Introduction of genes for leucine biosynthesis from Clostridium pasteurianum into C. acetobutylicum by cointegrate conjugal transfer |
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Authors: | John D Oultram Hazel Peck John K Brehm Daphne E Thompson Tracy J Swinfield and Nigel P Minton |
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Institution: | (1) Molecular Genetics Gruop, Division of Biotechnology, PHLS Centre for Applied Microbiology and Research, Porton Down, SP4 OJG Salisbury, Wiltshire, UK;(2) Present address: Department of Biological Sciences, Sheffield Polytechnic, Pond Street, S1 1WB Sheffield, United Kingdom |
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Abstract: | Summary A Clostridium pasteurianum gene bank was constructed in Escherichia coli, using plasmid pAT153, and several chromosomal fragments found which complemented both leuB and leuC mutations in auxotrophic E. coli K12 strains. No fragments capable of complementing leuA or leuD mutations were identified. Conjugal transfer of the LeuB/leuC genes from Bacillus subtilis into two different Leu-
C. acetobutylicum auxotrophic strains was elicited by their incorporation into a large plasmid cointegrate composed of the conjugal plasmid pAM 1 and a specially constructed gram-positive, replication-deficient plasmid, pMTL21 EC. Inheritance of the cointegrate plasmid restored one of the auxotrophic C. acetobutylicum strains to prototrophy. The second strain remained Leu-. |
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Keywords: | Leucine biosynthetic genes Cointegrate cloning vector Conjugal transfer Clostridium acetobutylicum pAM 1" target="_blank">gif" alt="beta" align="MIDDLE" BORDER="0">1 |
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