Fusion protein containing a single-stranded dna binding protein and methods for expression and purification of the same
OwnerDaochun KONGinventor
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Abstract
The present invention provides an expression vector comprising a promoter and a polynucleotide sequence encoding a fusion protein. The present invention further provides a method for purification of an interest protein. The present invention also provides a fusion protein comprising a single-stranded DNA binding protein and an interest protein or polypeptide fused directly or indirectly with the COOH-terminus or NH2-terminus of the single-stranded DNA binding protein, wherein said fusion protein is capable of binding to single-stranded DNA.
Claims20
An expression vector comprising a promoter and a polynucleotide sequence encoding a fusion protein;
- wherein the fusion protein-encoding polynucleotide sequence is so operably linked to the promoter that the transcription of the fusion protein-encoding polynucleotide sequence is controlled by the promoter;
- wherein the fusion protein comprises a single-stranded DNA-binding protein and an interest protein or polypeptide fused directly or indirectly with the COOH-terminus or NH2-terminus of the single-stranded DNA-binding protein;
- and wherein the fusion protein is capable of binding to single-stranded DNA.
A method for purification of an interest protein, said method comprising:
- contacting a host cell with an expression vector, wherein the expression vector comprises a promoter and a polynucleotide sequence operably linked to the promoter, wherein the polynucleotide sequence encodes a fusion protein comprising a single-stranded DNA binding protein and the interest protein fused directly or indirectly with the COOH-terminus or NH2-terminus of the single-stranded DNA binding protein, and wherein the fusion protein is capable of binding to single-stranded DNA;
- culturing the host cell under such conditions that the fusion protein is expressed;
- lysing the host cell to obtain a cell lysate;
- contacting the cell lysate with a substrate immobilized with single-stranded DNA to allow the fusion protein to bind to the single-stranded DNA of the substrate;
- washing the substrate to remove impurities;
- and
- eluting the bound fusion protein from the substrate;
- thereby the interest protein is expressed and purified in the form of the fusion protein.
A fusion protein comprising a single-stranded DNA binding protein and an interest protein or polypeptide fused directly or indirectly with the COOH-terminus or NH2-terminus of the single-stranded DNA binding protein, wherein said fusion protein is capable of binding to single-stranded DNA;
- thereby the fusion protein can be purified by a substrate immobilized with single-stranded DNA.
Dependencies are inferred from the claim text.