

FOLLOWUS
1.Hebei Key Laboratory of Preventive Veterinary Medicine, Hebei Normal University of Science and Technology, Qinhuangdao 066004, China
2.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
Jian LI, E-mail: lijian@ysfri.ac.cn
Received:08 October 2020,
Accepted:13 November 2020,
Online First:05 January 2021,
Published:2022-01
Scan QR Code
Hai REN, Jian LI, Ping LIU, et al. Cloning of
Hai REN, Jian LI, Ping LIU, et al. Cloning of
Metalloproteases represent a class of extracellular proteases found in
Vibrio anguillarum
that can generate toxic and pathogenic effects in turbot (
Scophthalmus maximus
). The toxicological effect partly results from oxidative damage due to the production of excessive reactive oxygen species (ROS). Catalase (CAT)
superoxide dismutase (SOD)
and glutathione peroxidase (GPx) are major antioxidant enzymes induced by various oxidative stresses and can scavenge peroxides generated in cells. To evaluate the effects of metalloprotease-induced ROS on the antioxidation defense mechanism of
S. maximus
head kidney cells
the cDNA of
CAT
gene (designated as
SmCAT
) was cloned and characterized.
SmCAT
comprises a 1 584-bp coding sequence that encodes a protein containing 527 amino acids with a poly(A) tail. Bioinformatics analysis revealed an active site signature sequence
a heme-ligand signature sequence
and three catalytic amino acid residues. The deduced
SmCAT
amino acid sequence shares a sequence similarity of 66.1%–92.4% with those of other species. Phylogenetic analysis revealed that SmCAT is classified with CAT of other fishes. Quantitative real-time PCR analysis showed that
SmCAT
was extensively expressed in all tested tissues
especially in blood. The expression of
SmCAT
SmMnSOD
and
SmGPx
were inhibited significantly in head kidney cells treated with metalloprotease from 12 to 24 h. In 6 to 24 h metalloprotease-treated groups compared to that of the untreated group
it was found that the production of ROS was markedly increased
and the mitochondrial membrane potential was decreased considerably. Hoechst 33342 staining revealed the presence of apoptotic bodies when the cells were incubated with 8.0 or 40.0 μg/mL metalloprotease for 12 and 24 h. Hence
the toxic effects of metalloprotease are associated with the down-regulation of antioxidant enzyme expression and increased ROS levels
which trigger the activation of apoptosis in the head kidney cells of turbot. Our findings provide a better understanding on the mechanism of metalloprotease-induced apoptosis in fish.
V Afonso , R Champy , D Mitrovic , P Collin , A Lomri . Reactive oxygen species and superoxide dismutases: role in joint diseases . Joint Bone Spine , 2007 . 74 ( 4 ): 324 - 329 . DOI: 10.1016/j.jbspin.2007.02.002 http://doi.org/10.1016/j.jbspin.2007.02.002 .
K W An , H S Shin , C Y Choi . Physiological responses and expression of metallothionein (MT) and superoxide dismutase (SOD) mRNAs in olive flounder, Paralichthys olivaceus exposed to benzo[a]pyrene . Comparative Biochemistry & Physiology Part B: Biochemistry & Molecular Biology , 2008 . 149 ( 3 ): 534 - 539 . DOI: 10.1016/j.cbpb.2007.12.004 http://doi.org/10.1016/j.cbpb.2007.12.004 .
A Avci , M Kaçmaz , I Durak . Peroxidation in muscle and liver tissues from fish in a contaminated river due to a petroleum refinery industry . Ecotoxicology and Environmental Safety , 2005 . 60 ( 1 ): 101 - 105 . DOI: 10.1016/j.ecoenv.2003.10.003 http://doi.org/10.1016/j.ecoenv.2003.10.003 .
P A Bain , K A Schuller . Molecular cloning of glutathione peroxidase cDNAs from Seriola lalandi and analysis of changes in expression in cultured fibroblast-like cells in response to tert -butyl hydroquinone . Aquaculture , 2012 . 324-325 182 - 193 . DOI: 10.1016/j.aquaculture.2011.10.027 http://doi.org/10.1016/j.aquaculture.2011.10.027 .
U Bandyopadhyay , D Das , R K Banerjee . Reactive oxygen species: oxidative damage and pathogenesis . Current Science , 1999 . 77 ( 5 ): 658 - 666 . .
