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Gene-Engineering Biotechnology
Cloning of the Zea maize genome DNA sequences capable of autonomous replication in Saccharomyces cerevisiae
1Shulzhenko V. N., 1Kordyum V. A.
  1. Institute of Molecular Biology and Genetics, Academy of Sciences of the Ukrainian SSR
    Kiev, USSR


DNA fragments of maize genome functioning as autonomously replicating sequences (ARS) in yeast were inserted into EcoRI-site of plasmid pYF40 bearing the His3 marker of yeast Saccharomyces cerevisiae. Two plasmids with inserts were selected which transformed strain His- S. cerevisiae LL20(leu2-3leu2-llhis3-llhis3-15) into His+ with a frequency of (1-3)·102 transformants per 1 u.g of plasmid DNA. ARS-plasmids are located in yeast cells in a nonintegrated form, that is confirmed by the isolation of plasmid DNA from them, with which both E. coli and S. cerevisiae cells are transformed. Size of cloned EcoRI DNA fragments is about 0.8 MD.


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