ISSN : 1225-3480
제주배꼽달팽이 (A. chejuensis) 의 metallothionein 유전자는 288 bp의 염기서열로 구성되어 있으며, 96개의 아미노산으로 이루어져 있다. 연체동물의 MT 특이적 서열 공식인 C-x-C-x (3) -C-T-G-x (3) -C-x-C-x (3) -C-x-C-K 에 대입해 보았을 때 알려진 공식과 일치하는 것을 확인하였다. BLAST 결과를 토대로 23종의 MT 서열을 얻었으며, 선행된 연구에서 사용된 5종의 MT 서열을 인위적으로 추가하여 data set를 구성하였다. ClustalX를 통해 multiple alignment하고, Neighbor-Joining 방법을 통해 phylogenetic tree를 그려본 결과 제주배꼽달팽이 (A. chejuensis) 의 MT 서열은 병안목(Stylommatophora) 에 속하는 A. quelpartensis, N. samarangae, H. pomatia, H. aspersa, S. myomphala 와 같은 그룹으로 묶이는 것을 확인 할 수 있었다. 이러한 근거를 토대로 MT 유전자 서열에 대한 연구들이 앞으로 더 많은 연체동물을 대상으로 수행된다면, 추후 분자계통학적 및 계통발생학적 방법의 연구에 유용하게 사용될 수 있는 마커로서의 충분한 가능성을 보인다는 것을 확인시켜 주었다.
Metallothioneins (MT) are cysteine-rich small proteins known for their high affinity binding to metal ions. MT confers cytoprotection against oxidative stress and acts as an efficient modulator of immunity and infection. There have been ever-increasing reports on the function of MT in environmental monitoring programmes. Lately, MT has been studied as a biomarker towards the taxonomic clustering of various species, including the molluscans. The Korean endemic land snail, Aegista chejuensis is native to Jeju Island and South coastal regions of Korea. Functional genomics information for the species is limited, although the Illumina-based DNAseq and transcriptome information are registered with the Sequence Read Archive (SRA) of National Center for Biotechnology Information (NCBI). While annotating the ORF-sensitive unigenes from the transcriptome, a MT sequence comprising of 288 bp coding region and 96 amino acid residues translatable protein was deciphered. In silico analysis using ClustalX and the MEGA7 program grouped A. chejuensis under the Stylommatophora clade of eupulmonate mollusk families. The study was able to confirm the evolutionary position of the species and established the MT sequence as a candidate for taxonomic classification of the species.
Baek, M.K., Lee, J.S., Kang, S.W., Lee, J.B., Kang, H.J., Jo, Y.H., Noh, M.Y., Han, Y.S., Choi, S.H., Chae, S.H., Park, H.S., Lee, J.S., and Lee, Y.S. (2009) Phylogenetic Analysis based on Metallothionein Gene Sequence of an Indigenous Species Pisidium (Neopisidium) coreanum in Korea. The Korean Journal of Malacology, 25: 153-160.
Barsyte, D., White, K.N., and Lovejoy, D.A. (1999) Cloning and characterization of metallothionein cDNAs in the mussel Mytilus edulis L. digestive gland. Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology, 122: 287-296.
Binz, P.-A., and Kagi, J.H.R. (1999) Metallothionein: Molecular evolution and classification. In; Klaassen CD (ed) Metallothionein IV. Birkhauser Basel. pp 7-13
Carpene, E., Andreani, G., and Isani, G. (2007) Metallothionein functions and structural characteristics. Journal of Trace Elements in Medicine and Biology 21 Suppl., 1: 35-39.
Chung, J.M., Hwang, H.J., Min, H.R., Park, J.E., Sang, M.K., Park, S.Y., Park, Y.S., Noh, M.Y., Jo, Y.H., Han, Y.S., Lee, J.S., Park, S.H., Kang, S.W., Kang, C.S., and Lee, Y.S. (2017) Molecular Phylogenetic Analysis based on Metallothionein Gene Sequence of an Endangered Species Cristaria plicata in Korea. The Korean Journal of Malacology, 33: 35-40.
