Abstract
A genomic gene of tritin, a ribosome-inactivating protein (RIP) from Triticum aestivum, was cloned using a barley RIP gene as a probe. The 5′-non-coding region has potential TATA boxes and three sequences homologous to the binding sequence of the transcriptional activator protein Opaque-2 which activates maize RIP gene expression. The cloned DNA encoded tritin consits of 275 amino acids with no secretion signal sequence. The coding region of tritin was expressed in Escherichia coli using lac promoter and yielded a protein similar to the native one, as determined by SDS-polyacrulamide gel electrophoresis and immunological analysis.
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Abbreviations
- MAP:
-
Mirabilis antiviral protein
- RIP:
-
ribosome-inactivating protein
- SDS-PAGE:
-
sodium dodecyl sulfate polyacrylamide gel electrophoresis
- IC50 :
-
median of inhibitory concentration of L-[35S] methionine incorporation
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Habuka, N., Kataoka, J., Miyano, M. et al. Nucleotide sequence of a genomic gene encoding tritin, a ribosome-inactivating protein from Triticum aestivum . Plant Mol Biol 22, 171–176 (1993). https://doi.org/10.1007/BF00039007
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DOI: https://doi.org/10.1007/BF00039007