REGULAR ARTICLEA Universal Approach to the Expression of Human and Rabbit Cytochrome P450s of the 2C Subfamily inEscherichia coli
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Biophysical characterization of Aptenodytes forsteri cytochrome P450 aromatase
2018, Journal of Inorganic BiochemistryHeterologous expression and characterization of plant Taxadiene-5α-Hydroxylase (CYP725A4) in Escherichia coli
2017, Protein Expression and PurificationCitation Excerpt :The Kyte-Dolittle plot of wild-type CYP725A4 reveals that the N-terminus of the protein is highly hydrophobic (Supplemental Fig. S2). Truncating and modifying the hydrophobic N-terminal regions of CYPs has been shown to increase expression without affecting protein activity [37–40]. Previously, several modified versions of CYP725A4 were created [9].
Structural and kinetic basis of steroid 17α,20-lyase activity in Teleost fish cytochrome P450 17A1 and its absence in cytochrome P450 17A2
2015, Journal of Biological ChemistryCitation Excerpt :RNA was extracted from fresh zebrafish ovaries, and the cDNA was amplified by RT-PCR (Qiagen One Step RT-PCR kit). The region encoding the N-terminal transmembrane helix (residues 1–25) was replaced by DNA coding for MAKKTSSKGK (P450 2C3 N-terminal region (32)); the 3′ end was extended by 18 nucleotides encoding six histidines, and the modified cDNA was inserted into a pET17B vector (EMD Millipore). Zebrafish P450 17A1 and 17A2 expression and purification for enzymatic experiments were as follows: the P450 plasmids and a pGro12 (ES/EL) expression vector (33) were transformed into E. coli BL21-Gold (DE3) competent cells.
Pharmacogenomics of Clopidogrel
2014, Handbook of Pharmacogenomics and Stratified Medicine