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A trehalose-6-phosphate synthase gene from Saccharina japonica (Laminariales, Phaeophyceae)

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Abstract

The full-length cDNA sequence of a trehalose-6-phosphate synthase gene from Saccharina japonica (designated as SjaTPS) (Accession: KC578568) was isolated based on homologous cloning and RACE-PCR. It was 4,127 bp, with 320 bp 5′-UTR, 21 bp 3′-UTR, and open reading frame (ORF) of 3,786 bp. The deduced 1,261 amino acids characterized with predicted molecular weight of 137.84 kDa and theoretical isoelectric point of 7.12. The SjaTPS had one N-terminal CBM20 (family 20 carbohydrate-binding module) domain, one TPS domain (trehalose-6-phosphate synthase) in the middle region and a single TPP (trehalose-6-phosphate phosphatase) domain near the C-terminus. Structural analysis suggested that the SjaTPS putatively functioned as trehalose-6-phosphate synthase, and might be related to laminaran metabolism in S. japonica. Homology analysis indicated that the SjaTPS shared 49–70 % similarities with the 13 known TPS sequences of other algae; only 55 % amino acid similarities were detected between SjaTPS and the previously reported TPS sequence of S. japonica (Accession: DQ666325). Phylogenetic analysis revealed close affinity between SjaTPS and TPS of brown alga Ectocarpus siliculosus (Accession: CBJ29609). Transcriptional analysis showed that desiccation greatly enhanced SjaTPS expression and the maximum appeared at 3 h, which was about 300-fold compared to that of the start, implied that SjaTPS was involved with drought adaption in kelp. In vitro expression of SjaTPS showed that one distinct band existed at ~115 kDa, and western blot detection proved that it was positive to the anti-His antibody with high specificity. Our results increased the knowledge of trehalose-6-phosphate synthase properties in S. japonica and also important for better understanding the role trehalose plays in kelp abiotic tolerance for adaption to the sublittoral habitats.

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Acknowledgments

This research was supported by National Natural Science Foundation of China (No. 40976085) and Shandong Agriculture Breeding Engineering Biological Resources Innovation of Research Project and National High Tech 863 Project (2012AA10A406). Sincerely thanks are due to Lin Xiao, Jin Zhao, Ge Liu for their help with the experiments. The authors acknowledged the anonymous reviewers for the critical comments and suggestions for the manuscript.

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  1. Chinese Academy of Sciences, Institute of Oceanology, Qingdao, 266071, China

    Yunyan Deng, Xiuliang Wang, Hui Guo & Delin Duan

  2. University of the Chinese Academy of Sciences, Beijing, 100049, China

    Hui Guo

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  1. Yunyan Deng
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Correspondence to Delin Duan.

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Deng, Y., Wang, X., Guo, H. et al. A trehalose-6-phosphate synthase gene from Saccharina japonica (Laminariales, Phaeophyceae). Mol Biol Rep 41, 529–536 (2014). https://doi.org/10.1007/s11033-013-2888-5

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