Cornelia, Ileana, F., Liliana, and Lavinia, R. (2017) Metallothioneins, Saccharomyces cerevisiae, and Heavy Metals: A Biotechnology Triad? InTech: pp21-39.
Curtis D. Klaassen, Jie Liu, and Choudhuri, S. (1999) METALLOTHIONEIN, An Intracellular Protein to Protect Against Cadmium Toxicity. Toxicol., 39: 267-294.
Faria, S.C., Klein, R.D., Costa, P.G., Crivellaro, M.S., Santos, S., Bueno, S.L.S., and Bianchini, A. (2018) Phylogenetic and environmental components of inter-specific variability in the antioxidant defense system in freshwater anomurans Aegla (Crustacea, Decapoda). Scientific Reports, 8: 2850.
Ferrante, M., Vassallo, M., Mazzola, A., Brundo, M.V., Pecoraro, R., Grasso, A., and Copat, C. (2018) In vivo exposure of the marine sponge Chondrilla nucula Schmidt, 1862 to cadmium (Cd), copper (Cu) and lead (Pb) and its potential use for bioremediation purposes. Chemosphere, 193: 1049-1057.
Haas, B.J., Papanicolaou, A., Yassour, M., Grabherr, M., Blood, P.D., Bowden, J., Couger, M.B., Eccles, D., Li, B., Lieber, M., MacManes, M.D., Ott, M., Orvis, J., Pochet, N., Strozzi, F., Weeks, N., Westerman, R., William, T., Dewey, C.N., Henschel, R., LeDuc, R.D., Friedman, N., and Regev, A. (2013) De novo transcript sequence reconstruction from RNA-seq using the Trinity platform for reference generation and analysis. Nature Protocols, 8: 1494-1512.
Huang, C.W., Lee, Y.C., Lin, S.M., and Wu, W.L. (2014) Taxonomic revision of Aegista subchinensis (Mollendorff, 1884) (Stylommatophora, Bradybaenidae) and a description of a new species of Aegista from eastern Taiwan based on multilocus phylogeny and comparative morphology. Zookeys: 445: 31-55.
Jo, Y.H., Baek, M.K., Kang, S.W., Lee, J.B., Byun, I.S., Choi, S.H., Chae, S.H., Kang, J.H., Han, Y.S., Park, H.S., and Lee, Y.S. (2009) Molecular cloning and expression pattern of Metallothionein Gene from the left-handed shell, Physa acuta. The Korean Journal of Malacology, 25: 223-230.
Joshi, N., and Fass, J. (2011) Sickle: a sliding-window, adaptive, quality-based trimming tool for FastQ files. (https://github.com/najoshi/sickle)
Kang, S.W., Hwang, H.J., Park, S.Y., Wang, T.H., Park, E.B., Lee, T.H., Hwang, U.W., Lee, J.-S., Park, H.S., Han, Y.S., Lim, C.E., Kim, S., and Lee, Y.S. (2014) Mollusks Sequence Database: Version II. The Korean Journal of Malacology, 30: 429-431.
Kang, S.W., Patnaik, B.B., Hwang, H.J., Park, S.Y., Wang, T.H., Park, E.B., Chung, J.M., Song, D.K., Patnaik, H.H., Lee, J.B., Kim, C., Kim, S., Park, H.S., Lee, J.S., Han, Y.S., and Lee, Y.S. (2016) De novo Transcriptome Generation and Annotation for Two Korean Endemic Land Snails, Aegista chejuensis and Aegista quelpartensis, Using Illumina Paired-End Sequencing Technology. International Journal of Molecular Sciences, 17: 379.
Kumar, S., Stecher, G., and Tamura, K. (2016) MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Molecular Biology and Evolution, 33: 1870-1874.
L. Gnatyshyna, H. I. Falfushynska, V. V. Martyniuk, V. V. Mykhalska, Z. Yu. Kubashok, N. Y. Mischuk, A. V. Kharchuk, I. M. Sokolova, I. M. Maletska, I. V. Soltys, and Stoliar, O.B. (2017) Biochemical responses of bivalve mollusk Unio tumidus to the effect of nanoform of zinc oxide depending on the thermal regime. Біологічні студіі, 11: 25-32.
