title: Molecular characterization and expression analysis of a GTP-binding protein (MiRab5) in Mangifera indica.
abstract: The Rab family, the largest branch of Ras small GTPases, plays a crucial role in the vesicular transport in plants. The members of Rab family act as molecular switches that regulate the fusion of vesicles with target membranes through conformational changes. However, little is known about the Rab5 gene involved in fruit ripening and stress response. In this study, the MiRab5 gene was isolated from stress-induced Mangifera indica. The full-length cDNA sequence was 984bp and contained an open reading frame of 600bp, which encoded a 200 amino acid protein with a molecular weight of 21.83kDa and a theoretical isoelectric point of 6.99. The deduced amino acid sequence exhibited high homology with tomato (91% similarity) and contains all five characteristic Rab motifs. Real-time quantitative RT-PCR analysis demonstrated that MiRab5 was ubiquitously expressed in various mango tree tissues at different levels. The expression of MiRab5 was up-regulated during later stages of fruit ripening. Moreover, MiRab5 was generally up-regulated in response to various abiotic stresses (cold, salinity, and PEG treatments). Recombinant MiRab5 protein was successfully expressed and purified. SDS-PAGE and western blot analysis indicated that the expressed protein was recognized by the anti-6-His antibody. These results provide insights into the role of the MiRab5 gene family in fruit ripening and stress responses in the mango plant.
authors.affiliation: College of Agriculture, Guangxi University, Nanning 530004, PR China.
authors.forename: Zhao-liang
authors.initials: ZL
authors.lastname: Liu
authors.affiliation: College of Agriculture, Guangxi University, Nanning 530004, PR China.
authors.forename: Cong
authors.initials: C
authors.lastname: Luo
authors.affiliation: College of Agriculture, Guangxi University, Nanning 530004, PR China.
authors.forename: Long
authors.initials: L
authors.lastname: Dong
authors.affiliation: College of Agriculture, Guangxi University, Nanning 530004, PR China.
authors.forename: Can
authors.initials: C
authors.lastname: Van Toan
authors.affiliation: College of Agriculture, Guangxi University, Nanning 530004, PR China.
authors.forename: Peng-xiao
authors.initials: PX
authors.lastname: Wei
authors.affiliation: College of Agriculture, Guangxi University, Nanning 530004, PR China; Guangxi Crop Genetic Improvement and Biotechnology Laboratory, Nanning 530007, PR China. Electronic address: honest66222@163.com.
authors.forename: Xin-hua
authors.initials: XH
authors.lastname: He
mesh_terms: D000595:Amino Acid Sequence; D058639:Cold-Shock Response; D005638:Fruit / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant; D031022:Mangifera / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D055049:Salt Tolerance; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D020696:rab5 GTP-Binding Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D010940:Plant Proteins; D020696:rab5 GTP-Binding Proteins
keywords: Fruit ripening; Mangifera indica; MiRab5 gene; Prokaryotic expression; Stress