Journal of Tropical Oceanography >
Molecular cloning of cDNA core sequences of detoxification-related genes and liver constitutive expression in rabbitfish Siganus oramin
Received date: 2007-11-28
Revised date: 2008-04-09
Online published: 2009-12-12
Supported by
国家科技部“863”项目(2007AA09Z437), 国家自然科学基金项目(30670367), 广东省科技计划项目(2007B020701002;2005B20301005), 广东省自然科学基金项目(031886)和教育部留学回国人员科研启动基金项目
Partial cDNA sequences of hepatic cytochrome P-450 1A (CYP1A), glutathione S-transferase alpha (GSTA), glutathione S-transferase rho (GSTR), heat shock protein 70 (HSP70), Na+/K+-ATPase alpha 1(ATP1A1) and beta-actin (ACT) genes from rabbitfish (Siganus oramin) were obtained by PCR using degenerate primers. These sequences were 879 bp, 582 bp, 588 bp, 660 bp, 749 bp and 554 bp in length, respectively. Homology of the CYP1A, GSTA and GSTR amino acid sequence are high with bastard halibut (Paralichthys olivaceus), European plaice (Pleuronectes platessa), red sea bream (Pagrus major) and zebrafish CYP1A, GSTA and GSTR, and CYP1A and GSTA are low with human, rat, mouse, chicken and African clawed frog. The identities of HSP70, ATP1A and beta-actin amino acid sequences are highly conserved between fish, bird, amphibian and mammals. Using semi-quantitative PCR within the exponential phase, with beta-actin as an external control, the constitutive transcriptional expression of genes of phase I (CYP1A), phase II (GSTA and GSTR) xenobiotic metabolizing enzymes (XMEs), HSP70 and ATP1A1 were also investigated. The levels of CYP1A, GSTA and GSTR mRNA expression were higher, and the levels of HSP70 and ATP1A1 mRNA expression were lower. The constitutive expression pattern of these genes, show an accordant relationship with their functions, which provide data for our understanding to the detoxification mechanism of marine algae toxin in marine fish.
WANG Lin , LIANG Xu-fang , LIN Qun , Li Guang-zhao , Hu Yong-le . Molecular cloning of cDNA core sequences of detoxification-related genes and liver constitutive expression in rabbitfish Siganus oramin[J]. Journal of Tropical Oceanography, 2009 , 28(6) : 79 -87 . DOI: 10.11978/j.issn.1009-5470.2009.06.079
[1] White A W. Can. J. Fish. Aquat Sci, 1980, 37: 2262-2265.
[2] White A W. Paralytic shellfish toxins and finfish [A], In: Ragelis, E P (ed.) Seafood toxins [M]. ACS sympodium series 262. Amer Chem Soc, Washington D C, 1984, 171-180.
[3] Bricelj V M, LEE J H, CEMBELLA A D. Influence of dinoflagellate cell toxicity on uptake and loss of paralytic shellfish toxins in the northern quahog, Mercenaria mercenaria[J]. Mar Ecol Prog Ser, 1990, 74: 33-46.
[4] BRICELJ V M, LEE J H, ANDERSON D M. Uptake kinetics of paralytic shellfish toxins from the dinoflagellate Alexandrium fundyense in the mussel Mytilus edulis[J]. Mar Ecol Progr Ser, 1991, 63: 177-188.
[5] CHEN, C Y, CHOU, H N. Fate of paralytic shellfish poisoning toxins in purple clam Hiatula rostrata, in outdoor culture and laboratory culture[J]. Marine pollution Bulletin, 2002, 44: 733-738.
[6] SHUMWAY S E. Toxic algae, a threat to shellfish farming. World Aquacult, 1989, 20(3): 65-74.
[7] BEATTIE K A, RESSLER J, WIEGAND C, et al. Comparative effects and metabolism of two microcystins and nodularin in the brine shrimp Artemia salina [J]. Aquat Toxicol, 2003, 62: 219-226.
[8] GUBBINS M J, EDDY F B, GALLACHER S, STAGG R M. Paralytic shellfish poisoning toxins induce xenobiotic metabolizing enzymes in Atlantic salmon (Salmo salar)[J]. Marine Environmental Research, 2000, 50: 479-483.
[9] STAGG RM, GALLACHER S, BURGESS P. Harmful algae. In Reguera J B, Blanco M L, FernaÂndez & T Wyatt, Xunta de Galicia and Intergovernmental Oceanographic Commission of UNESCO[R]. UNESCO, Santiago de Compostela, 1998:607-608.?
[10]LIAO W Q, LIANG X F, WANG L, et al. Molecular cloning and characterization of alpha-class glutathione S-transferase gene from the liver of silver carp, bighead carp and other major Chinese freshwater fishes [J]. J Biochem Mol Toxicol, 2006, 20: 114-126.
[11]LIAO W Q, LIANG X F, WANG L, et al. Structural conservation and food habit-related liver expression of uncoupling protein 2 gene in five major Chinese freshwater fishes [J]. J Biochem Mol Biol, 2006, 39(4): 346-354.
[12]WANG L, LIANG X F, LIAO W Q, et al. Structural and functional characterization of microcystin detoxification-related liver genes in a phytoplanktivorous fish, Nile tilapia (Oreochromis niloticus) [J]. Comp Biochem Physiol C, 2006, 144: 216-227.
[13] FENG Q, BOONE A N, VIJAYAN M M. Copper impact on heat shock 70 expression and apoptosis in rainbow trout hepatocytes[J]. Comp Biochem Physiol C Toxicol Pharmacol, 2003, 135: 345-355.
[14] PAVLOV K V, SOKOLOV V S. Electrogenic Ion Transport by Na+/K+ -ATPase[J]. Membr Cell Biol, 2000, 13: 745-788.
[15] TANG J Y M, WONG C K C, AU D W T. The ichthyotoxic alga chattonella marina induces Na+, K+ -ATPase, and CFTR proteins expression in fish gill chloride cells in vivo[J]. Biochemical and Biophysical Research Communications, 2007, 353: 98-103.
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