Cloning and Functional Analysis of the MtLEA5B Gene from Medicago truncatula

doi:10.13560/j.cnki.biotech.bull.1985.2018-0095

Abstract

Abstract: Late embryogenesis abundant protein（LEA protein）is hydrophilic one that is involved in the responses to stress in plants. This work is aimed to unravel the protective effect of heterologous expressed LEA protein on host under abiotic stress. A novel gene encoding LEA_5 protein was cloned from Medicago truncatula seedlings,and designated as MtLES5B,and its expression patterns were analyzed by semi-quantitative RT-PCR. Then the heat resistance and solubility of recombinant protein were detected by SDS-PAGE protein electrophoresis. Further,the prokaryotic expression vector was constructed and transferred to Escherichia coli so as to induce MtLEA5B over expression in E. coli. The expression patterns of MtLEA5B gene was regulated by dehydration,low temperature,high salt stress,and ABA induction. There was a band by SDS-PAGE protein electrophoresis,which was larger than the theoretical prediction value and did not decrease in solubility under boiling water condition. The overexpression of MtLEA5B protein in E. coli obviously increased the survival rate of hosts at heat（55℃）and freeze（-20℃）. Moreover,MtLEA5B was an inducible expressed gene,MtLEA5B protein had high hydrophily and heat resistance,overexpression in E. coli may significantly enhance its tolerance to temperature stress.

Key words: Medicago truncatula, late embryogenesis abundant protein, MtLEA

ZHANG Ye-meng, SHEN Ying-fang, WANG Ying-fang, YAO Pin-ya, WANG Hai-qing. Cloning and Functional Analysis of the MtLEA5B Gene from Medicago truncatula[J]. Biotechnology Bulletin, 2018, 34(7): 101-107.

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