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Cloning and characterization of the calreticulin gene in Asian seabass (Lates calcarifer)

Published online by Cambridge University Press:  17 November 2011

Z. Y. Bai ,
Z. Y. Zhu ,
C. M. Wang ,
J. H. Xia ,
X. P. He  and
G. H. Yue
Z. Y. Bai
Affiliation:
Temasek Life Sciences Laboratory, Molecular Population Genetics Group, National University of Singapore, 1 Research Link 117604, Singapore Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, College of Fisheries and Life Science, Shanghai Ocean University, Ministry of Education, Shanghai 201306, China
Z. Y. Zhu
Affiliation:
Temasek Life Sciences Laboratory, Molecular Population Genetics Group, National University of Singapore, 1 Research Link 117604, Singapore
C. M. Wang
Affiliation:
Temasek Life Sciences Laboratory, Molecular Population Genetics Group, National University of Singapore, 1 Research Link 117604, Singapore
J. H. Xia
Affiliation:
Temasek Life Sciences Laboratory, Molecular Population Genetics Group, National University of Singapore, 1 Research Link 117604, Singapore
X. P. He
Affiliation:
Temasek Life Sciences Laboratory, Molecular Population Genetics Group, National University of Singapore, 1 Research Link 117604, Singapore
G. H. Yue*
Affiliation:
Temasek Life Sciences Laboratory, Molecular Population Genetics Group, National University of Singapore, 1 Research Link 117604, Singapore
*
†E-mail: genhua@tll.org.sg
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Abstract

Calreticulin (CRT) is a Ca2+-binding molecular chaperone in the endoplasmic reticulum. We cloned and characterized the CRT gene in an important marine food fish species Asian seabass (Lates calcarifer). The full-length DNA of the CRT gene was 2194 bp, including a complete open reading frame encoding 420 amino acid residues, a 113 bp 5′-untranslated region and an 818 bp 3′-untranslated region. The CRT gene contained nine exons and eight introns covering a total of 2772 bp genomic DNA from the start to stop codon. Ten single nucleotide polymorphisms (SNPs) were detected in introns and an exon in six individuals collected from five different locations. The CRT gene was assigned to linkage group 4 of the linkage map of Asian seabass. Quantitative real-time PCR revealed that the CRT gene was highly expressed in liver at the age of 1, 3 and 7 months under normal conditions, whereas its expression in liver reduced sharply after 0.5 to 2 h cold challenge at 16°C, and then increased slowly. A preliminary association analysis showed a significant (P < 0.001) association between the SNP6 in the CRT gene and the mortality after cold challenge at 16°C. Our results suggest that the CRT gene is associated with cold tolerance of Asian seabass and further investigation will be necessary to illustrate the underlying mechanisms.

Keywords

Barramundipolymorphismcold toleranceQTL
Type
Full Paper
Information
animal , Volume 6 , Issue 6 , June 2012 , pp. 887 - 893
Copyright
Copyright © The Animal Consortium 2011