J X Chen , B Liu , Z H Chi , Y Li , X H Zhang . Effect of culture conditions on the growth and extracellular protease expression of Vibrio anguillarum W1 . Marine Sciences , 2009 . 33 ( 1 ): 54 - 57 . .
Chen S Y. 2003. The Study of Major Virulent Factor of Vibrio anguillarum . The Institute of Oceanology, Chinese Academy of Sciences, Qingdao. (in Chinese with English abstract)
Chi Z H. 2006. Studies on Purification, Modification and Cytotoxicity of the Extracellular Metalloprotease of the Pathogenic Vibrio anguillarum . Ocean University of China, Qingdao. (in Chinese with English abstract)
M Cui , Y H Zhang , S S Liu , W D Xie , M Ji , H X Lou , X Li . 1-Oxoeudesm-11(13)-ene-12, 8α-lactone-induced apoptosis via ROS generation and mitochondria activation in MCF-7 cells . Archives of Pharmacal Research , 2011 . 34 ( 8 ): 1323 - 1329 . DOI: 10.1007/s12272-011-0812-x http://doi.org/10.1007/s12272-011-0812-x .
Cui Y T. 2014. Effects of Some Important Exogenous Factors on Cell Viability, Oxidative Damage, Apoptosis and Heat Stock Protein Expression in Hepatic Cells of Grass Crap ( Ctenopharyngodon idellaus ). Nanjing Agricultural University, Nanjing. (in Chinese with English abstract)
S M Denkin , D R Nelson . Regulation of Vibrio anguillarum empA metalloprotease expression and its role in virulence . Applied and Environmental Microbiology , 2004 . 70 ( 7 ): 4193 - 4204 . DOI: 10.1128/AEM.70.7.4193-4204.2004 http://doi.org/10.1128/AEM.70.7.4193-4204.2004 .
S Desagher , J C Martinou . Mitochondrial as the central control point of apoptosis . Trends in Cell Biology , 2000 . 10 ( 9 ): 369 - 377 . DOI: 10.1016/s0962-8924(00)01803-1 http://doi.org/10.1016/s0962-8924(00)01803-1 .
P M Ekanayake , M De Zoysa , H S Kang , Q Wan , Y Jee , Y H Lee , S J Kim , J Lee . Cloning, characterization and tissue expression of disk abalone ( Haliotis discus discus ) catalase . Fish & Shellfish Immunology , 2008 . 24 ( 3 ): 267 - 278 . DOI: 10.1016/j.fsi.2007.11.007 http://doi.org/10.1016/j.fsi.2007.11.007 .
D A S Elvitigala , H K A Premachandra , I Whang , T T Priyathilaka , E Kim , B S Lim , H B Jung , S Y Yeo , H C Park , J Lee . Marine teleost ortholog of catalase from rock bream ( Oplegnathus fasciatus ): molecular perspectives from genomic organization to enzymatic behavior with respect to its potent antioxidant properties . Fish & Shellfish Immunology , 2013 . 35 ( 4 ): 1086 - 1096 . DOI: 10.1016/j.fsi.2013.07.013 http://doi.org/10.1016/j.fsi.2013.07.013 .
D A S Elvitigala , T T Priyathilaka , I Whang , B H Nam , J Lee . A teleostan homolog of catalase from black rockfish ( Sebastes schlegelii ): insights into functional roles in host antioxidant defense and expressional responses to septic conditions . Fish & Shellfish Immunology , 2015 . 44 ( 1 ): 321 - 331 . DOI: 10.1016/j.fsi.2015.02.020 http://doi.org/10.1016/j.fsi.2015.02.020 .
M X Ge , X M Jin , H Fang , Y Y Zhang , C Z Chen . Detection and sequence analysis of virulence genes of Vibrio anguillarum isolated from turbot ( Scophthalmus maximus ) . Chinese Journal of Veterinary Science , 2014 . 34 ( 8 ): 1267 - 1271, 1277 . .
G S Gerhard , E J Kauffman , M A Grundy . Molecular cloning and sequence analysis of the Danio rerio catalase gene . Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology , 2000 . 127 ( 4 ): 447 - 457 . DOI: 10.1016/s0305-0491(00)00285-6 http://doi.org/10.1016/s0305-0491(00)00285-6 .
