Abstract
CYP450 plays an important role in physiological metabolism. A CYP4G25 gene of P450 family was cloned from Antheraea pernyi using reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE-PCR). Sequence analysis revealed that this gene was 2112 bp long and has 97.5% identity with Antheraea yamamai CYP4G25. Semi-quantitative polymerase chain reaction (PCR) showed that the expression of A. perny i CYP4G25 was found in various tissues with no significant changes. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis demonstrated that a 63.6 KD recombinant protein was successfully expressed in Escherichia coli cells and its expression was not remarkably changed under induction by different isopropyl-β-D-thiogalactopyranoside (IPTG) concentration. Key words: Antheraea pernyi, CYP4G25, expression, cytochrome P450.
Highlights
Cytochrome P450s are involved in the metabolism of hormone, fatty acid, steroid, drug insecticide and phytotoxin (Mansuy et al, 1998; Hannemann et al, 2007; Isin et al, 2007; Hassanin et al, 2009)
A CYP4G25 gene of P450 family was cloned from Antheraea pernyi using reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE-PCR)
Nucleotide sequence analysis revealed that CYP4G25 cDNA contains a 111 bp 5’-untranslated sequence, a putative open reading frame (ORF) of 1674 bp, a 326 bp 3′untranslated region (3′UTR) and a putative polyadenylation signal
Summary
Cytochrome P450s are involved in the metabolism of hormone, fatty acid, steroid, drug insecticide and phytotoxin (Mansuy et al, 1998; Hannemann et al, 2007; Isin et al, 2007; Hassanin et al, 2009). Mammal P450s play a dominant role in clearing ingested compounds and controlling the systemic levels of chemical substrates (Ding and Kaminsky, 2003; Bowles et al, 2006), and plant P450s are essential for the biosynthesis of many compounds including phenylpropanoids, lipids, phytohormones and carotenoids (Schuler and Werk, 2003; Inoue, 2004). Some P450s are found to be related with the behavioral phenotypes, insecticide metabolism or inactivation of plant toxins (Dierick and Greenspan, 2006; Wang et al, 2008; Che-Mendoza et al, 2009; Ai et al, 2010). Lots of P450 genes have been isolated from more than 40 insect species (Chung et al, 2009), the exact roles of P450s in various animals remain to be explored
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