Method of producing and purifying soluble recombinant coq5 protein and soluble recombinant coq5 protein thereof
AssigneeChang Gung University
InventorsHSIU-CHUAN YEN, SHIH-WEI CHEN
Life of this patent
Estimating the term…
Abstract
The present invention relates to a producing and purifying method of soluble recombinant COQ5 protein, which is expressed in soluble form by E. coli, under native conditions. The method is characterized by pre-treating bacterial lysate with low concentration of ionic detergent, such as SDS, before purification; and the purifying method is performed under native condition without using urea to avoid the problems of requiring lengthy processes to remove urea in purified protein solution or re-aggregation and precipitation of protein after removal of urea.
Claims9
A producing and purifying method of soluble recombinant COQ5 protein expressed in non-denaturing conditions, comprising steps of
- providing an E. coli host cell transformed with an expression vector containing DNA fragments of encoding sequence for soluble COQ5 protein;
- inducing the expressing of recombinant soluble COQ5 protein in the E. coli host cell;
- lysing the bacterial cells by a lysis buffer comprising lysozyme and DNase I;
- treating the cell lysate with low concentration of ionic detergent;
- centrifugation for removing insoluble proteins or debris and collecting supernatant containing the recombinant soluble COQ5 protein;
- and
- applying the supernatant to Ni-NTA beads under native condition (without using urea) to obtain purified recombinant soluble COQ5 protein.
9-14. (canceled)
Dependencies are inferred from the claim text.