Bleomycin-induced gene expression in E.coli carrying blmA gene
| Project/Area Number |
05454570
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biological pharmacy
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| Research Institution | HIROSIMA UNIVERSITY |
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Principal Investigator |
SUGIYAMA Masanori School of Medicine, Professor, 医学部, 教授 (30106801)
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| Co-Investigator(Kenkyū-buntansha) |
INOUYE Yoshio School of Medicine, Associate Professor, 医学部, 助教授 (00136053)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥6,900,000 (Direct Cost: ¥6,900,000)
Fiscal Year 1994: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1993: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Keywords | Bleomycim / Inducer / beta-lactamase / Antitumor agent / インデューサー |
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| Research Abstract |
A bleomycin (Bm)-producing microorganism, Streptomyces verticillus ATCC15003 possesses two Bm resistance genes designated blmA and blmB.When one of these genes, designatedblmA,was inserted into pUC18 (or pUC19) plasmid and introduced into Escherichia coli TG1, the transformed cells expressed high level-resistance to Bm. In the present study we show that E.coli harboring the blmA-encoding pUC vector overproduced a 30-kDa protein when cultured in the presence of Bm. The Bm-induced protein was identified as the beta-lactamase encoded by the ampicillin resistance gene on the plasmid. In addition, we found that although the Bm family such as pepleomycin, phleomycin, cleomycin and liblomycin gave rise to beta-lactamase induction, bleomycininc acid which lacks terminal amine moiety in Bm molecule, did scarcely. We propose a hypothesis that Bm, an antibiotic inhibitor of DNA synthesis, acts as a an inducer (or an activator) for the expression of a specific gene in the presence of blmA.On the other hand, we constructed a chimeric plasmid designatd as intelligent vector pMAB70, which carried blmA and a tyrosinase structual gene put under the control of beta-lactamase promoter. E.coli harboring pMAB70 produced the melanine pigment, when cultured in the presence of Bm.
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Report
(3 results)
Research Products
(9 results)
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[Publications] Sugiyama, M., Thompson, C.J., Kumagai, T., Suzuki, K., Deblaere, R., Villarroel, N.and Davies, J.: "Characterisation by molecular cloning of two genes from Streptomyces verticillus encoding resitance to blemycin" Gene. 151. 11-16 (1994)
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