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Sequence and expression of the mRNA encoding the chymotrypsin inhibitor in winged bean (Psophocarpus tetragonolobus (L.) DC.) |
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| Authors: | Surin Peyachoknagul Tamotsu Matsui Hiroshi Shibata Saburo Hara Tokuji Ikenaka Yoshimi Okada Takeshi Ohno |
| Institution: | (1) Department of Agricultural Chemistry, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 183 Fuchu, Tokyo, Japan;(2) Department of Chemistry, Osaka University College of Science, 560 Toyonaka, Osaka, Japan;(3) Department of Biophysics and Biochemistry, Faculty of Science, The University of Tokyo, 113 Tokyo, Japan;(4) Present address: Department of Agricultural Chemistry, Faculty of Agriculture, Hokkaido University, 060 Sapporo, Japan |
| Abstract: | The accumulation of the Kunitz-type chymotrypsin inhibitor WCI-3 in winged bean seeds is controlled developmentally. In vitro translation experiments showed that the WCI-3 mRNA was present in 35- and 40-day-old immature seeds after flowering. The size of the in vitro translation product is about 2 000 Da larger than that of the mature WCI-3 protein. The WCI-3 cDNA clones were isolated from a  gtll cDNA library of 35-day-old immature seeds by immunoscreening. A nearly full-length cDNA clone was obtained containing an open reading frame of 207 amino acid residues. The deduced sequence of the 183 carboxy terminal amino acids coincides precisely with the amino acid sequence determined for purified WCI-3. The amino terminal extension of 24 residues has the characteristics of a signal peptide. Northern hybridization analysis of total poly(A)+ RNA showed that the WCI-3 mRNA is approximately 900 nucleotides long and accumulates in 35- and 40-day-old but not in 30-day-old immature seeds. |
| Keywords: | cDNA sequence expression library in vitro translation Kunitz-type proteinase inhibitor mRNA accumulation in vivo signal peptide |
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