S Gül , E Belge-Kurutas , E Yıldız , A Şahan , F Doran . Pollution correlated modifications of liver antioxidant systems and histopathology of fish (Cyprinidae) living in Seyhan Dam Lake, Turkey . Environment International , 2004 . 30 ( 5 ): 605 - 609 . DOI: 10.1016/S0160-4120(03)00059-x http://doi.org/10.1016/S0160-4120(03)00059-x .
D D Guo , H S Bi , B Liu , Q X Wu , D G Wang , Y Cui . Reactive oxygen species-induced cytotoxic effects of zinc oxide nanoparticles in rat retinal ganglion cells . Toxicology in Vitro , 2013 . 27 ( 2 ): 731 - 738 . DOI: 10.1016/j.tiv.2012.12.001 http://doi.org/10.1016/j.tiv.2012.12.001 .
Y F Han , Z L Mo , P Xiao , B Hao , J Li , G P Yang . Characterization of EmpA protease in Vibrio anguillarum M3 . Journal of Ocean University of China , 2011 . 10 ( 4 ): 379 - 384 . DOI: 10.1007/s11802-011-1781-x http://doi.org/10.1007/s11802-011-1781-x .
D E Heck , M Shakarjian , H D Kim , J D Laskin , A M Vetrano . Mechanisms of oxidant generation by catalase . Annals of the New York Academy of Sciences , 2010 . 1203 ( 1 ): 120 - 125 . DOI: 10.1111/j.1749-6632.2010.05603.x http://doi.org/10.1111/j.1749-6632.2010.05603.x .
H Inamura , T Nakai , K Muroga . An extracellular protease produced by Vibrio anguillarum . Bulletin of the Japanese Society of Scientific Fisheries , 1985 . 51 ( 12 ): 1915 - 1920 . DOI: 10.2331/suisan.51.1915 http://doi.org/10.2331/suisan.51.1915 .
P G Jo , Y K Choi , C Y Choi . Cloning and mRNA expression of antioxidant enzymes in the Pacific oyster, Crassostrea gigas in response to cadmium exposure . Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology , 2008 . 147 ( 4 ): 460 - 469 . DOI: 10.1016/j.cbpc.2008.02.001 http://doi.org/10.1016/j.cbpc.2008.02.001 .
H Kamata , S I Honda , S Maeda , L F Chang , H Hirata , M Karin . Reactive oxygen species promote TNFα-induced death and sustained JNK activation by inhibiting MAP kinase phosphatases . Cell , 2005 . 120 ( 5 ): 649 - 661 . DOI: 10.1016/j.cell.2004.12.041 http://doi.org/10.1016/j.cell.2004.12.041 .
C F Ken , C T Lin , J L Wu , J F Shaw . Cloning and expression of a cDNA coding for catalase from zebrafish ( Danio rerio ) . Journal of Agricultural and Food Chemistry , 2000 . 48 ( 6 ): 2092 - 2096 . DOI: 10.1021/jf990838 http://doi.org/10.1021/jf990838 .
J Kim , N C N Perera , G I Godahewa , T T Priyathilaka , J Lee . Characterization of a catalase from red-lip mullet ( Liza haematocheila ): demonstration of antioxidative activity and mRNA upregulation in response to immunostimulants . Gene , 2019 . 712 143945 DOI: 10.1016/j.gene.2019.143945 http://doi.org/10.1016/j.gene.2019.143945 .
K J Kim , K Y Yoon , H D Hong , B Y Lee . Schisandra chinensis prevents hepatic lipid peroxidation and oxidative stress in rats subjected to heat environmental stress . Phytotherapy Research , 2012 . 26 ( 11 ): 1674 - 1680 . DOI: 10.1002/ptr.4640 http://doi.org/10.1002/ptr.4640 .
M G Klotz , P C Loewen . The molecular evolution of catalatic hydroperoxidases: evidence for multiple lateral transfer of genes between prokaryota and from bacteria into eukaryota . Molecular Biology and Evolution , 2003 . 20 ( 7 ): 1098 - 1112 . DOI: 10.1093/molbev/msg129 http://doi.org/10.1093/molbev/msg129 .
