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Tan S.,Foshan University | Tan S.,South China Normal University | Li H.,Foshan University | Li H.,Holdone Aquaculture Breeding Ltd Company | And 2 more authors.
Genes and Genomics | Year: 2016

The metallothionein 2 (mt2) genomic gene with 667 bp was cloned from the grass carp (Ctenopharyngodon idellus). The structure of mt2 gene was similar with the reported vertebrae mt gene, containing three exons interrupted by two introns. Only one MRE mortify was found in the 5′ flanking region at proximal cluster which was quite different from the previous reports. The number of deduced amino acid was 60, characterized by cysteine-rich. The basal levels and the transcriptional responses of mt2 gene in different organs to cadmium (Cd) were measured using the quantitative real-time PCR (qRT-PCR). In the kidney, the constructive level of the mt2 mRNA was the highest, while the lowest expression level was detected in the skin. The induced impacts of Cd on mt2 mRNA expression were various from tissues. In the liver, gill and skin, the gene was quickly induced at 6 h. After a transient suppression, the expression levels of mt2 mRNA were increased in the intestine and spleen at 12 h, and mt2 mRNA expressed upwards in the bone at 24 h. In the kidney and bone, the expressions elevated only after 48 h Cd induction. The inducibility of mt2 gene to Cd demonstrated that it played a significant role during the metal toxicant detoxification in grass carp and the mt2 gene in the liver was most sensitive to Cd induction. © 2015, The Genetics Society of Korea and Springer-Science and Media. Source


Tan S.,Foshan University | Tan S.,Holdone Aquaculture Breeding Ltd Company | Li H.,Foshan University | Li H.,Holdone Aquaculture Breeding Ltd Company | And 3 more authors.
Ecotoxicology and Environmental Safety | Year: 2014

To gain more knowledge about the physiological regulation of metal pollutant detoxification in grass carp, we examined Cd concentration and its the potential influence on the expression of metallothionein 2 (MT2) and multidrug resistance protein 2 (ABCC2) mRNA in the liver and kidney, using in vitro and in vivo experiments. First, the full-length of MT2 cDNA and partial ABCC2 cDNA was obtained, consisting 183bp and 366bp respectively. In vivo approach, grass carp received 96h exposure of Cd (1/10 LD50), and MT2 and ABCC2 mRNA expression were determined by qRT-PCR. The Cd treatment resulted in an increase of MT2 mRNA level in the liver with Cd accumulation. Nonetheless, the elevation ABCC2 mRNA in the liver was appeared at 48h after Cd exposure, as well as the expression of MT2 and ABCC2 mRNA in the kidney. The in vitro experiment was carried out using the hepatocyte (L86) and nephroblasts (CIK). The qRT-RCR results showed that MT2 and ABCC2 mRNA dramatically increased following Cd exposure (1/10 LD50); however, ABCC2 mRNA expression was suppressed in the L86 cell line at first (6h). In conclusion, this result suggested that both MT2 and ABCC2 mRNA may play important roles in the detoxification of toxic metals, and MT2 gene was more sensitive to Cd induction. © 2014 Elsevier Inc. Source


Yu H.,Foshan University | Tan S.,Foshan University | Tan S.,Guangzhou University | Zhao H.,Foshan University | And 2 more authors.
Gene | Year: 2013

In this study, the association between MH-. DAB gene polymorphism and disease resistance was evaluated by challenging grass carp (Ctenopharyngodon idellus) with Flavobacterium columnare. Eight genotypes and six alleles were found, and named by common nomenclature. The genotypes AA, BB, EE, and DE, and the alleles Ctid-. DAB1. 0101, Ctid-. DAB1. 0201 and Ctid-. DAB1. 0401 were more preponderant in fish. The genotype BB was associated with higher resistance to F. columnare, as well as two alleles Ctid-. DAB. 0101 and Ctid-. DAB. 0201. Allele Ctid-. DAB. 0102 has decreased resistance to F. columnare. The expression of MH-. DAB gene was decreased in the liver, kidney, and intestine but not in the spleen, gill, and skin at 2. days post infection (dpi), versus to that in the control group. MH-. DAB gene expression was up-regulated in most tissues but remained at normal levels in the intestine at 15. days post infection. Our data suggested that MH-. DAB polymorphism can be used as a potential genetic marker for disease resistance breeding of grass carp in the future. © 2013 Elsevier B.V. Source

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