Larkin, M.A., Blackshields, G., Brown, N.P., Chenna, R., McGettigan, P.A., McWilliam, H., Valentin, F., Wallace, I.M., Wilm, A., Lopez, R., Thompson, J.D., Gibson, T.J., and Higgins, D.G. (2007) Clustal W and Clustal X version 2.0. Bioinformatics, 23: 2947-2948.
Larsson, J., Smolarz, K., Swiezak, J., Turower, M., Czerniawska, N., and Grahn, M. (2018) Multi biomarker analysis of pollution effect on resident populations of blue mussels from the Baltic Sea. Aquatic Toxicology, 198: 240-256.
Lee, J.S., and Son, M.H. (2012) Red Data Book of Endangered Mollusks in KOREA. National Institute of Biological Resources. pp152-153.
Lee, J.S, Min, B.J., Kang, S.W., Lee, J.B., Baek, M.K., Hwang, S.Y., Kim, S.H., Kho, W.G., Choi, S.H., Chae, S.H., Park, H.S., Han, Y.S., Lee, J.S., Jeong, K.H., and Lee, Y.S. (2008) Molecular Phylogenetic Study of Nesiohelix samarangae Based on Metallothionein Gene. The Korean Journal of Malacology, 24: 73-80.
Lozupone, C., and Knight, R. (2005) UniFrac: a new phylogenetic method for comparing microbial communities. Applied and Environmental Microbiology, 71: 8228-8235.
Martin, M. (2011) Cutadapt removes adapter sequences from high-throughput sequencing reads EMBnet. journal, 17: 10-12
Min, D.k. (2004) Mollusks in Korea. Min Molluscan Research Institute: pp361.
Olson, S.A. (2002) Emboss opens up sequence analysis. Briefings in bioinformatics, 3: 87-91.
Orpinell, G.C. (2017) Metallothionein family. the multipurpose protein. Influence of Mt1 in the Tg2576 mouse model of Alzeimer's disease. Universitat Autonoma de Barcelona. pp10-41
Peng, W., Li, X., Xiao, S., and Fan, W. (2018) Review of remediation technologies for sediments contaminated by heavy metals. Journal of Soils and Sediments, 18: 1701-1719.
Pertea, G., Huang, X., Liang, F., Antonescu, V., Sultana, R., Karamycheva, S., Lee, Y., White, J., Cheung, F., Parvizi, B., Tsai, J., and Quackenbush, J. (2003) TIGR Gene Indices clustering tools (TGICL): a software system for fast clustering of large EST datasets. Bioinformatics, 19: 651-652.
R.C. Braun, K.T. Pedretti*, T.L. Casavant, T.E. Scheetz, C.L. Birkett, and Roberts, C.A. (2001) Parallelization of local BLAST service on workstation clusters. Future Generation Computer Systems, 17: 745-754.
Ragusa, M.A., Nicosia, A., Costa, S., Cuttitta, A., and Gianguzza, F. (2017) Metallothionein Gene Family in the Sea Urchin Paracentrotus lividus: Gene Structure, Differential Expression and Phylogenetic Analysis. International Journal of Molecular Sciences, 18: 812.
Sang, M.K., Kang, S.W., Hwang, H.-J., Chung, J.M., Song, D.K., Min, H.R., Park, J.E., Ha, H.C., Lee, H.J., Hong, C.E., Ahn, Y.M., Park, S.Y., Park, Y.-S., Park, H.S., Han, Y.S., Lee, J.S., and Lee, Y.S. (2016) Molecular Phylogenetic Study of the Endangered Land Snail Satsuma myomphala Based on Metallothionein Gene. The Korean Journal of Malacology, 32: 263-268.
Worthing, K.A., Abraham, S., Coombs, G.W., Pang, S., Saputra, S., Jordan, D., Trott, D.J., and Norris, J.M. (2018) Clonal diversity and geographic distribution of methicillin-resistant Staphylococcus pseudintermedius from Australian animals: Discovery of novel sequence types. Veterinary Microbiology, 213: 58-65.
