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Isolation and expression of a eukaryotic translation initiation factor 5A gene from sugarcane(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2015 06
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Isolation and expression of a eukaryotic translation initiation factor 5A gene from sugarcane
QUE Wancai HUANG Ning LIU Feng XIAO Xinhuan LING Hui ZHANG Yuye SU Weihua SU Yachun WU Qibin QUE Youxiong
1Department of Pharmacy, Fujian Medical University Union Hospital, Fuzhou 350001, China 2Key Laboratory of Sugarcane Biology and Genetic Breeding (Fujian), Ministry of Agriculture, Fujian Agriculture and Forestry University/Sugarcane Research & Development Center, China Agricultural Technology System, Fuzhou 350002, China
sugarcane eIF5A Sporisorium scitamineum bioinformatics Real-time Quantitative PCR
Q78 : S566.103.4

Eukaryotic translation initiation factor 5A (eIF5A) is ubiquitous in animals, plants and fungi. To reveal the responses of sugarcane eIF5A gene to biotic stresses, the present study first obtained an EST sequence with 93% homologs to Zea mays eIF5A (GenBank Accession Number:EU958725.1) from a suppression subtractive hybridization (SSH) library of sugarcane challenged by Sporisorium scitamineum. The EST sequence was then used as the probe for in silico cloning to obtain a putative cDNA sequence of sugarcane eIF5A gene. This gene was validated by reverse transcription-PCR (RT-PCR) amplification sequencing, which showed that the sequence cloned by in silico cloning was in accordance with the sequence amplified by RT-PCR method, and was named as SceIF5A (GenBank Accession Number: KJ577595). Bioinformatics analysis indicated that SceIF5A, with a length of 1 174 bp, containing a 483 bp open reading frame (ORF) encoding 160 amino acids protein, was a stable acidic cytoplasm protein with a weight of 17 453.6 Da containing 12 highly conserved amino acid sequences. Real-time quantitative PCR analysis showed that the expression of SceIF5A was up-regulated under smut fungus infection and the treatments of SA, MeJA and ABA. The results suggested that SceIF5A in sugarcane is most probably involved in response to sugarcane smut fungus infection and to hormone related bioprocess signaling. The results of this study regarding the structure and function of SceIF5A should lay the foundation for further research in the elaboration of functions of this gene during the interaction between sugarcane and S. scitamineum.


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