MODIFIED SAK GENE FOR THE PRODUCTION OF RECOMBINANT PROTEINS
First Claim
Patent Images
1. A process for the preparation of heterologous protein of interest in E. coli comprising the steps of:
- a) preparing a fusion DNA comprising a first DNA encoding modified SAK protein and a second DNA fused in the frame encoding the heterologous protein of interest,b) cloning of the fusion DNA of step a into an expression vector,c) expressing the fusion protein in E. coli cells,d) optionally refolding and purifying the fusion protein;
e) enterokinase cleavage of fusion protein, andf) isolating and purifying the protein of interest to obtain pure heterologous protein.
0 Assignments
0 Petitions
Accused Products
Abstract
The present invention relates to modified SAK gene having amino acid SEQ ID 2. The present invention further relates to process for cloning and expressing modified SAK gene fusion protein which imparts improved stability to the heterologous protein of interest. Further the invention relates to process of purification of recombinant heterologous proteins from bacterial inclusion bodies using modified SAK.
-
Citations
10 Claims
-
1. A process for the preparation of heterologous protein of interest in E. coli comprising the steps of:
-
a) preparing a fusion DNA comprising a first DNA encoding modified SAK protein and a second DNA fused in the frame encoding the heterologous protein of interest, b) cloning of the fusion DNA of step a into an expression vector, c) expressing the fusion protein in E. coli cells, d) optionally refolding and purifying the fusion protein; e) enterokinase cleavage of fusion protein, and f) isolating and purifying the protein of interest to obtain pure heterologous protein. - View Dependent Claims (2, 3, 4, 5)
-
-
6. A method for purification of modified SAK fusion protein wherein, the fusion protein is expressed as inclusion bodies in E coli comprising steps of:
-
a) solubilizing the inclusion bodies, b) refolding the solubilizing inclusion bodies, c) enterokinase digestion of modified SAK fusion protein, and d) isolating and purifying the protein of interest by one or more of chromatographic techniques. - View Dependent Claims (7, 8)
-
-
9. The method as claimed in 6, wherein ion exchange chromatography is selected from cation exchange and anion exchange in any order.
-
10. The method as claimed in 8, wherein hydrophobic interaction chromatography is performed using resins selected from butyl sepharose, phenyl sepharose or octyl sepharose.
Specification