HETEROLOGOUS EXPRESSION OF RECOMBINANT MOUSE CYTOCHROME P450 2E1 IN ESCHERICHIA COLI

Abstract

Aim. The cloning of the mouse cytochrome P450 2E1 cDNA and obtaining of expression of cloned cDNA in E. coli. Methods. Polymerase chain reaction, cloning techniques, electrophoresis, Western blot. Results. For cloning of the mouse cytochrome P450 2E1 cDNA pCWOri+ expression vector and the strain E. coli DH5α were used. To obtain recombinant cytochrome P450 2E1 expression the cDNA gene sequence encoding the protein N-terminal fragment was modified, it was leading to deletion from 3 to 23 amino acid residues, nucleotide substitution in the second Ala codon (GCG to GCT) and increasing AT content in the next six codons by silent substitutions. Conclusions. For the first time by using the pCWOri+ expression vector the mouse cytochrome P450 2E1 cDNA (with this modification) was cloned and its expression was obtained in E. coli.

Keywords

• цитохром р450 2е1
• гетерологічна експресія в e. coli
• рекомбінантний білок сур2е1.