Summary
An autonomously replicating sequence (ars) of Nicotiana tabacum was cloned into the EcoRI site of YIp5, which consists of pBR322 and the yeast ura3 gene. Recombinant DNA was capable of transforming ura3- Saccaromyces cerevisiae YNN140 at 103–104 transformants per μg DNA. Transformants had a generation time of 3–4 h in the medium used for selection, in which approximately 30% of the cell retained the Ura+ phenotype after 24 h. The average copy number of hybrid plasmids was in the range of 10–20 molecules per cell. The size of the inserted DNAs from tobacco nuclear DNA was 4.9, 3.0 and 1.2 kilobase pairs (kbp), of which the 1.2 kbp insert hybridized to several bands of EcoRI-digested nuclear DNA. An EcoRI-generated 1.7 kbp ars fragment was cloned from tobacco chloroplast DNA.
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Abbreviations
- kbp:
-
kilobase pairs
- rDNA:
-
DNA coding for RNA
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Communicated by H. Saedler
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Uchimiya, H., Ohtani, T., Ohgawara, T. et al. Molecular cloning of tobacco chromosomal and chloroplast DNA segments capable of replication in yeast. Molec Gen Genet 192, 1–4 (1983). https://doi.org/10.1007/BF00327638
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DOI: https://doi.org/10.1007/BF00327638