Summary:
The aim of this work is the construction of an expression system for in vitro synthesis of microsomal
monooxygenases P4501A1 and P4503A4, which catalyze oxidative transformations of most xenobiotics
in both animal and human organisms. cDNAs encoding both proteins were obtained following
the UBMTA protocol by the courtesy of holders, and amplified by established methods. Baculovirus
transfer vectors were used to clone these cDNAs. These vectors contain a strong polyhedrine
promoter surrounded by sequences homologous to that of baculovirus DNA, allowing the recombination
of the vector with the viral DNA, and hence the production of a protein. Established methods
and PCR were used to insert cDNA into the vectors, and the insertion was verified by the PCR
method with specific primers and using restriction endonucleases.
Key words:
P4501A1 – P4503A4 – baculovirus transfer vector
|