1993 Fiscal Year Final Research Report Summary
Development of cell-free transcription system for a study of tissue-specific transcriptional regulation in plant gene
| Project/Area Number |
03554027
|
|---|
| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
植物生理学
|
|---|
| Research Institution | Nagoya University |
|---|
Principal Investigator |
YAMAZAKI Ken-ichi Nagoya Univ., Sch.Agr.Sci., Assistant, 農学部, 助手 (40182480)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NATORI Yohei Nisshin Flour Milling Co., Ltd., Central Research Inst., Researcher, 中央研究所, 専門研究員
YAMAGUCHI Junji Nagoya Univ., Sch.Agr.Sci., Assistant, 農学部, 助手 (10183120)
|
|---|
| Project Period (FY) |
1991 – 1993
|
| Keywords | plant / gene expression / in vitro transcription / nuclear extract / RNA polymerase / transcription system |
|---|
| Research Abstract |
An assay system that allows the rapid, direct, and quantitative detection of promoter-dependent transcription in vitro with nuclear extracts from tobacco cells has now been developed. The transcriptional machinery from tobacco cell nuclei effectively and accurately transcribes specific genes, and the optimum conditions for such a system are described. The system consists of a protein raction extracted from the nuclei of suspension-cultured tobacco cells (TBY-2), substrates, and an exogenously added plasmid DNA template. Specific initiation at the promoter was determined by analysis of the sizes of transcripts on a polyacrylamide gel that contained 8 M urea. Our system using a plasmid DNA template, based essentially on a G-free sequence system, was developed to elucidate the function of plant trans-acting factors that influence the activity of RNA polymerase II.However, further modifications that allow us to elucidate tissue-specific transcriptional activation may be required from now on.
|
