Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
J Zhejiang Univ Sci B
2016 Feb 01;172:110-26. doi: 10.1631/jzus.B1500008.
Show Gene links
Show Anatomy links
Developing antibodies from cholinesterase derived from prokaryotic expression and testing their feasibility for detecting immunogen content in Daphnia magna.
???displayArticle.abstract???
To yield cholinesterase (ChE) from prokaryotic expression, the ChE gene that belongs to Daphnia magna was amplified by reverse transcription-polymerase chain reaction (RT-PCR) using forward primer 5'-CCCYGGNGCSAT GATGTG-3' and reverse primer 5'-GYAAGTTRGCCCAATATCT-3'. To express the gene, one sequence of the amplified DNA, which was able to encode a putative protein containing two conserved carboxylesterase domains, was connected to the prokaryotic expression vector PET-29a(+). The recombinant vector was transformed into Escherichia coil BL21 (DE3). Protein expression was induced by isopropy-D-thiogalactoside. The expressed ChE was used as an immunogen to immunize BALB/c mice. The obtained antibodies were tested for their specificity towards crude enzymes from species such as Alona milleri, Macrobrachium nipponense, Bombyx mori, Chironomus kiiensis, Apis mellifera, Eisenia foetida, Brachydanio rerio, and Xenopus laevis. Results indicated that the antibodies had specificity suitable for detecting ChE in Daphnia magna. A type of indirect and non-competitive enzyme-linked immunosorbent assay (IN-ELISA) was used to test the immunoreactive content of ChE (ChE-IR) in Daphina magna. The detection limit of the IN-ELISA was found to be 14.5 ng/ml at an antiserum dilution of 1:22 000. Results from tests on Daphnia magna exposed to sublethal concentrations of triazophos indicated a maximal induction of 57.2% in terms of ChE-IR on the second day after the animals were exposed to a concentration of 2.10 μg/L triazophos. Testing on animals acclimatized to a temperature of 16 °C indicated that ChE-IR was induced by 16.9% compared with the ChE-IR content detected at 21 °C, and the rate of induction was 25.6% at 10 °C. The IN-ELISA was also used to test the stability of ChE-IR in collected samples. Repeated freezing and thawing had no influence on the outcome of the test. All these results suggest that the polyclonal antibodies developed against the recombinant ChE are as efficient as those developed against the native ChE in detecting ChE content in Daphnia magna.
Abdel-Halim,
Organophosphorus pollutants (OPP) in aquatic environment at Damietta Governorate, Egypt: implications for monitoring and biomarker responses.
2006, Pubmed
Abdel-Halim,
Organophosphorus pollutants (OPP) in aquatic environment at Damietta Governorate, Egypt: implications for monitoring and biomarker responses.
2006,
Pubmed Anthony,
Cloning, sequencing and functional expression of an acetylcholinesterase gene from the yellow fever mosquito Aedes aegypti.
1995,
Pubmed Barata,
Biochemical factors contributing to response variation among resistant and sensitive clones of Daphnia magna Ssraus exposed to ethyl parathion.
2001,
Pubmed Botté,
Temperature: a prolonged confounding factor on cholinesterase activity in the tropical reef fish Acanthochromis polyacanthus.
2013,
Pubmed Bradford,
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
1976,
Pubmed Cajaraville,
The use of biomarkers to assess the impact of pollution in coastal environments of the Iberian Peninsula: a practical approach.
2000,
Pubmed Carvalho,
Use of atropine-treated Daphnia magna survival for detection of environmental contamination by acetylcholinesterase inhibitors.
2003,
Pubmed Coelho,
Assessing lethal and sub-lethal effects of trichlorfon on different trophic levels.
2011,
Pubmed Damásio,
Biochemical mechanisms of resistance in Daphnia magna exposed to the insecticide fenitrothion.
2007,
Pubmed den Besten,
Bioaccumulation and biomarkers in the sea star Asterias rubens (Echinodermata: Asteroidea): a North Sea field study.
2001,
Pubmed Denoyelle,
Cholinesterase activity as a biomarker of pesticide exposure in Allolobophora chlorotica earthworms living in apple orchards under different management strategies.
