Progress 10/01/02 to 09/30/05
Outputs
Expression of three foreign proteins, namely, beta-glucuronidase (GUS), yeast invertase, and human granulocyte macrophage colony-stimulating factor (GM-CSF) in transgenic sugarcane has been achieved. Accumulation of GM-CSF protein reached 0.02% of total soluble protein in the best transgenic line. The sugarcane-derived GM-CSF shows similar activity as commercially available GM-CSF, toward supporting proliferation of human bone marrow cells (TF-1) which require GM-CSF for cell division. We have also developed an improved fluorescent protein tag useful for reporting gene expression and for protein purification. This tag was shown to improve and accelerate recombinant protein purification from plant extracts. In addition, we have developed a simple three-step purification method for GFP fusion proteins produced in transgenic plants with the intent of maximizing purity and yield under gentle conditions so as to maintain the integrity of the fusion partner.
Impacts
Recombinant protein products represent a multi-billion dollar business worldwide. Transgenic plants are an attractive system for producing recombinant proteins through molecular farming. This research has addressed critical challenges associated with molecular farming, especially on developing secure expression hosts and technologies that improve product recovery. Achieving effective molecular farming could bring about enormous economical impact while reviving modern agricultural development.
Publications
- Wang, M.L., Goldstein, C., Su, W.W., Moore, P.H. and Albert, H.H. 2005. Production of biologically active GM-CSF in sugarcane: a secure biofactory. Transgenic Research. 14:167-178.
- Su, W.W. 2005. Fluorescent proteins as tools to aid protein production. Microbial Cell Factories. 4:12.
- Paramban, R.I. 2003. Engineering GFP as a dual functional tag. MS thesis, University of Hawaii at Manoa, Honolulu, Hawaii.
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Progress 10/01/03 to 09/30/04
Outputs
Results on the expression of human granulocyte macrophage colony-stimulating factor (GM-CSF) in transgenic sugarcane have been published in a manuscript accepted by Transgenic Research for publication. Results on an improved fluorescent protein tag useful for reporting gene expression and for protein purification have been published in Biotechnology & Bioengineering. We have also developed an effective strategy for purifying GFP-tagged recombinant proteins from plant extracts.
Impacts
Recombinant protein products represent a multi-billion dollar business worldwide. Transgenic plants are an attractive system for producing recombinant proteins through molecular farming. This research has addressed critical challenges associated with molecular farming, especially on developing secure expression hosts and technologies that improve product recovery. Achieving effective molecular farming could bring about enormous economical impact while reviving modern agricultural development.
Publications
- Parambam, R.I., Bugos, R. and Su, W.W. 2004. Engineering green fluorescent protein as a dual functional tag. Biotechnol. Bioeng. 86(6):687-97.
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Progress 10/01/02 to 09/30/03
Outputs
We have achieved expression of three foreign proteins, namely, beta-glucuronidase (GUS), yeast invertase, and human granulocyte macrophage colony-stimulating factor (GM-CSF) in transgenic sugarcane. Expression of these proteins in sugarcane was confirmed by western blot analysis. The expression however was found to be unstable. We have also developed an improved fluorescent protein tag useful for reporting gene expression and for protein purification. This tag was shown to improve and accelerate recombinant protein purification from plant extracts.
Impacts
Plant molecular farming, i.e., growing and harvesting genetically altered plants for the production of therapeutics, vaccines, diagnostics, industrial enzymes or biopolymers offers tremendous opportunities for value-added and diversified agriculture. Harvesting and purifying recombinant proteins from transgenic plants remains a major bottleneck in achieving economical plant molecular farming. Researchers at the University of Hawaii have developed a novel fusion protein tag, when fused to a protein of interest; the protein can be easily monitored and purified from plant extracts, and hence substantially reduces the production costs associated with plant molecular farming.
Publications
- Su, W.W. and Arias, R. 2003. Continuous plant cell perfusion culture: bioreactor characterization and secreted enzyme production. J. Biosci. Bioeng. 95(1):13-20.
- Su, W.W., Guan, P.Z. and Bugos, R.C. 2003. High level of secretion of green fluorescent protein in transgenic tobacco cell culture: characterization and sensing. Biotechnol. Bioeng. (In press).
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Progress 10/01/01 to 09/30/02
Outputs
No progress to report. This project was initiated on October 1, 2002.
Impacts
(N/A)
Publications
- No publications reported this period
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