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

Wang HD, Li FF, He C, Cui J, Song W, Li ML - J. Insect Sci. (2014)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Forestry Pests Biological Control, College of Forestry, Northwest A&F University, Yangling, Shaanxi, 712100, P. R China These authors contributed equally to this work wanghd2826@gmail.com.

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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.
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f04_01: 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.

Mentions: Though the cDNA fragments derived from different primers or contigs usually had <70% deduced amino acid identity, exceptions still existed. Some deduced amino acid sequences from different groups were found to be much closer to each other. For instance, according to Table 5 and Figure 4, the deduced amino acid identity of the corresponding regions of A1 (group I) and E7 (group II), H3 (group I) and E20 (group II) were both 97.5%, and that of H1 (group I) and E7 (group II), G5 (group II) and H3 (group I) were both 100%, which indicated that fragments H3, G5, and E20 might belong to the same P450 genes (CYP6BQ subfamily) or its allele, as well as F2, A20, and F6, which are supposed to be the parts of another P450 gene (CYP6BK subfamily) or its allele.


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

Wang HD, Li FF, He C, Cui J, Song W, Li ML - J. Insect Sci. (2014)

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.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4206240&req=5

f04_01: 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.
Mentions: Though the cDNA fragments derived from different primers or contigs usually had <70% deduced amino acid identity, exceptions still existed. Some deduced amino acid sequences from different groups were found to be much closer to each other. For instance, according to Table 5 and Figure 4, the deduced amino acid identity of the corresponding regions of A1 (group I) and E7 (group II), H3 (group I) and E20 (group II) were both 97.5%, and that of H1 (group I) and E7 (group II), G5 (group II) and H3 (group I) were both 100%, which indicated that fragments H3, G5, and E20 might belong to the same P450 genes (CYP6BQ subfamily) or its allele, as well as F2, A20, and F6, which are supposed to be the parts of another P450 gene (CYP6BK subfamily) or its allele.

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Forestry Pests Biological Control, College of Forestry, Northwest A&F University, Yangling, Shaanxi, 712100, P. R China These authors contributed equally to this work wanghd2826@gmail.com.

Show MeSH