エレクトロポレーションの最適条件によるBucillus subtilisの形質転換とRuminococcus albusセルラーゼ遺伝子の発現

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タイトル別名
  • Electro-transformation of Bacillus subtilis for Expressing a Ruminococcus albus Cellulase Gene
  • エレクトロポレーション ノ サイテキ ジョウケン ニ ヨル Bacillus

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説明

Electroporation conditions for Bacillus subtilis transformation were optimized witha shuttle vector (pHY 300 PLK) for Escherichia coli and B. subtilis. The highest transformation efficiency (3.2×105 transformants per μg of plasmid DNA) of B. subtilis was obtained on platescontaining tetracycline, when the cells harvested at an early stationary phase were eleciroporated with double pulses of 300-400μsec in an electric field strength of 10-12kV/cm in the presence of plasmid DNA (0.05-0.06μg) and post-pulse-incubated at 37°C for 3h. A HindIII fragment containing an endoglucanase gene (egI) of Ruminococcus albus F-40 was put into the shuttle vector. With the resultant chimera plasmid, pHRA 101, B. subtilis was transformed using optimal electroporation conditions. This endoglucanase gene was expressed in B.subtilis.

収録刊行物

  • 日本畜産学会報

    日本畜産学会報 65 (5), 407-415, 1994

    公益社団法人 日本畜産学会

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