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References

Chen, F, Hayes, PM, Mulrooney, DM, Pan, A 1994. Identification and characterization of cDNA clones encoding plant calreticulin in barley. Plant Cell 6, 835–843.Google Scholar
Chou, R, Lee, HB 1997. Commercial marine fish farming in Singapore. Aquaculture Research 28, 767–776.Google Scholar
Danzmann, RG, Jackson, TR, Ferguson, MM 1999. Epistasis in allelic expression at upper temperature tolerance QTL in rainbow trout. Aquaculture 173, 45–58.Google Scholar
De Santis, C, Evans, BS, Smith-Keune, C, Jerry, DR 2008. Molecular characterization, tissue expression and sequence variability of the barramundi (Lates calcarifer) myostatin gene. BMC Genomics 9, 82.CrossRefGoogle ScholarPubMed
Ferreira, CA, Da Silva Vaz, I, da Silva, SS, Haag, KL, Valenzuela, JG, Masuda, A 2002. Cloning and partial characterization of a Boophilus microplus (Acari: Ixodidae) calreticulin. Experimental Parasitology 101, 25–34.CrossRefGoogle ScholarPubMed
Fliegel, L, Burns, K, MacLennan, DH, Reithmeier, RA, Michalak, M 1989. Molecular cloning of the high affinity calcium-binding protein (calreticulin) of skeletal muscle sarcoplasmic reticulum. The Journal of Biological Chemistry 264, 21522–21528.CrossRefGoogle ScholarPubMed
Food and Agriculture Organization (FAO) 2008. Food and Agriculture Organization of the United Nations statistics 2008. www.fao.org/fishery/culturedspecies/Lates_calcarifer/enGoogle Scholar
Frost, LA, Evans, BS, Jerry, DR 2006. Loss of genetic diversity due to hatchery culture practices in barramundi (Lates calcarifer). Aquaculture 261, 1056–1064.Google Scholar
Guiguen, Y, Cauty, C, Fostier, A, Fuchs, J, Jalabert, B 1994. Reproductive cycle and sex inversion of the seabass, Lates calcarifer, reared in sea cages in French Polynesia: histological and morphometric description. Environmental Biology of Fishes 39, 231–247.Google Scholar
Hojrup, P, Roepstorff, P, Houen, G 2001. Human placental calreticulin characterization of domain structure and post-translational modifications. European Journal of Biochemistry 268, 2558–2565.CrossRefGoogle ScholarPubMed
Jia, XY, Xu, CY, Jing, RL, Li, RZ, Mao, XG, Wang, JP, Chang, XP 2008. Molecular cloning and characterization of wheat calreticulin (CRT) gene involved in drought-stressed responses. Journal of Experimental Botany 59, 739–751.CrossRefGoogle ScholarPubMed
Kales, S, Fujiki, K, Dixon, B 2004. Molecular cloning and characterization of calreticulin from rainbow trout (Oncorhynchus mykiss). Immunogenetics 55, 717–723.CrossRefGoogle ScholarPubMed
Komatsu, S, Yang, GX, Khan, M, Onodera, H, Toki, S, Yamaguchi, M 2007. Over-expression of calcium-dependent protein kinase 13 and calreticulin interacting protein 1 confers cold tolerance on rice plants. Molecular Genetics & Genomics 277, 713–723.Google Scholar
Lander, ES, Green, P, Abrahamson, J, Barlow, A, Daly, MJ, Lincoln, SE, Newburg, L 1987. MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1, 174–181.Google Scholar
Li, ZG, Cao, YR, Zhang, JS, Chen, SY 2008. Characterization of Arabidopsis calreticulin mutants in response to calcium and salinity stresses. Progress in Natural Science 18, 1219–1224.Google Scholar
Michalak, M, Corbett, EF, Mesaeli, N, Nakamura, K, Opas, M 1999. Calreticulin: one protein, one gene, many functions. Biochemical Journal 344, 281–292.Google Scholar
Nelson, JS 2006. Fishes of the world. John Wiley and Sons, Inc., New York.Google Scholar
O'Sullivan, D 2004. Australian barra for New York. Australia Aquaculture 18, 40–43.Google Scholar
Ostwald, TJ, Mclennan, DH 1974. Isolation of a high affinity calcium-binding protein from sarcoplasmic reticulum. The Journal of Biology Chemistry 249, 974–979.Google Scholar
