Cloning and functional analysis of trehalose -6-phosphate synthase gene from Selaginella pulvinata
Received date: 2009-10-09
Revised date: 2009-12-02
Online published: 2010-05-15
Trehalose-6-phosphate synthase, a key enzyme in trehalose synthesis pathway of plant, plays an important role in response to abiotec stress in xerophilous rock lily and other resurrection plants. In this study, homologous amplification and RACE technique were used to clone gene SpTPS1 for trehalose-6-phosphate synthase from Selaginella pulvinata, which is an endemic xerophilous plant in China. The full-length cDNA is 3 223 bp long, containing an open reading frame (ORF) of 2 790 bp. Protein sequence comparison showed that the pupative amino acid sequence of this ORF shares high similarity to trehalose-6-phosphate synthases of mode speciese, especially at the conserved sites of catalytic activity centers. Yeast functinal complementation test showed that trehalose-6-phosphate synthase mutant (tps1△), transformed by the ORF of SpTPS1 gene, can restore growth on the mediam supplemented with glucose as a sole carbon source. This result indicated that SpTPS1 of S. pulvinata encodes for an activative protein and is hopeful to be applied in transgeneic improvement of abiotic stress tolerance in plant.
LIN Jing, FU Feng-Ling, JIANG Wei, MAO Yu, YONG Ta-Meng, LI Wan-Chen . Cloning and functional analysis of trehalose -6-phosphate synthase gene from Selaginella pulvinata[J]. Hereditas(Beijing), 2010 , 32(5) : 498 -504 . DOI: 10.3724/SP.J.1005.2010.00498
[1] 王关林, 方宏筠主编. 植物基因工程(第二版). 北京: 科学出版社, 2002, 61–73.
[2] Colaco C, Kampinga J, Roser B. Amorphous stability and trehalose. Science, 1995, 268(5212): 788.
[3] Crowe JH, Crowe LM, Chapman D. Preservation of mem-branes in anhydrobiotic organisms: the role of trehalose. Science, 1984, 223(4637): 701–703.
[4] Kaasen I, Falkenberg P, Styrvold OB, strom AR. Molecu-lar cloning and physical mapping of the otsB genes, which encode the osmoregulatory trehalose pathway of Es-cherichia coli: evidence that transcription is activated by katF (AppR). J Bactreiol, 1992, 174(3): 889–898.
[5] Bell W, Klaassen P, Ohnacker M, Boller T, Herweijer M, Schoppink P, Van der Zee P, Wiemken A. Characterization of the 56-kDa subunit of yeast trehalose-6-phosphate syn-thase and cloning of its gene reveal its identity with the product of CIF1, a regulator of carbon catabolite inactiva-tion. Eur J Biochem, 1992, 209(3): 951–959.
[6] Zentella R, Mascorro-Gallardo JO, Van Dijck P, Folch-Mallol J, Bonini B, Van Vaeck C, Gaxiola R, Co-varrubias AA, Nieto-Sotelo J, Thevelein JM, Iturriaga G. A Selaginella lepidophylla trehalose-6-phosphate synthase complements growth and stress-tolerance defects in a yeast tps1 mutant. Plant Physiol, 1999, 119(4): 1473–1482.
[7] Serrano R, Villalba JM. Expression and localization of plant membrane proteins in Saccharomyces. Methods Cell Biol, 1995, 50: 481–496.
[8] Christianson TW, Sikorski RS, Dante M, Shero JH, Hieter P. Multifunctional yeast high-copy number shuttle vectors. Gene, 1992, 110(1): 119–122.
[9] Elble R. A simple and efficient procedure for transforma-tion of yeasts. BioTechniques, 1992, 13(1): 18–20.
[10] Gibson RP, Turkenburg JP, Charnock SJ, Lloyd R, Davies GJ. Insights into trehalose synthesis provided by the structure of the retaining glucosyltransferase OtsA. Chem Biol, 2002, 9(12): 1337–1346.
[11] Boume Y, Henrissat B. Glycoside hydrolases and glyco-syltransferases: families and functional modules. Curr Opin Struct Biol, 2001, 11(5): 593–600.
[12] Van Aelst L, Hohmann S, Bulaya B, de Koning W, Sierk-stra L, Neves MJ, Luyten K, Alijo R, Ramos J, Coccetti P. Molecular cloning of a gene involved in glucose sensing in the yeast Saccharomyces cerevissiae. Mol Microbiol, 1993, 8(5): 927–943.
[13] Thevelein JM, Hohmann S. Trehalose synthase: guard to the gate of glycolysis in yeast? Trends Biochem Sci, 1995, 20(1): 3–10.
[14] Blazquez MA, Lagunas R, Gancedo C, Gancedo IM. Tre-halose-6-phosphate, a new regulator of yeast glycolysis that inhibits hexokinases. FEBS Lett, 1993, 329(1–2): 51–54.
[15] Blazquez MA, Santos E, Flores CL, Martinez Zapater JM, Salinas J, Gancedo C. Isolation and characterization of the Arobidopsis TPS1 gene. Plant J, 1998, 13(5): 685–689.
[16] Tarczynski MC, Jensen RG, Bohnert HJ. Expression of a bacterial mtlD gene in transgenic tobacco leads to produc-tion and accumulation of mannitol. Proc Natl Acad Sci USA, 1992, 89(7): 2600–2604.
[17] Holmstrom KO, Mantyla E, Welin B, Mandal A, Palva ET, Tunnela OE, Londesborough J. Drought tolerance in to-bacco. Nature, 1996, 379(6567): 683–684.
[18] Goddijn OJ, Verwoerd TC, Voogd E, Krutwagen RW, de Graaf PT, van Dun K, Poels J, Ponstein AS, Damm B, Pen J. Inhibition of trehalose activity enhances trehalose accumulation in trans-genic plants. Plant Physiol, 1997, 113(1): 181–190.
[19] Romero C, Belles JM, Vaya JL, Serrano R, Culianez-Macia FA. Expression of the yeast trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiotropic phenotypes include drought tolerance. Planta, 1997, 201(3): 293–297.
[20] Pilon-Smits E
/
| 〈 |
|
〉 |
www.chinagene.cn
备案号：京ICP备09063187号
