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Isolamento, caracterização e expressão em sistema bacteriano de um gene que codifica uma proteína transportadora de lipídeos de Piper nigrum
(Universidade Federal do Pará, 2012-05-11) BRITO, Wendell Upton de; SOUZA, Cláudia Regina Batista de; http://lattes.cnpq.br/3633972375952822
Black pepper (Piper nigrum L.) is one of the most widely used spices in the world belonging to the Piperaceae family, which comprises about 1400 species distributed mainly in the American tropics and Southern Asia, where this crop originated. Black pepper was introduced in Brazil in the 17th century and has been a nationally important crop since 1933. The Pará State is the main Brazilian producer of black pepper; however, the spice‘s production has been damaged by the root rot disease caused by Fusarium solani f. sp. piperis. Previous studies have revealed the identification of some cDNA sequences differentially expressed during compatible black pepper - F. solani f. sp. piperis interaction. Among them, a partial cDNA sequence coding for a putative Lipid transfer protein (LTP), which is know to play important roles in plant defense against insects and pathogens. Therefore, the aim of this work was to isolate and characterize the full-length cDNA and genomic sequences of black pepper LTP, named PnLTP. The PnLTP full-length cDNA isolated by RACE assays showed 621 bp with 32 bp and 235 bp non-coding sequences at the 5‘ and 3‘ ends, respectively. This cDNA contains a 354-bp ORF coding for a deduced protein with 117 amino acid residues that showed high identity with LTPs from different crops. Sequence analysis revealed that PnLTP contains a putative peptide signal at amino-terminal and eight cysteine residues predicted to form four disulfide bridges, which could contribute to protein stability. Alignment between genomic and cDNA sequences revealed no introns within PnLTP coding region, that is in accordance with other LTP plant genes. Finally, the mature PnLTP was expressed in bacteria system. Additional experiments will be performed in order to evaluate the ability of recombinant PnLTP in inhibit the F. solani f. sp. piperis growth.