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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Production of a His-Tagged Canecystatin in Transgenic Sugarcane and Subsequent Purification

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Autor(es):
Ribeiro, Carolina W. [1] ; Soares-Costa, Andrea [1] ; Falco, Maria Cristina [2] ; Chabregas, Sabrina M. [2] ; Ulian, Eugenio C. [2] ; Cotrin, Simone S. [3] ; Carmona, Adriana K. [3] ; Santana, Lucimeire A. [3] ; Oliva, Maria Luiza V. [3] ; Henrique-Silva, Flavio [1]
Número total de Autores: 10
Afiliação do(s) autor(es):
[1] Univ Fed Sao Carlos, Mol Biol Lab, Dept Genet & Evolut, BR-13565905 Sao Carlos, SP - Brazil
[2] Ctr Tecnol Canavieira, BR-13400970 Piracicaba, SP - Brazil
[3] Univ Fed Sao Paulo, Escola Paulista Med, BR-04044020 Sao Paulo - Brazil
Número total de Afiliações: 3
Tipo de documento: Artigo Científico
Fonte: BIOTECHNOLOGY PROGRESS; v. 24, n. 5, p. 1060-1066, SEP-OCT 2008.
Citações Web of Science: 8
Resumo

Transgenic plants have been used widely as expression systems of recombinant proteins in recent years. This process can be an efficient alternative for the large-scale production of proteins. In this work, we Present the establishment of transgenic sugarcane expressing a His-tagged canecystatin under the control of the maize ubiquitin promoter. A number of studies have demonstrated that cystatins, which are natural inhibitors of cysteine proteinases, can be used for protection against insect attacks. A transformed sugarcane plant that presented high levels of (HIS)CaneCPI-1 expression, was selected for the purification of this protein through affinity chromatography in a nickel column. This purified (HIS)CaneCPl-1 was immnunodetected using a polyclonal antibody, which was also able to detect the (HIS)CaneCPI-1 in a crude extract from transgenic plant leaves. Assays of inhibitory activity performed with the purified (HIS)CaneCPI-1 revealed its ability to inhibit the catalytic activity of midgut cysteine proteinase partially purified from the sugarcane weevil Sphenophorus levis and human cathepsin L in nanomolar order. These studies demonstrate that sugarcane is a viable expression system for recombinant protein production. (AU)

Processo FAPESP: 98/14138-2 - Center for Structural Molecular Biotechnology
Beneficiário:Glaucius Oliva
Linha de fomento: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
Processo FAPESP: 05/59833-5 - Desenvolvimento de canas-de-açúcar transgênicas superexpressando cistatinas visando o aumento da resistência a insetos
Beneficiário:Andrea Soares da Costa Fuentes
Linha de fomento: Bolsas no Brasil - Pós-Doutorado