CLONING AND HIGH-LEVEL EXPRESSION OF CHICKEN APOCYTOCHROME-C GENE IN ESCHERICHIA-COLI

Chicken apocytochrome c gene with correct reading frame was easily clo ned through excision by polymerase chain reaction of the intron in the genomic clone of chicken cytochrome c gene, and was successfully over expressed in Escherichia coli by cloning into expression vector pET-3d under the control of T-7 promoter. Expressed protein can amount to as high as 40% of the total protein and mainly presents as inclusion bod y. Purification of chicken apocytochrome c from the inclusion body and characterization by SDS-PAGE, isoelectric focusing electrophoresis, a nd amino acid analysis showed that the purified apocytochrome c is ide ntical to that prepared from chicken heart cytochrome c by chemically depletion of heme.