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US 2006/0216698 A1Published application
Fusion expression vector containing the His-tagged Vitreoscilla hemoglobin coding gene
OwnerKyung-Won Parkinventor
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OwnerKyung-Won Parkinventor
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The present invention relates to method of providing color to aid in the expression and purification of proteins of E. coli. Vector according to the present invention includes the His-tagged Vitreoscilla hemoglobin (VHb) coding gene upstream of the multiple cloning site. The vector was designed to express target proteins as VHb fusions, which can be purified in one step by affinity chromatography. Due to the color of the heme in VHb, the VHb-fused target proteins have a red color that provides a visual aid for estimating their expression level and solubility. The red color can also be used as a visual marker throughout purification, while the concentration of the fusion protein can be determined by measuring the amount of VHb using carbon monoxide difference spectra. In addition, because of inherently high solubility of VHb, the fusion can increase the solubility of sparingly soluble target proteins. Target proteins can be easily separated from His-tagged VHb due to the presence of a thrombin cleavage site between them.
A DNA vector comprising a fusion protein gene, wherein the fusion protein gene comprises a gene encoding at least 2 consecutive histidines, a globin gene or a gene encoding a fluorescent protein, a gene for a protease recognition site, a multiple cloning site, and a gene for a glycine rich linker.
A process for expression and purification of a fusion protein encoded by a DNA vector comprising a target protein gene, and a globin gene or a gene encoding a fluorescent protein, wherein the expressed fusion protein produces a color and wherein said color aids the purification of the target protein.
A fusion protein comprising a target protein and an end segment wherein said fusion protein further comprises at least 2 consecutive histidines, a globin protein, a protease recognition site, and a glycine rich linker.
Dependencies are inferred from the claim text.