• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to project page

2001 Fiscal Year Final Research Report Summary

Direct cloning of genes for crop cultivation approached by positional Information

Research Project

Project/Area Number 11460005
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Breeding science
Research InstitutionKihara Institute for Biological Research, Yokohama City University

Principal Investigator

OGIHATA Yasunari  Kihara Institute for Biological Research, Yokohama City University, Associate Professor, 木原生物学研究所, 助教授 (40185533)

Project Period (FY) 1999 – 2001
KeywordsPolyploid wheat / Large size DNA / Genomic library / Gene cloning / Q gene / Differential Display / Chromosome deletion lines / Mapping
Research Abstract

The amplified restriction fragment length polymorphism (AFLP)-based mRNA fingerprinting (AMF) method was applied to the cloning of the Q gene of common wheat. Using the AMF method, we comapred the fingerprints of mRNA samples extracted from the young spikes of Triticum aestivum cv. Chines Spring (CS) carrying the Q gene to those of a chloromsome deletion line of CS. Approximately 12200 fragments were produced after PCR with 256 primer combinations. Of these, 92 fragmentsw were differentially expressed between CS and deletion line. Northern nad Southern analyses showed that 16 fragments gave specific or relatively stronger transcript signals in CS, and these clones were present in single copy or in low copy numbers in the wheat genomes. Four clones were genetically mapped to the region deleted. Subsequently, one clone was mapped at the same locus as the Q gene, indicating that the clone corresponds to the Q gene or is tightly linked to it. However, further mapping experiment suggested that the clone was segregated from the Q gene.
Large-insert genomic DNA libraries of common wheat were constructed in a newly developsed transformation-competent artificial chromosome (TAC) vector, which accepts and maintains large genomic DNA fragments stably in both Escerichia coli and Agrobacterium tumefaciens. The vector contains the cis sequence required for Agrobacterium-mediated gene transfer into grasses. TAC clones containing genes(s) of interest were immediately transferred into grass genomes with the Agjobacterium system. TAC libraries constructed canbe used to isolate genomic clones containing target genes, and to carry out genome walking for positional cloning.

  • Research Products

    (5 results)

All Other

All Publications (5 results)

  • [Publications] Kojima, T., Y.Halra, S.lida, Y.Ogihara: "Direct isolation of differentially expressed genes from a specific chromosome region of common wheat : application of the AMF methodin combination with a deletion line of wheat"Moleculer and General Genetics. 263. 635-641 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Liu, YG, K.Nagaki, M.Fujita, K.Kawaura, M.Uozumi, T.Ogihara: "Development of an efficient maintenance and screening system for large-insert genomic libraries of hexaploid wheat in a TAC vector"Plant Journal. 23. 687-695 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 荻原保成: "コムギ(ムギ類)のゲノムとDNAレポジトリー"中辻憲夫(共立出版株式会社). 5 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T. Kojima, Y. Habu, S. Iida, Y. Ogihara: "Direct isolation of differentially expressed genes from a specific chromosome region of common wheat: Application of the amplified fragemtn length polymorphism-based mRNA fingerprinting (AMF) method in combination with a deletion line of wheat"Molecular and General Genetics. 263. 635-641 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.-G. Liu, K. Nagaki, M. Fujita, K. Kawamura, M. Uozumi, Y. Ogihara: "Development of an efficient maintenance and screening system for large-insert genomic DNA libraries of hexaploid wheat in a transformation-competent artificial chrommosome (TAC) vectors"Plant Journal. 23. 687-695 (2001)

    • Description
      「研究成果報告書概要(欧文)」より

URL: 

Published: 2003-09-17  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi