Cloning and expression analysis ofeIF-5Agene inApocynum venetum

Abstract

To understand the effects of the eIF-5A gene in response to abiotic stress, the 909 bp full-length cDNA of eIF-5A, including a 480 bp open reading frame encoding 159 amino-acid (aa) residues, was isolated from the leaf of herbal plant (Apocynum venetum) by rapid-amplification of cDNA ends. The deduced molecular weight of the encoding protein was 17.48 kDa with a theoretical pI of 5.61 and predicted no signal peptide. Real-time polymerase chain reaction analysis revealed that AveIF-5A gene expression was induced by cold, salt and drought stress. To determine the biological function of this gene, recombinant plasmids expressing AveIF-5A and AveIF-5A1 (86À156 aa truncated polypeptide) were used to transform Escherichia coli. The analysis of the growth curves of recombinant E. coli revealed that AveIF-5A and AveIF-5A1 improved the resistance of transformed E. coli to low-temperature, drought and salt stress. These results could provide experimental evidence of the function of the AveIF-5A gene as a valuable gene resource in plant resistance-breeding.

Hypusination, a Metabolic Posttranslational Modification of eIF5A in Plants during Development and Environmental Stress Responses

“…In the case of Hordeum vulgare, it was found that eIF5A-1 isoforms are more abundant in H. vulgare, inducing apoptosis, while the salt-tolerant H. maritimum contained more eIF5A-2 isoforms responsible for cell division [70]. Wang et al (2016) [51] conducted a study on Apocynum venetum eIF5A1, which is a salt-tolerant halophyte herb plant. These studies suggest a cultivar-and a species-dependent mechanism of hypusination and eIF5A isoforms in plants.…”

“…In addition, if eIF5A does in fact confer stress tolerance in plants, the physiological changes mediated by eIF5A deserve further study. Studying AveIF5A from Apocynum venetum , a halophyte shrub, showed that its expression profile was different for diverse stress conditions, such as cold, salt or drought stress [ 51 ]. These results indicated that eIF5A could play an important role in some abiotic stress conditions, and the timing of stress could imply the success of eIF5A-promoted tolerance processes.…”

Engineering the bacterial endophyte Burkholderia pyrrocinia JK-SH007 for the control of lepidoptera larvae by introducing the cry218 genes of Bacillus thuringiensis

“…Protein samples were separated on a 4%-12% Tris-glycine gel, and then transferred onto a nitrocellulose membrane. The membrane was soaked in 10 mL TBS-T (20 mmol/L Tris-HCl, 150 mmol/L sodium chloride, 0.05% Tween 20) with 10% (w/v) milk for 1 h at room temperature [14]. Western blot analysis was conducted using primary antibody (Mouse Monoclonal Anti-6 £ His IgG antibody, 1:3,000) and a secondary antibody IgG-HRP (horseradish peroxidase-conjugated goat anti-Mouse IgG antibody, 1:5000).…”