Identification and expression analysis of CYP4G25 gene from the Chinese oak silkworm (Antheraea pernyi)

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. pernyi 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.