Transformation of Pseudomonas putida by Electroporation
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概要
- 論文の詳細を見る
The optimum electrotransformation conditions were determined for Pseudomonas putida PpY101 with plasmid pSUP104 (9.5kb) and pSR134 (18.6kb). Field strength was a very important parameter for electrotransformation efficiency. Optimum efficiencies (1.1x 10^5 transformants/μg DNA) with pSUP104 and pSR134 were obtained at a field strength of 12. 5 kV/cm, a time constant of about 4.5 ms (resistance setting of 200 Ω), a supercoiled DNA concentration of 100 ng/ml, and a cell concentration of 10^9/ml. Because the effciency obtained is high enough, electrotransformation is useful for the direct cloning of P. putida PpY101. No significant relationship between plasmid size and electrotransformation efficiency was observed. These efficiencies were about 4.5 times higher than those using the MgCl_2 method. Under these conditions, electrotransformation efficiencies of relaxed plasmid DNA treated with topoisomerase I and that linearized by EcoRI digestion were high.
- 社団法人日本農芸化学会の論文
- 1994-05-23
著者
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Uchiyama H
National Inst. Environmental Studies Ibaraki Jpn
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Uchiyama Hiroo
National Institute Of Environmental Studies
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Uchiyama Hiroo
Water And Soil Environment Division National Institute For Enviromental Studies
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Yagi Osami
Water and Soil Environment Division, National Institute for Environmental Studies
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Yagi Osami
Water And Soil Environmental Studies
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Ishizuka Kozo
Institute of Applied Biochemistry, University of Tsukuba
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Iwasaki K
Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat
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KURABAYASHI Teruyo
Institute of Applied Biochemistry, University of Tsukuba
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Ishizuka K
Univ. Tsukuba Ibaraki Jpn
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Ishizuka Kozo
Institute Of Applied Biochemistry The University Of Tsukuba
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Yagi O
Univ. Tokyo Tokyo Jpn
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Takamura Y
Department Of Bioresources Sciences School Of Agriculture Ibaraki University
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Iwasaki Kazuhiro
Regional and Community Environment Division, National Institute for Environmental Studies
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Takamura Yoshichika
Divisvion of Bioresource Chemistry, Faculty of Agriculture, Ibaraki University
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Iwasaki K
National Inst. For Environmental Studies Ibaraki Jpn
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Kikuchi T
Ibaraki Univ. Ibaraki‐ken Jpn
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