Cloning and stage-specific expression of CK-M1 gene during metamorphosis of Japanese flounder, Paralichthys olivaceus |
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| Authors: | Yanjie Chen Quanqi Zhang Jie Qi Zhigang Wang Xubo Wang Yeying Sun Qiwang Zhong Shuo Li Chunmei Li |
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| Institution: | (1) Field Science Education and Research Center and Graduate School of Agriculture, Kyoto University, Kitashirakawa, Sakyo, Kyoto 606-8502, Japan;(2) Miyako Station, National Center for Stock Enhancement, Fisheries Research Agency, Miyako Iwate, 027-0097, Japan;(3) Present address: Seikai National Fisheries Research Institute, Coastal Fisheries and Aquaculture Division, Fisheries Research Agency, Taira, Nagasaki Nagasaki, 851-2231, Japan;(4) Present address: Stock Enhancement Unit, Fukushima Prefectural Fisheries Experimental Station, Onahama, Iwaki Fukushima, 970-0316, Japan;(5) Present address: Borneo Marine Research Institute, Universiti Malaysia Sabah, Locked Bag 2073, 88999 Kota Kinabalu, Sabah, Malaysia; |
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| Abstract: | The symmetrical body of flatfish larvae changes dramatically into an asymmetrical form after metamorphosis. The molecular mechanisms responsible for this change are poorly understood. As an initial step to clarify these mechanisms, we used representational difference analysis of cDNA for the identification of genes active during metamorphosis in the Japanese flounder, Paralichthys olicaceus. One of the up-regulated genes was identified as creatine kinase muscle type 1 (CK-M1). Sequence analysis of CK-M1 revealed that it spanned 1 708 bp and encoded a protein of 382 amino acids. The overall amino acid sequence of the CK-M1 was highly conserved with those of other organisms. CK-M1 was expressed in adult fish tissues, including skeletal muscle, intestine and gill. Whole mount in-situ hybridization showed that the enhanced expression of CK-M1 expanded from the head to the whole body of larvae as metamorphosis progressed. Quantitative analysis revealed stage-specific high expression of CK-M1 during metamorphosis. The expression level of CK-M1 increased initially and peaked at metamorphosis, decreased afterward, and finally returned to the pre-metamorphosis level. This stage-specific expression pattern suggested strongly that CK-M1 was related to metamorphosis in the Japanese flounder. Its specific role in metamorphosis requires further study. |
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