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Molecular cloning and characterization of alpha-class glutathioneS-transferase gene from the liver of silver carp, bighead carp, and other major chinese freshwater fishes
- Source :
- Journal of Biochemical and Molecular Toxicology. 20:114-126
- Publication Year :
- 2006
- Publisher :
- Wiley, 2006.
-
Abstract
- Two full-length cDNAs encoding glutathione S-transferase (GST) were cloned and sequenced from the hepatopancreas of planktivorous silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis). The silver carp and bighead carp GST cDNA were 920 and 978 bp in length, respectively, and both contained an open reading frame that encoding 223 amino acids. Partial GST cDNA sequences were also obtained from the liver of grass carp (Ctenopharyngodon idellus), crucian carp (Carassius auratu), mud carp (Cirrhinus molitorella), and tilapia (Oreochromis nilotica). All these GSTs could be classified as alpha-class GSTs on the basis of their amino acid sequence identity with other species. The three-dimensional structure of the silver carp GST was predicted using a computer program, and was found to fit the classical two-domain GST structure. Using the genome walker method, a 875-bp 5'-flanking region of the silver carp GST gene was obtained, and several lipopolysaccharide (LPS) response elements were identified in the promoter region of the phytoplanktivorous fish GST gene, indicating that the GST gene expression of this fish might be regulated by LPS, released from the toxic blue-green algae producing microcystins. To compare the constitutive expression level of the liver GST gene among the six freshwater fishes with completely different tolerance to microcystins, beta-actin was used as control and the ratio GST/beta-actin mRNA (%) was determined as 130.7 +/- 6.6 (grass carp), 103.1 +/- 8.9 (bighead carp), 92.6 +/- 15.0 (crucian carp), 72.3 +/- 7.8 (mud carp), 58.8 +/- 11.5 (silver carp), and 33.6 +/- 13.7 (tilapia). The constitutive expression level of the liver GST gene clearly shows that all the six freshwater fishes had a negative relationship with their tolerance to microcystins: high-resistant fishes (phytoplanktivorous silver carp and tilapia) had the lowest tolerance to microcystins and the high-sensitive fish (herbivorous grass carp) had the highest tolerance to microcystins. Taken together with the reciprocal relationship of constitutive and inducible liver GST expression level in some of the tested fish species to microcystin exposure, a molecular mechanism for different microcystin detoxification abilities of the warm freshwater fishes was discussed.
- Subjects :
- Models, Molecular
China
Carps
DNA, Complementary
5' Flanking Region
Health, Toxicology and Mutagenesis
Molecular Sequence Data
Fresh Water
Regulatory Sequences, Nucleic Acid
Toxicology
Biochemistry
Protein Structure, Secondary
Animals
Humans
Amino Acid Sequence
RNA, Messenger
Cloning, Molecular
Carp
Molecular Biology
Phylogeny
Glutathione Transferase
Silver carp
Base Sequence
Sequence Homology, Amino Acid
biology
Ecology
Sequence Analysis, DNA
General Medicine
biology.organism_classification
Bighead carp
Molecular biology
Grass carp
Cirrhinus molitorella
Glutathione S-transferase
Liver
Crucian carp
Carassius
biology.protein
Molecular Medicine
sense organs
Sequence Alignment
Subjects
Details
- ISSN :
- 10990461 and 10956670
- Volume :
- 20
- Database :
- OpenAIRE
- Journal :
- Journal of Biochemical and Molecular Toxicology
- Accession number :
- edsair.doi.dedup.....d9d3908e4dd709da06c99cb094f312ef