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Molecular characterization of low-temperature inducible genes from mungbean KCI 등재
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/318671
We isolated two low temperature-inducible cDNAs designated as MLT5 and MLT31 from mungbean by subtractive suppression hybridization method. By rapid amplification of cDNA end technique, the full-length cDNAs designated MLT5 and MLT31 were obtained. The full-length cDNA of MLT5 contains an open reading frame of 324 nucleotides coding for 107 amino acids. The full-length cDNA of MLT31 contains an open reading frame of 444 nucleotides in length and capable of specifying a 16.5-kDa protein of 148 amino acids (aa) with an isoelectric point of 7.72. RNA expression of MLT5 was strongly induced by low temperature in the early time and also by wounding, NaCl and ABA. MLT31 mRNA was induced by NaCl and ABA but not by wounding and low temperature stress. MLT5 encodes a protein of unknown function, of which targeting site is predicted to be chloroplast membrane protein. To examine the localization of MLT5, MLT5-GFP was expressed in tobacco cells. It was shown that MLT5-GFP was localized to surface of chloroplast in tobacco cell. To examine the function of MLT31, MLT31 was expressed in Escherichia coli as His-fusion protein. Purified MLT31-His recombinant protein will be tested for ubiquitination activity in vitro. In addition, for the in vivo functional analysis of MLT31, MLT31 will be expressed in yeast ubc9 knock-out mutant. We propose that MLT5 and MLT31 play an important role in signal pathway of abiotic stress in plants.
