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. 2014 Feb 26:14:28.
doi: 10.1093/jis/14.1.28.

Molecular cloning and sequence analysis of novel cytochrome P450 cDNA fragments from Dastarcus helophoroides

Hai-Dong Wang  1 , Fei-Fei Li  2 , Cai He  3 , Jun Cui  4 , Wang Song  5 , Meng-Lou Li  6
Affiliations

Molecular cloning and sequence analysis of novel cytochrome P450 cDNA fragments from Dastarcus helophoroides

Hai-Dong Wang et al. J Insect Sci. .

Abstract

The predatory beetle Dastarcus helophoroides (Fairmaire) (Coleoptera: Bothrideridae) is a natural enemy of many longhorned beetles and is mainly distributed in both China and Japan. To date, no research on D. helophoroides P450 enzymes has been reported. In our study, for the better understanding of P450 enzymes in D. helophoroides, 100 novel cDNA fragments encoding cytochrome P450 were amplified from the total RNA of adult D. helophoroides abdomens using five pairs of degenerate primers designed according to the conserved amino acid sequences of the CYP6 family genes in insects through RT-PCR. The obtained nucleotide sequences were 250 bp, 270 bp, and 420 bp in length depending on different primers. Ninety-six fragments were determined to represent CYP6 genes, mainly from CYP6BK, CYP6BQ, and CYP6BR subfamilies, and four fragments were determined to represent CYP9 genes. Twenty-two fragments, submitted to GenBank, were selected for further homologous analysis, which revealed that some fragments of different sizes might be parts of the same P450 gene.

Keywords: CYP6 fragments; homologous analysis.

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Figures

Figure 1.
Figure 1.
PCR products by different primers. Products A, E, F, G, and H were derived by primers dpA, dpE, dpF, dpG, and dpH, respectively. Product lengths (bp) from left to right are: 250, 420, 270, 420, 270. M = marker. High quality figures are available online.
Figure 2.
Figure 2.
Deduced amino acid alignments of products A, F and H. High quality figures are available online.
Figure 3.
Figure 3.
Deduced amino acid alignments of products E and G. High quality figures are available online.
Figure 4.
Figure 4.
Phylogenetic relationships based on the amino acid sequence alignments of 22 novel sequences and P450 genes from CYP6 and CYP9 families. Letter designation: Phc: Pediculus humanus corporis ; Tc: Tribolium castaneum . High quality figures are available online.
See this image and copyright information in PMC

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