Molecular cloning and sequence analysis of novel cytochrome P450 cDNA fragments from Dastarcus helophoroides.
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.
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.
Hai-Dong
HD
Wang
College of Science, Northwest A&F University, Yangling, Shaanxi, 712100, P. R China These authors contributed equally to this work feifeinewday@gmail.com.
Fei-Fei
FF
Li
Wuwei Academy of Forestry Sciences, Wuwei, Gansu 733000, P.R. China brittany0825@hotmail.com.
Cai
C
He
Laboratory of Forestry Pests Biological Control, College of Forestry, Northwest A&F University, Yangling, Shaanxi, 712100, P. R China cuijunclaire@hotmail.com.
Jun
J
Cui
Laboratory of Forestry Pests Biological Control, College of Forestry, Northwest A&F University, Yangling, Shaanxi, 712100, P. R China csongwang0516@yahoo.cn.
Wang
W
Song
Laboratory of Forestry Pests Biological Control, College of Forestry, Northwest A&F University, Yangling, Shaanxi, 712100, P. R China limenglou@hotmail.com.
Meng-Lou
ML
Li
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D001517:Coleoptera / Q000235:genetics* / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment
D018076:DNA, Complementary; D019476:Insect Proteins; D003577:Cytochrome P-450 Enzyme System
CYP6 fragments; homologous analysis