2000年2月
Cloning and expression of the Momordica charantia trypsin inhibitor II gene in silkworm by using a baculovirus vector
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
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- 巻
- 64
- 号
- 2
- 開始ページ
- 393
- 終了ページ
- 398
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1271/bbb.64.393
- 出版者・発行元
- TAYLOR & FRANCIS LTD
MCTI-II (Momordica charantia trypsin inhibitor II) isolated from bitter gourd (Momordica charantia LINN,) seeds is one of the serine protease inhibitors of the squash family. We cloned cDNA that encodes MCTI-II and constructed an expression system for MCTI-II by using a baculovirus vector. The recombinant baculovirus was inoculated to early fifth-instar larvae of the silkworm (strain: Shunrei x Shougetsu). Four days after infection, the hemolymph of silkworm larvae was collected and the recombinant protein was purified. Two kinds of expressed MCTI-II protein were obtained. An amino acid sequence analysis of the two proteins indicates that both were similar to the authentic inhibitor, except for the addition of a tripeptide derived from the vector at the N-terminus. One of the two inhibitors (MCTI-II A) resulted in a single PTH-amino acid in each Edman degradation cycle, while the other (MCTI-II B) resulted in two PTH-amino acids, suggesting the occurrence of cleavage of the reactive site. The inhibitory activities of MCTI-II expressed toward trypsin are examined in terms of the Ki value, these being 6.4x10(-10) M for MCTI-II A and 5.2x10(-10) M for MCTI-II B.
- リンク情報
- ID情報
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- DOI : 10.1271/bbb.64.393
- ISSN : 0916-8451
- eISSN : 1347-6947
- CiNii Articles ID : 110002679922
- Web of Science ID : WOS:000085778200027