Construction of an integration vector for use in the archaebacterium Methanococcus voltae and expression of a eubacterial resistance gene |
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Authors: | Petra Gernhardt Odile Possot Maryline Foglino Lionel Sibold and Albrecht Klein |
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Institution: | (1) Molekulargenetik, Fachbereich Biologie, Philipps-Universität, Postfach 1929, D-3550 Marburg, Germany;(2) Département des Biotechnologies, Institut Pasteur, Unité de Physiologie cellulaire et LIRA 1300 - CNRS, F-75724 Paris Cedex 15, France;(3) Present address: Laboratoire de Chimie Bactérienne, CNRS, PB 71, F-13277 Marseille Cedex 9, France |
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Abstract: | Summary An integration vector for use in Methanococcus voltae was constructed, based on the Escherichia coli vector pUC18. It carries the structural gene for puromycin transacetylase from Streptomyces alboniger, which is flanked by expression signals of M. voltae structural genes and hisA gene sequences of this bacterium. Transformed M. voltae cells are puromycin resistant. Several types of integration of the vector into the chromosome were found. Only one case was due to nonhomologous recombination. The integrated sequences were stable under selective pressure but were slowly lost in some cases in the absence of the selective drug. The vector could be excised from M. voltae chromosomal DNA, recircularized and transformed back into E. coli.The order of the other authors is not indicative of the relative importance of their experimental contributions which are considered to be equivalentWe mourn the loss of our colleague and friend Lionel Sibold, who died while this work was still in progress |
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Keywords: | Methanococcus voltae Transformation Integration vector Vector stability Expression of heterologous resistance gene |
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