2007,
Pubmed Diamantino,
Characterization of cholinesterases from Daphnia magna straus and their inhibition by zinc.
2003,
Pubmed Duquesne,
Biochemical, metabolic, and behavioural responses and recovery of Daphnia magna after exposure to an organophosphate.
2010,
Pubmed Duquesne,
Effects of an organophosphate on Daphnia magna at suborganismal and organismal levels: implications for population dynamics.
2006,
Pubmed Garabrant,
Cholinesterase inhibition in chlorpyrifos workers: Characterization of biomarkers of exposure and response in relation to urinary TCPy.
2009,
Pubmed Guilhermino,
Inhibition of acetylcholinesterase activity as effect criterion in acute tests with juvenile Daphnia magna.
1996,
Pubmed Hackenberger,
Effect of temephos on cholinesterase activity in the earthworm Eisenia fetida (Oligochaeta, Lumbricidae).
2008,
Pubmed Hall,
The acetylcholinesterase gene of Anopheles stephensi.
1991,
Pubmed Hill,
Sex and storage affect cholinesterase activity in blood plasma of Japanese quail.
1989,
Pubmed Jemec,
The applicability of acetylcholinesterase and glutathione S-transferase in Daphnia magna toxicity test.
2007,
Pubmed Jiang,
Recombinant expression and biochemical characterization of the catalytic domain of acetylcholinesterase-1 from the African malaria mosquito, Anopheles gambiae.
2009,
Pubmed Kaufer,
Anticholinesterases induce multigenic transcriptional feedback response suppressing cholinergic neurotransmission.
1999,
Pubmed Khattab,
Immunoassays for avian butyrylcholinesterase: Implications for ecotoxicological testing and clinical biomonitoring.
2007,
Pubmed Kondo,
Enzyme-linked immunosorbent assay (ELISA) for Aleuria aurantia lectin-reactive serum cholinesterase to differentiate liver cirrhosis and chronic hepatitis.
1995,
Pubmed Li,
Hormetic response of cholinesterase from Daphnia magna in chronic exposure to triazophos and chlorpyrifos.
2011,
Pubmed Liu,
Altered quantities and in vivo activities of cholinesterase from Daphnia magna in sub-lethal exposure to organophosphorus insecticides.
2012,
Pubmed Monserrat,
Some kinetic and toxicological characteristics of thoracic ganglia cholinesterase of Chasmagnathus granulata (Decapoda, Grapsidae).
1998,
Pubmed Phillips,
Environmental, biological, and methodological factors affecting cholinesterase activity in walleye (Stizostedion vitreum).
2002,
Pubmed Printes,
Laboratory measurements of biomarkers and individual performances in Chironomus xanthus to evaluate pesticide contamination of sediments in a river of southeastern Brazil.
2011,
Pubmed Printes,
Clonal variation in acetylcholinesterase biomarkers and life history traits following OP exposure in Daphnia magna.
2008,
Pubmed Sáenz,
Biochemical biomarkers and metals in Perna perna mussels from mariculture zones of Santa Catarina, Brazil.
2010,
Pubmed Scaps,
Acetylcholinesterase activity of the polychaete Nereis diversicolor: effects of temperature and salinity.
2000,
Pubmed Stien,
Evaluation of biomarkers in caged fishes and mussels to assess the quality of waters in a bay of the NW Mediterranean Sea.
1998,
Pubmed Sturm,
Altered cholinesterase and monooxygenase levels in Daphnia magna and Chironomus riparius exposed to environmental pollutants.
1999,
Pubmed van Oosterom,
Investigation of the mud crab (Scylla serrata) as a potential bio-monitoring species for tropical coastal marine environments of Australia.
2010,
Pubmed Xuereb,
Acetylcholinesterase activity in Gammarus fossarum (Crustacea Amphipoda) Intrinsic variability, reference levels, and a reliable tool for field surveys.
2009,
Pubmed Xuereb,
Cholinesterase activity in Gammarus pulex (Crustacea Amphipoda): characterization and effects of chlorpyrifos.
2007,
Pubmed Yang,
Purification and studies on characteristics of cholinesterases from Daphnia magna.
2013,
Pubmed
