Isolation and Nucleotide Sequence Characterization of Novel Cytochrome P450 Hydroxylase Genes from Rare Actinomycetes, Sebekia benihana
Park, N.S.;Kim, E.S.(Inha University, Incheon, Republic of Korea)E-mail:[email protected] | Park, H.J.(SongWon Envichem, Co., Seoul, Republic of Korea) | Han, K.B.(Hanson Biotech Co., Ltd. 201 IACRI, Han Nam University, Daejeon, Republic of Korea) | Kim, S.N.(LG Household and HealthCare, RnD Park, Daejeon, Republic of Korea)
A degenerate set of PCR primers based on two conserved regions (heme binding region and oxygen ligand pocket) were designed and successfully applied to amplify DNA fragments of cytochrome P450 hydroxylase (CYP) genes from a rare actinomycetes, S. benihana. The PCR amplified products were employed as a DNA probe to clone the entire CYP genes from S. benihana genomic library. Five different CYP-positive cosmids were isolated by colony hybridization as well as PCR confirmation. The complete nucleotide sequencing of five different CYP genes revealed that each unique CYP showed a significant amino acid homology to previously-known CYP genes involved in streptomycetes secondary metabolism.
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