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Molecular cloning and expression pattern of Metallothionein Gene from the left-handed shell, Physa acuta

Abstract

Metallothioneins (MTs) play a key role in metallic homeostasis and detoxification in most living organisms. In an attempt to study the biological functions and significance of MT in a snail, we cloned and partially characterized the MT gene from the left-handed snail, Physa acutawhich has been regarded as a potential biomonitering species for fresh water. The complete cDNA sequence of PaMT cDNA was identified from the expressed sequence tag (EST) sequencing project of Physa acuta. The coding region of 180 bp gives 60 amino acid residues including the initiation methionine and termination codon. Clustering and phylogenic analysis of PaMT with other MT amino acid sequences show that it has some identities to Helix pomatia (60%), Arianta arbustorum (58%), Perna viridis (49%), Mytilus edulis (49%), Bathymodiolus azoricus (49%), Bathymodiolus azoricus (48%) and Bathymodiolus sp. FD-2002 (48%). Time dependent induction for PaMT from P. acuta exposed with cadmium (50 ppb) indicated that PaMT was induced at 4-8 hr after exposure. It remains to further develop PaMTas a potential biomarker for water contamination in fresh water.

keywords
cloning, metallothionein, Physa acuta, bio-monitoring

Reference

1.

(1990). . Veterinary & Human Toxicology, 32, 212-216.

2.

Takimoto, Y;Ohshima, M;Miyamoto, J. (1987). Comparative metabolism of fenitrothion in aquatic organisms. II. Metabolism in the freshwater snails, Cipangopaludina japonica and Physa acuta.. Ecotoxicology and Environmental Safety, 13(1), 118-125. 10.1016/0147-6513(87)90049-2.

3.

Tamura, K.;Dudley, J.;Nei, M.;Kumar, S.. (2007). MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) Software Version 4.0. Molecular Biology and Evolution, 24(8), 1596-1599. 10.1093/molbev/msm092.

4.

(1961). . The Journal of biological chemistry, 236, 2435-2442.

5.

Kojima, Y;Berger, C;Vallee, B L;Kägi, J H. (1976). Amino-acid sequence of equine renal metallothionein-1B.. Proceedings of the National Academy of Sciences, 73(10), 3413-3417. 10.1073/pnas.73.10.3413.

6.

(2008). . Korean Journal of Malacology, 24, 73-80.

7.

(2006). . Korean Journal of Malacology, 22, 83-91.

8.

(2004). . Korean Journal of Malacology, 20, 165-169.

9.

(2002). . Korean Journal of Malacology, 18, 15-21.

10.

(2006). . Korean Journal of Malacology, 22, 157-165.

11.

Leung, K.M.Y.;Morley, N.J.;Grist, E.P.M.;Morritt, D.;Crane, M.. (2004). Chronic toxicity of tributyltin on development and reproduction of the hermaphroditic snail Physa fontinalis: Influence of population density. Marine Environmental Research, 58(2), 157-162. 10.1016/j.marenvres.2004.03.011.

12.

Maltez, H.F.;Tagle, M.V.;Fernandez de la Campa, M.d.R.;Sanz-Medel, A.. (2009). Metal-metallothioneins like proteins investigation by heteroatom-tagged proteomics in two different snails as possible sentinel organisms of metal contamination in freshwater ecosystems. Analytica Chimica Acta, 650(2), 234-240. 10.1016/j.aca.2009.07.052.

13.

(1994). . Journal of molecular evolution, 38, 300-304.

14.

(1998). . Genome Research, 8, 186-194.

15.

Johnson, M.;Zaretskaya, I.;Raytselis, Y.;Merezhuk, Y.;McGinnis, S.;Madden, T. L.. (2008). NCBI BLAST: a better web interface. Nucleic Acids Research, 36(suppl2), W5-W9. 10.1093/nar/gkn201.

16.

(1998). . Genome Research, 8, 175-185.

17.

Felsenstein, Joseph. (1985). Confidence Limits on Phylogenies: An Approach Using the Bootstrap. Evolution, 39(4), 783-791. 10.2307/2408678.

18.

Huang, X;Madan, A. (1999). CAP3: A DNA sequence assembly program.. Genome Research, 9(9), 868-877. 10.1101/gr.9.9.868.

19.

(1960). . The Journal of biological chemistry, 235, 3460-3465.

