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Molecular Genetics and Genomics

, Volume 281, Issue 4, pp 391–405 | Cite as

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

  • Wendy Danielle Ransom-Hodgkins
Original Paper

Abstract

Eukaryotic elongation factor one alpha (eEF1A) encoding genes are part of the large GTP binding protein family. The eEF1A family is important for protein synthesis and actin filament and bundle formation. In this study, the expression of four eEF1A genes in Arabidopsis thaliana is reported. Microarray analyses of the gene family showed high expression levels in germinating seeds, embryos, and shoot and root meristems. Quantitative real time RT-PCR was used to determine individual eEF1A gene expression. Unlike animals, in Arabidopsis tissues all four eEF1A genes were expressed in all tissues sampled. However, the abundance of each transcript varied spatially. Knocking out expression of one eEF1A gene produced seedlings with stunted roots and a subsequent change in expression of the other three eEF1A genes. The varying abundance of each gene in different tissues may indicate different concentration requirements for each message product. These results will be very useful for elucidating the role of each gene in growth, development, and stress responses of the plant.

Keywords

eEF1A Quantitative real time RT-PCR Arabidopsis thaliana Development Microarray Roots 

Abbreviations

eEF1A

Eukaryotic elongation factor one alpha

ACT,2,8

Actin2 and Actin8 genes

UBQ10

Polyubiquitin 10 gene

Q real time RT-PCR

Quantitative real time reverse transcription polymerase chain reaction

GUS

β-glucuronidase

A1

At1g07920

A2

At1g07930

A3

At1g07940

A4

At5g60390

ORF

Open reading frame

GFP

Green fluorescent protein

CDS

Coding sequence

UTR

Untranslated region

NPA

1-N-naphthylphthalamic acid

ABA

Abscisic acid

BL

Brassinolide

GA3

Gibberellin

AVG

Aminoethoxyvinylglycine

MJ

Methyl jasmonate

Notes

Acknowledgments

I would like to thank Dr. Todd Barkman for sequencing the PCR products; and Dr. Charles Ide and Dr. Anna Jelaso for use of the Primer Express Software and assistance in the cDNA microarray analysis. This work was supported by The College of Arts and Sciences, Western Michigan University to W.R.H.

Supplementary material

438_2008_418_MOESM1_ESM.tif (739 kb)
Supplemental Figure 1. Quantitative real time RT-PCR products.The PCR products from reactions using the eEF1A family A1, A2, A3, A4; ACT2,8 or UBQ10primers were analyzed using a DNA500 Lab on a Chip kit in a 2100 Bioanalyzer (AgilentTechnologies). The PCR products are shown. Each PCR product was cloned into pCR4 andsequenced to verify amplification of the correct gene. The sizes of the PCR products are: A1 200bp; A2 186bp; A3 194bp; A4 178bp; UBQ10 74bp; ACT2,8 73bp; and eEF1A Family 75bp(TIFF 739 kb)
438_2008_418_MOESM2_ESM.tif (49.2 mb)
Supplemental Figure 2. Expression of UBQ10 and ACT2,8. Quantitative real time RT-PCR was performed on RNA extracted from developing tissue. Refer to Figure 5B for measurement details. The experiment was repeated twice with triplicate samples for each treatment. The standard deviation is reported.(TIFF 50337 kb)
438_2008_418_MOESM3_ESM.doc (77 kb)
Supplementary Table 1 (DOC 77 kb)
438_2008_418_MOESM4_ESM.doc (30 kb)
Supplementary Table 2 (DOC 30 kb)

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Department of Biological SciencesWestern Michigan UniversityKalamazooUSA

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