
The invention provides vectors containing a multiple cloning site comprising a PrS tag or a PrS2 tag from Myxococcus xanthus. Methods are provided for enhancing solubility of a target protein using Protein S tagged target fusion proteins.
Claims37
A vector containing a multiple cloning site comprising a nucleic acid sequence encoding a PrS tag or a PrS2 tag from Myxococcus xanthus.
A method for enhancing solubility of a target protein, comprising:
- a) fusing a nucleic acid sequence encoding at least one N-terminal domain of Protein S (PrS tag) from Myxococcus xanthus to a nucleic acid sequence encoding the target protein to create a nucleic acid sequence encoding a PrS tagged target protein;
- b) positioning the Protein S tagged target protein of step (a) into a vector;
- c) transforming a host cell with the vector;
- and
- d) culturing the host cell under conditions suitable for gene expression,
- whereby the expressed PrS tagged target protein is more soluble than the untagged target protein.
A PrS-tagged protein molecule bound to one or more myxospores of Myxococcus xanthus.
A method for purifying a PrS-tagged or PrS2-tagged protein comprising contacting the tagged protein with an affinity resin comprising myxospores of Myxococcus xanthus, thereby immobilizing the PrS-tagged or PrS2-tagged protein on the affinity resin.
A method of preparing a target protein for an analytical study, comprising preparing a PrS- or PrS2-tagged target protein, and performing the analytical study.
A method of characterizing a target protein comprising fusing at least one PrS tag or a PrS2 tag to the target protein, performing nuclear magnetic resonance spectroscopy and analyzing data from the nuclear magnetic resonance spectroscopy, thereby characterizing the target protein.
Dependencies are inferred from the claim text.