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

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

Autor Ribeiro, Carolina Werner Google Scholar
Soares-Costa, Andrea Google Scholar
Falco, Maria Cristina Google Scholar
Chabregas, Sabrina Moutinho Google Scholar
Ulian, Eugenio Cesar Google Scholar
Cotrin, Simone Silva Autor UNIFESP Google Scholar
Carmona, Adriana Karaoglanovic Autor UNIFESP Google Scholar
Santana, Lucimeire Aparecida de Autor UNIFESP Google Scholar
Oliva, Maria Luiza Vilela Autor UNIFESP Google Scholar
Henrique-Silva, Flávio Google Scholar
Instituição Universidade Federal de São Carlos (UFSCar)
Ctr Tecnol Canavieira
Universidade Federal de São Paulo (UNIFESP)
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.
Palavra-chave Saccharum
cystatin
phytocystatin
recombinant protein
plant pathogen
cysteine protease inhibitor
Sphenophorus levis
Idioma Inglês
Financiador Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Centro de Tecnologia Canavieira
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Número do financiamento FAPESP: 98/14138-2
FAPESP: 05/598335
Data de publicação 2008-09-01
Publicado em Biotechnology Progress. Washington: Amer Chemical Soc, v. 24, n. 5, p. 1060-1066, 2008.
ISSN 8756-7938 (Sherpa/Romeo, fator de impacto)
Publicador Amer Chemical Soc
Extensão 1060-1066
Fonte http://dx.doi.org/10.1002/btpr.45
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:000260738900007
Endereço permanente http://repositorio.unifesp.br/handle/11600/30858

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