20.

Canivet, V.;Chambon, P.;Gibert, J.. (2001). Toxicity and Bioaccumulation of Arsenic and Chromium in Epigean and Hypogean Freshwater Macroinvertebrates. Archives of Environmental Contamination and Toxicology, 40(3), 345-354. 10.1007/s002440010182.

21.

(2001). . Ocean and Polar Research, 23, 337-345.

22.

Bebianno, M. J.;Serafim, M. A.. (2003). Variation of Metal and Metallothionein Concentrations in a Natural Population of Ruditapes decussatus. Archives of Environmental Contamination and Toxicology, 44(1), 53-66. 10.1007/s00244-002-2004-7.

23.

(1990). . Journal of molecular biology, 215, 403-410.

24.

Azevedo, J. S.;Serafim, A.;Company, R.;Braga, E. S.;F&aacute;varo, D. I.;Bebianno, M. J.. (2009). Biomarkers of exposure to metal contamination and lipid peroxidation in the benthic fish <I>Cathorops spixii</I> from two estuaries in South America, Brazil. Ecotoxicology, 18(8), 1001-1010. 10.1007/s10646-009-0370-x.

25.

BACCHETTA. (2002). . Journal Molluscan Studies, 68(2), 181-186. 10.1093/mollus/68.2.181.

26.

Tanguy, A.;Moraga, D.. (2001). Cloning and characterization of a gene coding for a novel metallothionein in the Pacific oyster Crassostrea gigas (CgMT2): a case of adaptive response to metal-induced stress?. Gene, 273(1), 123-130. 10.1016/S0378-1119(01)00577-7.

27.

Cuppen, Jan G. M.;Crum, Steven J. H.;Van den Heuvel, Harry H.;Smidt, Rob A.;Van den Brink, P. (2002). Effects of a Mixture of Two Insecticides in Freshwater Microcosms&colon; I. Fate of Chlorpyrifos and Lindane and Responses of Macroinvertebrates. Ecotoxicology, 11(3), 165-180. 10.1023/A:1015470731330.

28.

Braun, R.C.;Pedretti, K.T.;Casavant, T.L.;Scheetz, T.E.;Birkett, C.L.;Roberts, C.A.. (2001). Parallelization of local BLAST service on workstation clusters. Future Generation Computer Systems, 17(6), 745-754. 10.1016/S0167-739X(00)00057-1.

29.

Butler, R.A.;Roesijadi, G.. (2001). Quantitative reverse transcription polymerase chain reaction of a molluscan metallothionein mRNA. Aquatic Toxicology, 54(1), 59-67. 10.1016/S0166-445X(00)00176-4.

30.

(2009). . Korean Journal of Malacology, 25, 153-160.

31.

Tanguy, A.;Mura, C.;Moraga, D.. (2001). Cloning of a metallothionein gene and characterization of two other cDNA sequences in the Pacific oyster Crassostrea gigas (CgMT1). Aquatic Toxicology, 55(1), 35-47. 10.1016/S0166-445X(01)00160-6.

32.

Zdobnov, Evgeni M.;Apweiler, Rolf. (2001). InterProScan &ndash; an integration platform for the signature-recognition methods in InterPro. Bioinformatics, 17(9), 847-848. 10.1093/bioinformatics/17.9.847.

33.

34.

(1987). . Molecular biology and evolution, 4, 406-425.

35.

Mourgaud, Yohan;Martinez, &Eacute;ric;Geffard, Alain;Andral, Bruno;Stanisiere, Jean-Yves;A. (2002). Metallothionein concentration in the mussel Mytilus galloprovincialis as a biomarker of response to metal contamination: validation in the field. Biomarkers, 7(6), 479-490. 10.1080/1354750021000034528.

36.

Nebeker, A V;Schuytema, G S. (1998). Chronic effects of the herbicide diuron on freshwater cladocerans, amphipods, midges, minnows, worms, and snails.. Archives of Environmental Contamination and Toxicology, 35(3), 441-446. 10.1007/s002449900400.

37.

38.

Simes, D.C.;Bebianno, M.J.;Moura, J.J.G.. (2003). Isolation and characterisation of metallothionein from the clam Ruditapes decussatus. Aquatic Toxicology, 63(3), 307-318. 10.1016/S0166-445X(02)00185-6.

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