Development of two novel CYP-antibodies and their use in a PCB exposure experiment with Mytilus edulis |
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| Authors: | H Jonsson K Viken Sandnes D Schiedek R Schneider B E Grsvik A Goksyr |
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| Institution: | a Department of Biological Oceanography, Baltic Sea Research Institute, Seestrasse 15, D-18119, Rostock, Germany;b Department of Molecular Biology, University of Bergen, HIB, P.O. Box 7800, N-5020, Bergen, Norway;c Biosense Laboratories AS, Thormøhlensgate 55, N-5008, Bergen, Norway |
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| Abstract: | In an attempt to learn more about the cytochrome P450 (CYP) system of mussels, we used protein databases and alignment software to extract highly conserved CYP sequences. From these alignments synthetic peptides were produced and used for rabbit immunisation, which yielded polyclonal antibodies against the CYP families 2 and 4. The antibodies were evaluated with Western Blot and ELISA assays, using digestive gland microsomal samples from the mussel Mytilus edulis. Western Blots revealed immunoreactions for both antibodies. The anti-CYP2 sequence rendered one major immunopositive protein of ≈49 kDa size, and weak signals for proteins of ≈41 and 56 kDa size. The anti-CYP4 sequence rendered two major bands of ≈56 and 59 kDa size, and also a weak immunoreaction with a protein of ≈43 kDa size. ELISA rendered only weak signals even with a 1:50 dilution of IgG-purified serum. A 10-day exposure to Aroclor 1254 did not appear to affect any of the immunopositive proteins, while total PCBs in soft bodies increased from 14–40 ng/g DW in controls to 373–638 ng/g DW in exposed mussels. |
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| Keywords: | Mussel Mytilus sp Mixed function oxidase Cytochrome P450 Chlorinated hydrocarbons Immunoreaction Western blot |
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