Chromosome Research

, Volume 1, Issue 2, pp 107–115 | Cite as

Localization of seed protein genes on flow-sorted field bean chromosomes

  • Jiří Macas
  • Jaroslav Doležel
  • Sergio Lucretti
  • Uta Pich
  • Armin Meister
  • Jörg Fuchs
  • Ingo Schubert
Article

Abstract

Chromosomes from reconstructed field bean (Vicia faba L.) karyotypes were flow-sorted and the DNA was used for the physical localization of seed storage and nonstorage (USP) protein genes using PCR with sequence specific primers. The data were confirmed and refined by using DNA of microisolated chromosomes of other karyotypes as the target for PCR. The specificity of the PCR products was proved by restrictase digestion into fragments of predicted length or by reamplification using ‘nested’ primers. The genes are located within defined regions of chromosome I (USP=unknown seedprotein genes), II (vicilin genes, legumin B3 genes), III (legumin B4 genes), IV (pseudogenes ψ1) and V (legumin A genes and pseudogenes ψ1). Except for the pseudogene derived from the sequence of legumin B4 gene, all members of each gene family are located in one chromosome region exclusively. This approach proved to be useful for localizing genes that cannot be mapped genetically (due to the lack of allelic variants) and might be applied to integrate physical and genetic maps.

Key words

Vicia faba chromosome flow sorting gene mapping PCR seed protein genes 

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Copyright information

© Rapid Communications of Oxford Ltd 1993

Authors and Affiliations

  • Jiří Macas
    • 1
  • Jaroslav Doležel
    • 2
  • Sergio Lucretti
    • 3
  • Uta Pich
    • 1
  • Armin Meister
    • 1
  • Jörg Fuchs
    • 1
  • Ingo Schubert
    • 1
  1. 1.the Institute of Plant Genetics and Crop Plant ResearchGaterslebenGermany
  2. 2.the Institute of Experimental BotanyOlomoucCzech Republic
  3. 3.Agricultural R&D DepartmentENEA, CRE CasacciaRomeItaly

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