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Expression in Escherichia coli of the major outer capsid protein of infectious pancreatic necrosis virus
Authors:Parimi Muralikrishna and Eric Wickstrom
Institution:

a Department of Chemistry, University of South Florida, Tampa, FL 33620, U.S.A.

b BDepartment of Biiochemistry and Molecular Biology, University of South Florida, Tampa, FL 33620, U.S.A.

c Department of Surgery, University of South Florida, Tampa,FL 33620, U.S.A.

Abstract:The gene for Escherichia coli translational initiation factor 3 (infC) has been inserted into an overexpression plasmid under the control of the bacteriophage T7 promoter. The infC plasmid was then used to transform a host with a chromosomal T7 RNA polymerase gene controlled by the lacUVS promoter. Induction of T7 RNA polymerase expression in the host cells resulted in a 200-fold overexpression of infC mRNA and a 100-fold overproduction of initiation factor 3. Rapid batch purification of biologically active IF3 yielded predominantly the long form of IF3, implying that the short form is an artifact of purification by traditional methods.
Keywords:Recombinant DNA  expression vector  IPTG induction  protein overproduction  IF3 purification
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