Location: Crop Improvement and Genetics Research
2012 Annual Report
2) Systematically reconstruct the latex composition from synthetic and Hevea, guayule, and Russian dandelion natural latex by protein add-back compositions.
3) Fully characterize the nanostructure and properties of resulting materials.
In FY2012 collaborative research established that addition of amino acids to guayule latex improve the thermo-oxidative stability, regardless of the R functional group. In other words, the peptide bond nature, more specifically, the N from the peptide bond, may participate in rubber stabilization. Separately, expression and purification of Hevea REF (a 14 KDa. protein) and SRPP proteins were carried out using a pGEX-6P-1 vector transferred to E. coli competent cells (Rossetta cells), incubation time was optimized, and the protein purified according to the purification protocols of GST fusion proteins. Unfortunately the protein concentration was lower than desired (about 1.1mg per 500 ml culture). Using a High Cell Density method did not show significant increase in protein expression for both REF and SRPP. It is possible that REF and SRPP in high concentrations have toxic effects on bacterial cell. Our next step will be to try a baculovirus system for cloning and high-level expression of recombinant proteins, using Gateway Technology. In this case the recombinant baculovirus will be used to directly transfect insect cells, which may be less sensitive to high levels of recombinant protein.