Generation and characterisation of monoclonal antibodies to Rainbow trout (Oncorhynchus mykiss) prion protein

被引:7
|
作者
Maddison, BC
Patel, S
James, RF
Conlon, HE
Oidtmann, B
Baier, M
Whitelam, GC
Gough, KC
机构
[1] Univ Leicester, Dept Biol, Biotechnol Grp, ADAS, Leicester LE1 7RH, Leics, England
[2] Univ Leicester, Dept Infect Immun & Inflammat, Leicester LE1 9HN, Leics, England
[3] Weymouth Lab, CEFAS, Weymouth DT4 8UB, Dorset, England
[4] Univ Munich, Inst Zool Fish Biol & Fisg Dis, D-80539 Munich, Germany
[5] Robert Koch Inst, D-13353 Berlin, Germany
关键词
prion protein; fish; Salmonidae; PrP; antibodies;
D O I
10.1016/j.jim.2005.09.001
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We report the production and characterisation of three monoclonal antibodies to the prion protein (PrP) of Rainbow trout (Oncorhynchus mykiss), a piscine protein with characteristic structural features common to mammalian prion protein. All of the antibodies were used to detect PrP in ELISA, Western blot and by immunohistochemistry. The antibodies showed specificity for certain genera of the Salmonidae, binding to PrP of Rainbow trout and Atlantic salmon (Salmo salar) but not to that from Arctic char (Salvelinus alpinus). Using the immunoreagents in Western blots, we demonstrated that O. mykiss PrP protein is a 64 kDa protein present in brain, spinal chord and optic nerve. PrP was not detected in a range of peripheral tissues: eye, heart, stomach, intestine, liver, kidney, spleen, muscle and skin. Furthermore, PrP could be detected in all brain regions studied: optic lobe, cerebrum/olfactory lobe, cerebellum, hypothalamus/pituitary and medulla oblongata and was widespread within these tissues as determined by immunohistochemistry. These immunoreagents provide specific tools to study the biology of Rainbow trout and Atlantic salmon PrP and any possible transmissible spongiform encephalopathy-like disease of these economically important fish species. (c) 2005 Elsevier B.V. All rights reserved.
引用
收藏
页码:202 / 210
页数:9
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