H Z Lee , W Z Liu , W T Hsieh , F Y Tang , J G Chung , H W C Leung . Oxidative stress involvement in Physalis angulata -induced apoptosis in human oral cancer cells . Food and Chemical Toxicology , 2009 . 47 ( 3 ): 561 - 570 . DOI: 10.1016/j.fct.2008.12.013 http://doi.org/10.1016/j.fct.2008.12.013 .
S Lee , K Söderhäll . Early events in crustacean innate immunity . Fish & Shellfish Immunology , 2002 . 12 ( 5 ): 421 - 437 . DOI: 10.1006/fsim.2002.0420 http://doi.org/10.1006/fsim.2002.0420 .
C H Li , D J Ni , L S Song , J M Zhao , H Zhang , L Li . Molecular cloning and characterization of a catalase gene from Zhikong scallop Chlamys farreri . Fish & Shellfish Immunology , 2008 . 24 ( 1 ): 26 - 34 . DOI: 10.1016/j.fsi.2007.06.010 http://doi.org/10.1016/j.fsi.2007.06.010 .
K J Livak , T D Schmittgen . Analysis of relative gene expression data using real-time quantitative PCR and the 2 -?? C t method . Methods , 2001 . 25 ( 4 ): 402 - 408 . DOI: 10.1006/meth.2001.1262 http://doi.org/10.1006/meth.2001.1262 .
S Lortz , M Tiedge , T Nachtwey , A E Karlsen , J Nerup , S Lenzen . Protection of insulin-producing RINm5F cells against cytokine-mediated toxicity through overexpression of antioxidant enzymes . Diabetes , 2000 . 49 ( 7 ): 1123 - 1130 . DOI: 10.2337/diabetes.49.7.1123 http://doi.org/10.2337/diabetes.49.7.1123 .
J L Martindale , N J Holbrook . Cellular response to oxidative stress: signaling for suicide and survival . Journal of Cellular Physiology , 2002 . 192 ( 1 ): 1 - 15 . DOI: 10.1002/jcp.10119 http://doi.org/10.1002/jcp.10119 .
Z L Mo , S Y Chen , P J Zhang . Properties of proteolytic toxin of Vibrio anguillarum from diseased flounder . Chinese Journal of Oceanology and Limnology , 2002 . 20 ( 4 ): 316 - 322 . DOI: 10.1007/BF02847921 http://doi.org/10.1007/BF02847921 .
J Nordberg , E S J Arnér . Reactive oxygen species, antioxidants, and the mammalian thioredoxin system . Free Radical Biology and Medicine , 2001 . 31 ( 11 ): 1 287 - 1 312 . DOI: 10.1016/S0891-5849(01)00724-9 http://doi.org/10.1016/S0891-5849(01)00724-9 .
C D Nwani , N S Nagpure , R Kumar , B Kushwaha , W S Lakra . DNA damage and oxidative stress modulatory effects of glyphosate-based herbicide in freshwater fish, Channa punctatus . Environmental Toxicology and Pharmacology , 2013 . 36 ( 2 ): 539 - 547 . DOI: 10.1016/j.etap.2013.06.001 http://doi.org/10.1016/j.etap.2013.06.001 .
M Planas , M Pérez-Lorenzo , J A Vázquez , J Pintado . A model for experimental infections with vibrio ( listonella ) anguillarum in first feeding turbot ( Scophthalmus maximus L.) larvae under hatchery conditions . Aquaculture , 2005 . 250 ( 1-2 ): 232 - 243 . DOI: 10.1016/j.aquaculture.2005.04.050 http://doi.org/10.1016/j.aquaculture.2005.04.050 .
Z T Qi , Q H Zhang , Z S Wang , W H Zhao , S N Chen , Q Gao . Molecular cloning, expression analysis and functional characterization of interleukin-22 in So-iny mullet, Liza haematocheila . Molecular Immunology , 2015 . 63 ( 2 ): 245 - 252 . DOI: 10.1016/j.molimm.2014.07.006 http://doi.org/10.1016/j.molimm.2014.07.006 .
Z Y Qian , T Liu , Q Liu , S L He , Y J Liu , F J Hou , X Z Wang , X Mi , C Cai , X L Liu . p53 is involved in shrimp survival via its regulation roles on MnSOD and GPx in response to acute environmental stresses . Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology , 2014 . 159 38 - 51 . DOI: 10.1016/j.cbpc.2013.09.009 http://doi.org/10.1016/j.cbpc.2013.09.009 .
