Stabilization of a Recombinat Plasmid in Yeast
スポンサーリンク
概要
- 論文の詳細を見る
When Saccharomyces cerevisiae AH22R^--2075 harboring pGLD p31-RcT which encodes a modified hepatitis B virus-surface antigen P31 (rHBsAg P31 mutein) coding gene and β-isopropylmalate dehydrogenase gene (LEU2) was cultivated on a large scale in a leucine-lacking medium, rHBsAg non-producing colonies appeared at high frequency. Restriction enzyme analysis and Southern blot analysis with LEU2 probe revealed that the cells from these non-producting colonies had lost the expression plasmid, pGLD p31-RcT, and had unusual 2 μm DNA containing the LEU2 gene. This was caused by homologous recombination between 2 μm DNA and pGLD p31-RcT which contains a 2 μm DNA fragment and the LEU2 gene. Stabilization of the recombinant plasmid was achieved by the following three approaches : (i) reconstruction of the expression plasmid by separeting the LEU 2 gene from the 2 μm DNA fragment on the plasmid; (ii) selection of plasmid-stabilized clones by taking advantage of the differences in growth rate between producing and non-producing clones; and (iii) addition of a high concentration of L-histidine to the medium.
- 社団法人日本生物工学会の論文
- 1992-09-25
著者
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Kuriyama Shinichi
Division Of Epidemiology Department Of Public Health And Forensic Medicine Tohoku University Graduat
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KURIYAMA MASATO
Microbiology Research Laboratories
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KITANO KAZUAKI
Microbiology Research Laboratories
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MORITA SHIGERU
Microbiology Research Laboratories, Research and Development Division, Takeda Chemical Industries, L
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ASAKAWA NAOKI
(Present address) Hikari Plant, Takeda Chemical industries, Ltd.,
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NAKATSU MASANORI
Microbiology Research Laboratories, Research and Development Division, Takeda Chemical Industires, L
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Kitano Kazuaki
Microbiological Research Laboratories Central Research Division Takeda Chemical Industries Ltd.
関連論文
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- Stabilization of a Recombinat Plasmid in Yeast
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