| Full text | |
| Author(s): |
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]
Total Authors: 10
|
| Affiliation: | [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
Total Affiliations: 3
|
| Document type: | Journal article |
| Source: | BIOTECHNOLOGY PROGRESS; v. 24, n. 5, p. 1060-1066, SEP-OCT 2008. |
| Web of Science Citations: | 8 |
| Abstract | |
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) | |
| FAPESP's process: | 98/14138-2 - Center for Structural Molecular Biotechnology |
| Grantee: | Glaucius Oliva |
| Support Opportunities: | Research Grants - Research, Innovation and Dissemination Centers - RIDC |