Expression of chalcone synthase and chalcone isomerase proteins in Arabidopsis seedlings.
Antibodies have been developed against the first two enzymes of flavonoid biosynthesis in Arabidopsis thaliana. Chalcone synthase (CHS) and chalcone isomerase (CHI) were overexpressed and purified from Escherichia coli as fusion proteins with glutathione S-transferase from Schistosoma japonicum. The recombinant proteins were then used to immunize chickens and the resulting IgY fraction was purified from egg yolks. Immunoblots of crude protein extracts from Arabidopsis seedlings carrying wild-type and null alleles for CHS and CHI showed that the resulting antibody preparations provide useful tools for characterizing expression of the flavonoid pathway at the protein level. An initial analysis of expression patterns in seedlings shows that CHS and CHI proteins are present at high levels during a brief period of early seedling germination that just precedes the transient accumulation of flavonoid end-products.
Cain
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Department of Biology, Virginia Polytechnic Institute and State University, Blacksburg 24061, USA.
Saslowsky
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Walker
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Shirley
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D000217:Acyltransferases / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000906:Antibodies / Q000737:chemistry / Q000235:genetics; D017360:Arabidopsis; D003001:Cloning, Molecular; D003429:Cross Reactions; D005982:Glutathione Transferase / Q000235:genetics / Q000276:immunology; D015151:Immunoblotting; D019753:Intramolecular Lyases / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology
D000906:Antibodies; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D000217:Acyltransferases; C020141:flavanone synthetase; D005982:Glutathione Transferase; D019753:Intramolecular Lyases; C019865:chalcone isomerase