Cloning and truncation modification of trehalose-6-phosphate synthase gene from Selaginella pulvinata.

Sheng-Mei Zhao, Feng-Ling Fu, Lin Gou, Han-Guang Wang, Gang He, Wan-Chen Li

Maize Research Institute, Sichuan Agricultural University, Chengdu, Sichuan 611130, People's Republic of China.

Gene 2013 Jan 10

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A homologous sequence was amplified from resurrection plant Selaginella pulvinta by RACE technique, proved to be the full-length cDNA of trehalose-6-phosphate synthase gene by homologous alignment and yeast complementation assay, and nominated as SpTPS1 gene. The open reading frame of this gene was truncated 225bp at the 5'-end, resulting the N-terminal truncation modification of 75 amino acids for its encoding protein. The TPS1 deletion mutant strain YSH290 of the brewer's yeast transformed by the truncated gene SpTPS1Δ and its original full-length version restored growth on the medium with glucose as a sole carbon source and displayed growth curves with no significant difference, indicating their encoding proteins functioning as TPS enzyme. The TPS activity of the mutant strain transformed by the truncated gene SpTPS1Δ was about six fold higher than that transformed by its original version, reasoning that the extra N-terminal extension of the full-length amino acid sequence acts as an inhibitory domain to trehalose synthesis. However, the trehalose accumulation of the mutant strain transformed by the truncated gene SpTPS1Δ was only 8% higher than that transformed by its original version. This result is explained by the feedback balance of trehalose content coordinated by the comparative activities between trehalose synthase and trehalase. The truncated gene SpTPS1Δ is suggested to be used in transgenic operation, together with the inhibition of trehalase activity by the application of validamycin A or genetic deficiency of the endogenous trehalase gene, for the enhancement of trehalose accumulation and improvement of abiotic tolerance in transgenic plants. Copyright © 2012. Published by Elsevier B.V.

Citation

Sheng-Mei Zhao, Feng-Ling Fu, Lin Gou, Han-Guang Wang, Gang He, Wan-Chen Li. Cloning and truncation modification of trehalose-6-phosphate synthase gene from Selaginella pulvinata. Gene. 2013 Jan 10;512(2):414-21