2019 ASHS Annual Conference

Cytochrome P450 2E1 (CYP2E1) enzyme encoded by cyp2e1 gene plays an important role in metabolism of heterogeneous organics in mammalian liver cells. The transgenic plant including recombinant mammalian CYP2E1 gene can metabolize various volatile organic compounds (VOCs). However, it is unclear the mechanism of expression control of CYP2E1 in plants. In this study, plasmid pCAMBIA2300 harboring the mammalian CYP2E1 was transformed into Ardisia pusilla mediated by Agrobacterium tumefaciens. The integration of the CYP2E1 and NPTII genes into the chromosome of the transgenic Ardisia plants were verified via genomic DNA polymerase chain reaction amplification. Southern blot analysis revealed the presence of multiple copies of CYP2E1 in the genome of transgenic plants. Expression of mRNA of CYP2E1 was confirmed by Northern blot analysis. Western blot analysis indicated 57-kDa of recombinant mammalian CYP2E1 with 2 putative N-glycosylation in transgenic Ardisia plants. The aniline 4-hydroxylase activity of mammalian CYP2E1 produced from transgenic Ardisia leaf was confirmed using aniline as a substrate.

Keywords: Cytochrome P450 2E1 (CYP2E1), Cauliflower mosaic virus 35S (CaMV35S) promoter, Agrobacterium tumefaciens, Transgenic Ardisia pusilla, Aniline hydroxylase activity