Perrone, L, Tell, G, Di Lauro, R 1999. Calreticulin enhances the transcriptional activity of thyroid transcription factor-1 by binding to its homeodomain. The Journal of Biological Chemistry 274, 4640–4645.Google Scholar
Porcellini, S, Traggiai, E, Schenk, U, Ferrera, D, Matteoli, M, Lanzavecchia, A, Michalak, M, Grassi, F 2006. Regulation of peripheral T cell activation by calreticulin. The Journal of Experimental Medicine 203, 461–471.CrossRefGoogle ScholarPubMed
Rokeach, LA, Haselby, JA, Meilof, JF, Smeenk, RJ, Unnasch, TR, Greene, BM, Hoch, SO 1991. Characterization of the autoantigen calreticulin. The Journal of Immunology 147, 3031–3039.Google Scholar
Tucker, JW, Russell, DJ, Rimmer, M 2002. Barramundi culture: a success story for aquaculture in Asia and Australia. World Aquaculture 33, 67–72.Google Scholar
Van Ooijen, JW, Boer, MP, Jansen, RC 2002. MapQTL 4.0: software for the calculation of QTL positions on genetic maps. Plant Research International, Wageningen, the Netherlands.Google Scholar
Volvich, L, Appelbaum, S 2001. Length to weight relationship of sea bass Lates calcarifer (Bloch) reared in a closed recirculating system. Israeli Journal of Aquaculture – Bamidgeh 53, 158–163.Google Scholar
Wang, CM, Lo, LC, Zhu, ZY, Yue, GH 2006. Genome-scan QTL for growth-related traits in an F1 family from a breeding population of Asian seabass. BMC Genomics 7, 274.Google Scholar
Wang, CM, Zhu, ZY, Lo, LC, Feng, F, Lin, G, Yang, WT, Li, J, Yue, GH 2007. A microsatellite linkage map of Barramundi, Lates calcarifer. Genetics 175, 907–915.Google Scholar
Wang, CM, Lo, LC, Zhu, ZY, Lin, G, Feng, F, Li, J, Yang, WT, Tan, J, Chou, R, Lim, HS, Orban, L, Yue, GH 2008. Estimating reproductive success of brooders and heritability of growth traits in Asian seabass using microsatellites. Aquaculture Research 39, 1612–1619.Google Scholar
Waser, M, Mesaeli, N, Spencer, C, Michalak, M 1997. Regulation of calreticulin gene expression by calcium. Journal Cell Biology 138, 547–557.CrossRefGoogle ScholarPubMed
Xu, YX, Zhu, ZY, Lo, LC, Wang, CM, Lin, G, Feng, F, Yue, GH 2006. Characterization of two parvalbumin genes and their association with growth traits in Asian seabass (Lates calcarifer). Animal Genetics 37, 266–268.Google Scholar
Yowe, DL, Epping, RJ 1995. Cloning of the barramundi growth hormone-encoding gene – a comparative-analysis of higher and lower vertebrate GH genes. Gene 162, 255–259.Google Scholar
Yue, GH, Orban, L 2005. A simple and affordable method for high throughput DNA extraction from animal tissues for PCR. Electrophoresis 26, 3081–3083.Google Scholar
Yue, G, Li, Y, Orban, L 2001. Characterization of microsatellites in the IGF-2 and GH genes of Asian seabass (Lates calcarifer). Marine Biotechnology 3, 1–3.Google Scholar
Yue, GH, Li, Y, Chao, TM, Chou, R, Orban, L 2002. Novel microsatellites from Asian sea bass (Lates calcarifer) and their application to broodstock analysis. Marine Biotechnology 4, 503–511.CrossRefGoogle ScholarPubMed
Yue, GH, Zhu, ZY, Lo, LC, Wang, CM, Lin, G, Feng, F, Pang, HY, Li, J, Liu, HM, Gong, P, Tan, J, Lim, HS, Chou, R, Orban, L 2009. Genetic variation and population structure of Asian seabass (Lates calcarifer) in the Asia-Pacific region. Aquaculture 293, 22–28.Google Scholar
Zhu, ZY, Lin, G, Lo, LC, Xu, YX, Renee, C, Yue, GH 2006. Genetic analyses of Asian seabass stocks using novel polymorphic microsatellites. Aquaculture 256, 167–173.Google Scholar
Supplementary material: File

Bai Supplementary Figure

Fig. S1 Nucleotide and deduced amino acid sequence of seabass calreticulin(CRT) cDNA

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