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Cloning and expression of Koala )Phascolarctos cinerus) liver cytochrome P450 reductase

Kong, S, Ngo, SN, McKinnon, RA and Stupans, L (2009). Cloning and expression of Koala )Phascolarctos cinerus) liver cytochrome P450 reductase. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology,150(1):1-9.

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Title Cloning and expression of Koala )Phascolarctos cinerus) liver cytochrome P450 reductase
Author Kong, S
Ngo, SN
McKinnon, RA
Stupans, L
Journal Name Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology
Publication Date 2009
Volume Number 150
Issue Number 1
ISSN 1532-0456   (check CDU catalogue open catalogue search in new window)
Start Page 1
End Page 9
Total Pages 9
Place of Publication US
Publisher Elsevier Inc.
HERDC Category C1 - Journal Article (DEST)
Abstract The 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 µM, 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.
DOI http://dx.doi.org/10.1016/j.cbpc.2009.01.010   (check subscription with CDU E-Gateway service for CDU Staff and Students  check subscription with CDU E-Gateway in new window)
 
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Created: Mon, 22 Mar 2010, 14:07:13 CST by Sarena Wegener