Issue:ISSN 2095-1353
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2012年49 No.4
Molecular cloning and mRNA expression analysis of cryptochrome
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Key Words:Agrotis ypsilon,cryptochrome,mRNA expression
Abstract: Cryptochrome ( cry) are blue and UV light photoreceptors and also act as an integral component of circadian clocks. In this study,two black cutworm ( Agrotis ypsilon ) cryptochrome genes,named Aycry1 ( GeneBank No.JQ616846) and Aycry2 ( GeneBank No. JQ616847 ),were cloned and characterized by RT-PCR and SMART RACE strategy. Their AUG-initiated ORFs ( open reading frames ) were 1 587 bp and 2 439 bp in length and encoded polypeptides were comprised of 528 and 812 amino acids respectively. Conserved domain analysis suggests that both Aycry1 and Aycry2 contain two basic conserved domains,the PHR region,which provides FAD binding sites,and the Cterminal conserved domain. Phylogenetic analysis indicates that insect CRYs are divided into two subgroups,CRY1 and CRY2,and AyCRY1 and AyCRY2 cluster with Lepidopteran CRY1 and CRY2 genes,respectively. Diel fluctuation of both Aycry1 and Aycry2 mRNA levels was determined by quantitative PCR analysis. The mRNA levels of both Aycry1 and Aycry2 were higher during the day than at night peaking at ZT7.