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- Title
Identification of the proteins related to cytochrome P450 induced by fenvalerate in a Trichoplusia ni cell line.
- Authors
Xin-Kui Fang; Dan-Feng Huang; Zhao-Xia Wang; Chun-Ling Wan; Tao Sun; Wang-Jie Xu; Chun-Yan Liu; Pei Zhou; Zhong-Dong Qiao
- Abstract
Abstract  In order to reveal the metabolic reaction to the presence of fenvalerate mediated by P450 in insects, we used the trypan blue exclusion technique and 3-(4,5-dimethylthiazol)-2,5-diphenyltrazolium bromide (MTT) reduction assay to assess the vitality of Trichoplusia ni (Tn) cells treated with fenvalerate, and observed dose- and time-dependent changes in total cellular P450s. In addition, two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) were used to identify the proteins involved in the fenvalerate reaction process. Finally, the cDNA of P450 fragments was cloned and real-time RT-PCR was performed. Our data showed that at the 0â15 μmol/L challenge concentration of fenvalerate, at which the vitality of Tn cells was not affected (pâ>â0.05), there was a tendency toward a dose- and time-response of total cellular P450s, which peaked at the 9 h (pâpâP450 cDNA fragments (CYP4L4) was cloned and sequenced, and its expression in treated Tn cells increased significantly (pâ
- Subjects
ELECTROPHORESIS; POLYMERASE chain reaction; TRICHOPLUSIA; BROMIDES
- Publication
Cell Biology & Toxicology, 2007, Vol 23, Issue 6, p445
- ISSN
0742-2091
- Publication type
Article
- DOI
10.1007/s10565-007-9006-1