Kong, S.Ngo, S.McKinnon, R.Stupans, I.2011-10-072011-10-072009Comparative Biochemistry and Physiology Part - C: Toxicology and Pharmacology, 2009; 150(1):1-91532-04561878-1659http://hdl.handle.net/2440/66529The cloning, expression and characterization of hepatic NADPH-cytochrome P450 reductase (CPR) from koala (Phascolarctos cinereus) is described. Two 2059 bp koala liver CPR cDNAs, designated CPR1 and CPR2, were cloned by reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends. The koala CPR cDNAs encode proteins of 678 amino acids and share 85% amino acid sequence identity to human CPR. Transfection of the koala CPR cDNAs into Cos-7 cells resulted in the expression of proteins, which were recognized by a goat-antihuman CPR antibody. The koala CPR1 and 2 cDNA-expressed enzymes catalysed cytochrome c reductase at the rates of 4.9 +/- 0.5 and 2.6 +/- 0.4 nmol/min/mg protein (mean +/- SD, n = 3), respectively which were comparable to that of rat CPR cDNA-expressed enzyme. The apparent Km value for CPR activity in koala liver microsomes was 11.61 +/- 6.01 microM, which is consistent with that reported for rat CPR enzyme. Northern analysis detected a CPR mRNA band of approximately 2.6 kb. Southern analysis suggested a single PCR gene across species. The present study provides primary molecular data regarding koala CPR1 and CPR2 genes in this unique marsupial species.enCopyright © 2009 Elsevier Inc. All rights reserved.NADPH-cytochrome P450 reductaseRT-PCRCos-7 cellsSequence identityProteinmRNA levelJournal article002010594310.1016/j.cbpc.2009.01.0102-s2.0-6734920033830939Ngo, S. [0000-0002-9050-6894]