A synthetic gene increases TGFβ3 accumulation by 75-fold in tobacco chloroplasts enabling rapid purification and folding into a biologically active molecule
Article first published online: 27 APR 2011
© 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd
Plant Biotechnology Journal
Special Issue: Chloroplast Biotechnology
Volume 9, Issue 5, pages 618–628, June 2011
How to Cite
Gisby, M. F., Mellors, P., Madesis, P., Ellin, M., Laverty, H., O’Kane, S., Ferguson, M. W. J. and Day, A. (2011), A synthetic gene increases TGFβ3 accumulation by 75-fold in tobacco chloroplasts enabling rapid purification and folding into a biologically active molecule. Plant Biotechnology Journal, 9: 618–628. doi: 10.1111/j.1467-7652.2011.00619.x
- Issue published online: 6 MAY 2011
- Article first published online: 27 APR 2011
- Received 7 November 2010; revised 9 March 2011; accepted 9 March 2011.
Figure S1 Comparison of the native TGFβ3 DNA sequence (TGFβ3N) and the synthetic TGFβ3 (TGFβ3O) sequence containing frequent codons present in tobacco chloroplast genes.
Figure S2 Very young leaves of TGFβ3O plants appeared pale-green compared to young WT leaves but rapidly turned green as the leaves matured.
Figure S3 Western blot analysis of TGFβ3 in young (Y), mature (M) and old (O) leaves of TGFβ3N and TGFβ3O plants against known amounts of TGFβ3 standard (STD).
Figure S4 Protein blot analyses on SDS-PAGE fractionated total leaf protein from three TGFβ3O lines (lines 3E, 8H and 9B) from independent transformation events.
Figure S5 Expression of TGFβ3 from plastid transformation vectors p201 and p202 in Escherichia coli.
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