| Project/Area Number |
04670138
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
General medical chemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
YAMAMOTO Kazuo The University of Tokyo, Faculty of Pharmaceutical Sciences, Assistant Professor, 薬学部, 助手 (20174782)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1992: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | lectin / pharge / chimeric protein / sugar specificity / 糖鎖 / 相互作用 / 特異性 / キメラ |
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| Research Abstract |
The carbohydrate-binding domain of Bauhinia purpurea lectin (BPA), was investigated from Asp-N endoproteinase digests of BPA.A peptide which interacted with lactose was purified by means of lactose-Sepharose column chromatography. It consisted of 9 amino acids. Subsequently, we constructed a chimeric lectin gene by using a cDNA coding BPA in which nonapeptide sequence was replaced by the corresponding region of the mannose-binding Lens culinaris lectin. The chimeric lectin expressed in E.coli was found to bind mannosyl-bovine serum albumin and this binding was inhibitied by mannose. Then nonapeptide sequence was replaced with random amino acid sequences, which were introduced into cDNA coding BPA by use of polymerase chain reaction technique. The constructed lectin cDNA was inserted into lambda gtll or lambda foo expression vector, followed by expression in E.coli cells. In lambda gtll system, lectin was detected as fusion protein of beta-galactosidase. In lambda foo pharge, lectin molecule had not been detected yet.
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