A Nicotiana tabacum cell line was transformed, using particle bombardment, with constructs encoding the Brassica stigma glycoproteins, S-locus glycoprotein(SLG)-6 and S-locus related glycoprotein-1, under the control of a duplicated cauliflower mosaic virus 35S promoter and the nos terminator sequence. Calli and cell suspension lines that express each of the two proteins were recovered. Using specific antibodies, the foreign proteins can be detected in the cells, the cell wall eluate and the culture medium. The characteristic band patterns representing glycoforms (and isoforms) of the transgenically produced proteins on sodium dodecyl sulfate and isoelectric focusing protein gels are very similar to those from Brassica stigmas. Partial purification of SLG from the culture medium is described and the potential of the system for studying the biochemical basis of self-incompatibility is discussed.
Bibliographical noteFunding Information:
We wish to thank Rody Spivey and James Blankenship of the Plant Science Center at Cornell for their invaluable help in plant transformation procedures. We thank Dr. W. Crossby of the National Research Council, Canada, for providing the double-35S promoter. R.P.-T. was supported by a post-doctoral fellowship from the Yad Hanadiv-Rotschield Foundation, Jerusalem and M.E.N. was supported by a CIBA-GEIGY grant.
- Particle bombardment
- Protein production
- S-locus glycoprotein (SLG)
- S-locus related glycoprotein-1 (SLR1)
- Tobacco cell culture