L Qin , Y G Wang , B L Yan . Advances in research on diseases of turbot Scophthalmus maximus affected by microorganisms . Fisheries Science , 2008 . 27 ( 11 ): 598 - 602 . DOI: 10.3969/j.issn.1003-1111.2008.11.012 http://doi.org/10.3969/j.issn.1003-1111.2008.11.012 .
H Ren , J Li , J T Li , Y Ying , H X Ge , D L Li , T J Yu . Cloning of catalase and expression patterns of catalase and selenium-dependent glutathione peroxidase from Exopalaemon carinicauda in response to low salinity stress . Acta Oceanologica Sinica , 2015 . 34 ( 8 ): 52 - 61 . DOI: 10.1007/s13131-015-0640-9 http://doi.org/10.1007/s13131-015-0640-9 .
J L Rock , D R Nelson . Identification and characterization of a hemolysin gene cluster in Vibrio anguillarum . Infection and Immunity , 2006 . 74 ( 5 ): 2777 - 2786 . DOI: 10.1128/IAI.74.5.2777-2786.2006 http://doi.org/10.1128/IAI.74.5.2777-2786.2006 .
S Sellaththurai , T T Priyathilaka , J Lee . Molecular cloning, characterization, and expression level analysis of a marine teleost homolog of catalase from big belly seahorse ( Hippocampus abdominalis ) . Fish & Shellfish Immunology , 2019 . 89 647 - 659 . DOI: 10.1016/j.fsi.2019.03.064 http://doi.org/10.1016/j.fsi.2019.03.064 .
V Selvaraj , M Y Armistead , M Cohenford , E Murray . Arsenic trioxide (As 2 O 3 ) induces apoptosis and necrosis mediated cell death through mitochondrial membrane potential damage and elevated production of reactive oxygen species in PLHC-1 fish cell line . Chemosphere , 2013 . 90 ( 3 ): 1201 - 1209 . DOI: 10.1016/j.chemosphere.2012.09.039 http://doi.org/10.1016/j.chemosphere.2012.09.039 .
Speit G, Rothfuss A. 2012. The comet assay: a sensitive genotoxicity test for the detection of DNA damage and repair. In : DNA Repair Protocols. Methods in Molecular Biology. Humana Press, Totowa, NJ. 920 : 79-90, https://doi.org/10.1007/978-1-61779-998-3_6 https://doi.org/10.1007/978-1-61779-998-3_6 .
A R D Stebbing . Hormesis-the stimulation of growth by low levels of inhibitors . Science of the Total Environment , 1982 . 22 ( 3 ): 213 - 234 . DOI: 10.1016/0048-9697(82)90066-3 http://doi.org/10.1016/0048-9697(82)90066-3 .
S M Sun , X P Ge , J Zhu , F J Xuan , X J Jiang . Identification and mRNA expression of antioxidant enzyme genes associated with the oxidative stress response in the Wuchang bream ( Megalobrama amblycephala Yih) in response to acute nitrite exposure . Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology , 2014 . 159 69 - 77 . DOI: 10.1016/j.cbpc.2013.09.005 http://doi.org/10.1016/j.cbpc.2013.09.005 .
K Tamura , J Dudley , M Nei , S Kumar . MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0 . Molecular Biology and Evolution , 2007 . 24 ( 8 ): 1596 - 1599 . DOI: 10.1093/molbev/msm092 http://doi.org/10.1093/molbev/msm092 .
O L Tavares-Sánchez , G A Gómez-Anduro , X Felipe-Ortega , M A Islas-Osuna , R R Sotelo-Mundo , C Barillas-Mury , G Yepiz-Plascencia . Catalase from the white shrimp Penaeus ( Litopenaeus ) vannamei : molecular cloning and protein detection . Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology , 2004 . 138 ( 4 ): 331 - 337 . DOI: 10.1016/j.cbpc.2004.03.005 http://doi.org/10.1016/j.cbpc.2004.03.005 .
A E Toranzo , B Magariños , J L Romalde . A review of the main bacterial fish diseases in mariculture systems . Aquaculture , 2005 . 246 ( 1-4 ): 37 - 61 . DOI: 10.1016/j.aquaculture.2005.01.002 http://doi.org/10.1016/j.aquaculture.2005.01.002 .
