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KMID : 0903519900330040353
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
1990 Volume.33 No. 4 p.353 ~ p.353
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Cloning and expression of the soybean Bowman - Birk trypsin isoinhibitor D - Ⅱ in E . coli
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Baek Jong-Min
Kim Su-Il
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Abstract
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A cDNA clone for the soybean trypsin inhibitor (Bowman-Birk type) was isolated from a soybean λgtll cDNA lihrary by plaque hybridization with oligonucleotide probe. A clone with 460 base pairs insert was isolated and confirmed by DNA sequencing. The cDNA clone B2 was consisted of 459 nucleotides-250b.p. of open reading frame, 92b.p. of 5´-nontranslating region and 117b.p. of 3´-nontranslating region. Compared the predicted amino acid sequence, it turned out that the protein encoded by the cDNA clone B2 was the Bowman-Birk type proteinase isoinhibitor D- II .
Expression plasmids were consisted in which the B2 insert was inserted into Sma I site of pUC18 polycloning region by blunt-end ligation. The plasmid expressed chimeric polypetide was identified by SDS-gel electrophoresis and Western blot analysis.
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