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Molecular cloning and characterization of a cassava translationally controlled tumor protein gene potentially related to salt stress response

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Abstract

Cassava (Manihot esculenta Crantz) is one of the most important tropical crops showing tolerance to abiotic stress and adaptations to a wide range of environmental conditions. Here, we aimed to isolate and characterize the full-length cDNA and genomic sequences of a cassava translationally controlled tumor protein gene (MeTCTP), and evaluate its potential role in response to salt stress. The MeTCTP full-length cDNA sequence encodes for a deduced protein with 168 amino acid residues, with theoretical isoelectric point and molecular weight of 4.53 and 19 kDa, respectively, containing two putative signatures of TCTP family and one site for myristoylation. The MeTCTP genomic sequence includes four introns and five exons within a 1,643 bp coding region, and a 264 bp partial promoter sequence containing several putative cis-acting regulatory elements, among them, two putative GT-1 motifs, which may be related to response to sodium chloride (NaCl) and pathogen infection. Semi-quantitative RT-PCR assays showed that MeTCTP transcripts were higher in roots than leaves, and were significantly increased in detached leaves treated with NaCl. Furthermore, the recombinant MeTCTP conferred a protective function against salt stress in bacterial cells. We report for the first time the molecular cloning and characterization of a cassava TCTP with potential role in salt-stress response. Since salinity is one the most important abiotic factors affecting the production of crops worldwide, the MeTCTP gene could be a candidate gene for generation of salt tolerant crops.

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Acknowledgments

This work was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Pará (FAPESPA), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and Universidade Federal do Pará (UFPA), Brazil.

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  1. Instituto de Ciências Biológicas, Universidade Federal do Pará, Guamá, Belém, PA, 66075-110, Brazil

    Ailton Borges Santa Brígida, Sávio Pinho dos Reis, Carinne de Nazaré Monteiro Costa, Cristina Michiko Yokoyama Cardoso, Aline Medeiros Lima & Cláudia Regina Batista de Souza

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  1. Ailton Borges Santa Brígida
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  2. Sávio Pinho dos Reis
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Correspondence to Cláudia Regina Batista de Souza.

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Supplementary Table 1 Primers sequences used in the isolation and characterization of MeTCTP. (DOC 33 kb)

11033_2014_3028_MOESM2_ESM.doc

Supplementary Fig. 1 The full-length cDNA sequence and deduced amino acid sequence of MeTCTP. Start and stop codons are boxed. The potential signal of polyadenilation is underlined. This nucleotide sequence was registered in the NCBI GenBank under accession no. JX855123. (DOC 39 kb)

11033_2014_3028_MOESM3_ESM.doc

Supplementary Fig. 2 Genomic sequence of MeTCTP gene containing coding region and partial promoter sequences. Putative cis-acting regulatory elements are highlighted. Deduced amino acid sequence is in bold capital letters. Introns are in lower case with conserved dinucleotides GT and AG underlined. This nucleotide sequence was registered on GenBank under accession no. JX855122. (DOC 37 kb)

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Santa Brígida, A.B., dos Reis, S.P., de Nazaré Monteiro Costa, C. et al. Molecular cloning and characterization of a cassava translationally controlled tumor protein gene potentially related to salt stress response. Mol Biol Rep 41, 1787–1797 (2014). https://doi.org/10.1007/s11033-014-3028-6

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