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| Cloning of MIPS Gene in Large Yellow Croaker (Larimichthys crocea) and the Expression Analysis Under Cold Treatments |
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Abstract Myo-inositol-1-phosphate synthase (MIPS) is one of the key enzyme in biological inositol metabolism. In order to explore the role of MIPS in the stress response of Larimichthys crocea under cold treatments, MIPS gene was cloned and then its sequence were analyzed and the expression changes were detected under chronic and acute cold stress. The full length of MIPS cDNA (GenBank accession No. MG760436) was 2 599 bp which contained a 1 659 bp open reading frame (ORF) and encoded 553 amino acids, the molecular weight of putative MIPS protein was 60.5 kD. Sequence alignment showed that the amino acid sequence of L. crocea MIPS shared high similarity (85.5%~88.4%) to other teleost and all of these MIPS had 4 highly conserved core regions. In the phylogenetic tree, L. crocea MIPS gathered with MIPS of other teleost and was most closely related to the MIPS of Fugu rubripes (Takifugu rubripes). During chronic cold stress (water temperature slowed decreased from 12 ℃ to 6 ℃), the MIPS expression in gill, skin and heart significantly increased at first and then significantly decreased (P＜0.05), and the maximum appeared at 8 ℃ which were respectively 13.71, 89.50 and 50.83 fold, respectively, compared to 12 ℃. The expression continuously elevated in brain and muscle during chronic cold stress and at 6 ℃ the expression were 99.89 and 110.17 fold higher than before cold stress. And there were no significant changes in intestine, kidney, liver and spleen (P＞0.05). During acute cold stress (from 12 ℃ to 8 ℃ immediately and keeping 8 ℃ for 4 h), the expression level of MIPS were significantly up-regulated (P＜0.05) in all the detected tissues except intestine. The 2 biggest expression changes were appeared in brain and muscle, which were 54.53 and 32.89 fold higher than before cold stress. The expression changes in both chronic and acute cold treatments indicated MIPS participates in the response to cold stress and has possible relationship to cold adaptability of L. crocea, brain and muscle are the 2 most important tissues in the response to cold stress. These results provide reference for researching the function of MIPS gene and the mechanism of response to cold stress, as well as breeding low-temperature tolerant cultivars of L. crocea.
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Received: 03 July 2017
Published: 04 February 2018
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