A vector for the site-directed,genomic integration of foreign DNA into soybean root-nodule bacteria |
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Authors: | Gonzalo Acuña Ariel Alvarez-Morales Matthias Hahn Hauke Hennecke |
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Institution: | (1) Mikrobiologisches Institut, Eidgenössische Technische Hochschule, ETH-Zentrum, Universitätstrasse 2, CH-8092 Zürich, Switzerland;(2) Present address: CINVESTAV-IPN, Unidad Irapuato, Apdo, Postal 629, 36500 Irapuato, Gto, Mexico |
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Abstract: | A non-essential DNA region carrying two different repeated sequences (RS 3 and RS 9) adjacent to a nitrogen fixation (nif) gene cluster has been identified previously in Bradyrhizobium japonicum strain 110. In closely related B. japonicum strains a similar genomic arrangement was found. We constructed a mobilizable plasmid vector carrying RS 3 and RS 9, and a kanamycin resistance cassette (nptII gene) plus suitable cloning sites inserted between the two repeated sequences. Using this vector (pRJ1035), stable integration of a lacZ gene fusion into the B. japonicum genomic RS region was achieved. The resulting strain yielded more than 10-fold higher -galactosidase activity in soybean root nodules as compared to a B. japonicum strain carrying the same lacZ fusion on a pRK290-based plasmid. |
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Keywords: | Bradyrhizobium gene transfer nitrogen fixation plasmid root-nodule symbiosis vector construction |
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