M Varina , S M Denkin , A M Staroscik , D R Nelson . Identification and characterization of Epp, the secreted processing protease for the Vibrio anguillarum EmpA metalloprotease . Journal of Bacteriology , 2008 . 190 ( 20 ): 6589 - 6597 . DOI: 10.1128/jb.00535-08 http://doi.org/10.1128/jb.00535-08 .
N Wang , X L Wang , Z X Sha , Y S Tian , S L Chen . Development and characterization of a new marine fish cell line from turbot ( Scophthalmus maximus ) . Fish Physiology and Biochemistry , 2010 . 36 ( 4 ): 1227 - 1234 . DOI: 10.1007/s10695-010-9402-y http://doi.org/10.1007/s10695-010-9402-y .
Y X Wei , J C Wang , D L Cheng , Y L Xu . Purification and property of extracellular proteases from Vibrio anguillarum . Chinese Journal of Applied and Environmental Biology , 2002 . 8 ( 4 ): 414 - 418 . DOI: 10.3321/j.issn:1006-687X.2002.04.017 http://doi.org/10.3321/j.issn:1006-687X.2002.04.017 .
S Woo , S Yum , D W Kim , H S Park . Transcripts level responses in a marine medaka ( Oryzias javanicus ) exposed to organophosphorus pesticide . Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology , 2009 . 149 ( 3 ): 427 - 432 . DOI: 10.1016/j.cbpc.2008.10.100 http://doi.org/10.1016/j.cbpc.2008.10.100 .
K Yamamoto , Y Banno , H Fujii , F Miake , N Kashige , Y Aso . Catalase from the silkworm, Bombyx mori : gene sequence, distribution, and overexpression . Insect Biochemistry and Molecular Biology , 2005 . 35 ( 4 ): 277 - 283 . DOI: 10.1016/j.ibmb.2005.01.001 http://doi.org/10.1016/j.ibmb.2005.01.001 .
L F Yan , X An , M M Bao , J Y He , C W Wu , H H Liu . Expression of catalase in Larimichthys crocea after infection of Vibrio anguillarum . Journal of Fisheries of China , 2017 . 41 ( 5 ): 641 - 648 . DOI: 10.11964/jfc.20160110231 http://doi.org/10.11964/jfc.20160110231 .
H Yang , J X Chen , G P Yang , X H Zhang , Y Li , M Wang . Characterization and pathogenicity of the zinc metalloprotease EmpA of Vibrio anguillarum expressed in Escherichia coli . Current Microbiology , 2007 . 54 ( 3 ): 244 - 248 . DOI: 10.1007/s00284-006-0495-6 http://doi.org/10.1007/s00284-006-0495-6 .
H B Yu , W Deng , D D Zhang , Y Gao , Z Yang , X C Shi , J Sun , J S Zhou , H Ji . Antioxidant defenses of onychostoma macrolepis in response to thermal stress: insight from mRNA expression and activity of superoxide dismutase and catalase . Fish & Shellfish Immunology , 2017 . 66 50 - 61 . DOI: 10.1016/j.fsi.2017.04.027 http://doi.org/10.1016/j.fsi.2017.04.027 .
B Yun , Q H Ai , K S Mai , W Xu , G S Qi , Y W Luo . Synergistic effects of dietary cholesterol and taurine on growth performance and cholesterol metabolism in juvenile turbot ( Scophthalmus maximus L.) fed high plant protein diets . Aquaculture , 2012 . 324 ( -325 ): 85 - 91 . DOI: 10.1016/j.aquaculture.2011.10.012 http://doi.org/10.1016/j.aquaculture.2011.10.012 .
H C Zhang , K X Ma , Y J Yang , C Y Shi , G Y Chen , D Z Liu . Molecular cloning, characterization, expression and enzyme activity of catalase from planarian Dugesia japonica in response to environmental pollutants . Ecotoxicology and Environmental Safety , 2018 . 165 88 - 95 . DOI: 10.1016/j.ecoenv.2018.08.083 http://doi.org/10.1016/j.ecoenv.2018.08.083 .
0
Views
0
Downloads
1
CSCD
Publicity Resources
Related Articles
Related Author
Related Institution
京公网安备11010802024621