The application of expression analysis in elucidating the eukaryotic elongation factor one alpha gene family in Arabidopsis thaliana

Ransom-Hodgkins, Wendy D.

doi:10.1007/s00438-008-0418-2

Cited by 27 publications

(13 citation statements)

References 55 publications

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“…Our previous study showed that one of the eEF1A genes in cassava (MeEF1A1) was expressed in early stages of plant development and also induced by wounding [43]. Recent study in A.thaliana using microarray technique showed that eEF1A gene family was expressed in all tissues but it was also indicated that each eEF1A genes had a unique expression pattern regulated differently by a variety of stimuli [44].…”

Section: Introductionmentioning

confidence: 99%

Isolation and Characterization of Three Cassava Elongation Factor 1 Alpha (MeEF1A) Promoters

2014

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In plant genetic engineering, the identification of gene promoters leading to particular expression patterns is crucial for the development of new genetically modified plant generations. This research was conducted in order to isolate and characterize several new promoters from cassava (Manihot esculenta Crantz) elongation factor 1 alpha (EF1A) gene family. Three promoters MeEF1A3, MeEF1A4 and MeEF1A5 were successfully isolated. Sequence analyses showed that all of the promoters contain three conserved putative cis-acting elements which are located upstream of the transcription start site. These elements are included a TEF1, a TELO and TATA boxes. In addition, all of the promoters also have the 5′UTR intron but with a different lengths. These promoters were constructed translationally with gusA reporter gene (promoter::gusA fusion) in pBI-121 binary vector to build a new binary vector using Overlap Extension PCR Cloning (OEPC) technique. Transient expression assay that was done by using agroinfiltration method was used to show functionality of these promoters. Qualitative and quantitative analysis from GUS assay showed that these promoters were functional and conferred a specific activity in tobacco seedlings (Nicotiana tabacum), tomato fruits (Solanum lycopersicum) and banana fruits (Musa acuminata). We hypothesized that MeEF1A6 could be categorized as a constitutive promoter because it was able to drive the gene expression in all transformed tissue described in here and also comparable to CaMV35S. On the other hand, MeEF1A3 drove specific expression in the aerial parts of seedlings such as hypocotyl and cotyledon thus MeEF1A5 drove specific expression in fruit tissue. The results obtained from transient analysis showed that these promoters had a distinct activity although they came from same gene family. The DNA sequences identified here are new promoters potentially use for genetic engineering in cassava or other plants.

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Section: Introductionmentioning

confidence: 99%

Isolation and Characterization of Three Cassava Elongation Factor 1 Alpha (MeEF1A) Promoters

2014

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“…8). This protein is a member of the family of small G-proteins and serves a multitude of functions (79) including elongation of protein translation (80), regulation of the cytoskeleton (81, 82), and signaling (83–85). It has been previously shown to be methylated (86, 87) and acetylated (88).…”

Section: Resultsmentioning

confidence: 99%

Comprehensive Cell-specific Protein Analysis in Early and Late Pollen Development from Diploid Microsporocytes to Pollen Tube Growth

Ischebeck

Valledor

Lyon

et al. 2014

Molecular & Cellular Proteomics

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Pollen development in angiosperms is one of the most important processes controlling plant reproduction and thus productivity. At the same time, pollen development is highly sensitive to environmental fluctuations, including temperature, drought, and nutrition. Therefore, pollen biology is a major focus in applied studies and breeding approaches for improving plant productivity in a globally changing climate. The most accessible developmental stages of pollen are the mature pollen and the pollen tubes, and these are thus most frequently analyzed. To reveal a complete quantitative proteome map, we additionally addressed the very early stages, analyzing eight stages of tobacco pollen development: diploid microsporocytes, meiosis, tetrads, microspores, polarized microspores, bipolar pollen, desiccated pollen, and pollen tubes. A protocol for the isolation of the early stages was established. Proteins were extracted and analyzed by means of a new gel LC-MS fractionation protocol. In total, 3817 protein groups were identified. Quantitative analysis was performed based on peptide count. Exceedingly stage-specific differential protein regulation was observed during the conversion from the sporophytic to the gametophytic proteome. A map of highly specialized functionality for the different stages could be revealed from the metabolic activity and pronounced differentiation of proteasomal and ribosomal protein complex composition up to protective mechanisms such as high levels of heat shock proteins in the very early stages of development.

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“…EF-1α is reported to have many other functions, including cell growth, motility, protein turnover, and signal transduction (Ridgley et al 1996;Ransom-Hodgkins 2009). In addition, EF-1α can bind actin filaments and microtubules and regulate their assembly and cross-linking (Yang et al 1990;Doyle et al 2011).…”

Section: Discussionmentioning

confidence: 99%

Identification of Eimeria acervulina conoid antigen using chicken monoclonal antibody

et al. 2016

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In the poultry industry, Eimeria spp. is one of the important pathogens which cause significant economic losses. We have previously generated a chicken monoclonal antibody (mAb), 6D-12-G10, with specificity for an antigen located in the apical cytoskeleton of Eimeria acervulina and with cross-reactive among Apicomplexan parasites, including other Eimeria spp., Toxoplasma, Neospora, and Cryptosporidium spp. Furthermore, the protein of Cryptosporidium parvum recognized by the 6D-12-G10 has been identified as elongation factor-1α (EF-1α). In the present study, to identify the target molecule of E. acervulina by the mAb, we performed two-dimensional Western blotting analysis. Finally, we found two positive molecules which are identified as EF-1α and a related protein. Our previous finding using C. parvum and the results in this study suggest that EF-1α could be associated with the invasion facilitated by the cytoskeleton at the apical region of zoites.

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The application of expression analysis in elucidating the eukaryotic elongation factor one alpha gene family in Arabidopsis thaliana

Cited by 27 publications

References 55 publications

Isolation and Characterization of Three Cassava Elongation Factor 1 Alpha (MeEF1A) Promoters

Isolation and Characterization of Three Cassava Elongation Factor 1 Alpha (MeEF1A) Promoters

Comprehensive Cell-specific Protein Analysis in Early and Late Pollen Development from Diploid Microsporocytes to Pollen Tube Growth

Identification of Eimeria acervulina conoid antigen using chicken monoclonal antibody

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