| Project/Area Number |
05555221
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
生物・生体工学
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| Research Institution | NARA INSTITUTE OF SCIENCE AND TECHNOLOGY (1994)
Osaka University (1993) |
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Principal Investigator |
SHINMYO Atsuhiko Nara Institute of Science and Technology, Graduate School of Bioscience, Professor, バイオサイエンス研究科, 教授 (30029235)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIDA Kazuya Osaka University, Department of Biotechnology, Assistant Professor, 工学部, 助手 (50252622)
SEKINE Masami Osaka University, Department of Biotechnology, Assistant Professor, 工学部, 助手 (70226653)
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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 |
¥5,300,000 (Direct Cost: ¥5,300,000)
Fiscal Year 1994: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥3,300,000 (Direct Cost: ¥3,300,000)
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| Keywords | tobacco cultured cells / horseradish peroxidase gene / wound-induction / cis-element / transcription factor / cucumber ascorbate oxidase gene / heat shock promoter / Arabidopsis thaliana / アラビドプシス / 熱ショックエレメント / 西洋ワサビペルオキシダーゼ / 傷害誘導シスエレメント / GUS発現 / プロトプラスト / CaMV35Sプロモーター |
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| Research Abstract |
We studied construction of a high expression system of foreign genes in tobacco cultured cells, Nicotiana tabacum L.cv.BY2, which can grow quite faster among plant cells (doubling time is 10-11h). A set of target genes introduced into tobacco cells must be placed under the control of strong promoters, enhancers, and cis-elements responding to operation conditions of plant cell culture. A promoter of horseradish peroxidase isozyme gene, prxC2, showed high activity in tobacco cells. Expression of the prxC2 gene was induced by wounding, and a cis-element, of which core sequence was CACGTG,for wound-induction was identified at-289 bp from the translation intiation point. A cDNA encoding a transcription factor associated with the cis-element was also cloned. The wound-induction system seems to be effective gene expression in the tobacco cultured cells. A promoter of ascorbate oxidase gene in cucumber was also showed high transcription activity.
A heat shock promoter of HSP18.2 gene of Arabidopsis thaliana was ligated to the structural gene for beta-glucuronidase (GUS), and introduced into chromosome of the tobacco cultured cells. Expression of the GUS gene was strongly induced when incubation temperature was shifted from 25ﾟC to37ﾟC.GUS mRNA was detected at 15 min after temperature shift and was maximum at 2 h. Maximun GUS activity was found at 4 h. This result indicated that the HSP18.2 promoter is a good candidate for expression of foreign genes by the control of incubation temperature of tobacco cultured cells.
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