1998 Fiscal Year Final Research Report Summary
Large-scale isolation of plant genes for the use in molecular breeding
| Project/Area Number |
08556002
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (A)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 展開研究 |
|---|
| Research Field |
Breeding science
|
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
UCHIMIYA Hirofumi The University of Tokyo, Institute of Molecular and Cellular Biosciences, Professor, 分子細胞生物学研究所, 教授 (50142229)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
TSUKAYA Hirokazu The University of Tokyo, Institute of Molecular and Cellular Biosciences, Profes, 分子細胞生物学研究所, 助手 (90260512)
UMEDA Masaaki The University of Tokyo, Institute of Molecular and Cellular Biosciences, Assist, 分子細胞生物学研究所, 助手 (80221810)
|
|---|
| Project Period (FY) |
1996 – 1998
|
| Keywords | Rice / Arabidopsis / EST / Breeding / Transgenic plants / Homeostasis / Metabolic pathway / Database |
|---|
| Research Abstract |
This project aimed to develop a system to characterize plant genes in a large-scale. Genes involved in metabolic pathways were intensively characterized for the use in molecular breeding in agriculture. To identify a specific gene, one can purify the target protein by repeated chromatography steps and make a peptide corresponding to a particular region. Another way is to search for a specific domain which is conserved in related proteins among several organisms and synthesize degenerate primers. However, to characterize numerous genes, such methods cannot be applied because of the loss of information of expressed genes.
In this project, cDNAs of rice and Arabidopsis were randomly isolated and characterized by searching for similarity to other proteins. mRNA was isolated from suspension culture or different tissues, and cDNAs were directly cloned into the pUC vector. cDNA fragments were randomly isolated and subjected to sequence analysis using the GenBank database. To efficiently identify a related gene or protein, we also developed a software system for identification of amino acid similarities. Characterized cDNAs were categorized by predicted functions and further subjected to search for conserved domains.
Based on the information of typical domains found in categorized protein family, degenerate primers were synthesized, and other related genes including such domains were fished out by conventional screening of rice and Arabidopsis cDNA libraries. Characterization of these genes provided us a series of candidates which may be useful for making transgenic plants, that are stress-tolerant, disease-resistant, or producing a high yield. Therefore, these genes are certainly powerful tools for molecular breeding in the field of agriculture.
|
