Abstract
S-Gene-associated glycoproteins (S-glycoproteins) from styles of Nicotiana alata, identified by non-equilibrium two-dimensional electrophoresis, were purified by cation exchange fast protein liquid chromatography with yields of 0.5 to 8 micrograms of protein per style, depending on the S-genotype of the plant. The method relies on the highly basic nature of the S-glycoproteins. The elution profiles of the different S-glycoproteins from the fast protein liquid chromatography column were characteristic of each S-glycoprotein, and could be used to establish the S-genotype of plants in outbreeding populations. In all cases, the S-genotype predicted from the style protein profile corresponded to that predicted from DNA gel blot analysis using S-allele-specific DNA probes and to that established by conventional breeding tests. Amino-terminal sequences of five purified S-glycoproteins showed a high degree of homology with the previously published sequences of N. alata and Lycopersicon esculentum S-glycoproteins.
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- Anderson M. A., McFadden G. I., Bernatzky R., Atkinson A., Orpin T., Dedman H., Tregear G., Fernley R., Clarke A. E. Sequence variability of three alleles of the self-incompatibility gene of Nicotiana alata. Plant Cell. 1989 May;1(5):483–491. doi: 10.1105/tpc.1.5.483. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Jahnen W., Lush W. M., Clarke A. E. Inhibition of in Vitro Pollen Tube Growth by Isolated S-Glycoproteins of Nicotiana alata. Plant Cell. 1989 May;1(5):501–510. doi: 10.1105/tpc.1.5.501. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Nielsen B. L., Brown L. R. The basis for colored silver-protein complex formation in stained polyacrylamide gels. Anal Biochem. 1984 Sep;141(2):311–315. doi: 10.1016/0003-2697(84)90047-2. [DOI] [PubMed] [Google Scholar]
- O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
- Simpson R. J., Moritz R. L., Lloyd C. J., Fabri L. J., Nice E. C., Rubira M. R., Burgess A. W. Primary structure of ovine pituitary basic fibroblast growth factor. FEBS Lett. 1987 Nov 16;224(1):128–132. doi: 10.1016/0014-5793(87)80435-0. [DOI] [PubMed] [Google Scholar]
- des Francs C. C., Thiellement H., de Vienne D. Analysis of Leaf Proteins by Two-Dimensional Gel Electrophoresis: Protease Action as Exemplified by Ribulose Bisphosphate Carboxylase/ Oxygenase Degradation and Procedure to Avoid Proteolysis during Extraction. Plant Physiol. 1985 May;78(1):178–182. doi: 10.1104/pp.78.1.178. [DOI] [PMC free article] [PubMed] [Google Scholar]