High-efficiency transformation of Rhizobium leguminosarum by electroporation. |
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| Authors: | B Garg R C Dogra P K Sharma |
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| Institution: | Department of Microbiology, College of Basic Sciences and Humanities, CCS Haryana Agricultural University, Hisar 125 004, India. |
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| Abstract: | Electrotransformation of Rhizobium leguminosarum was successfully carried out with a 15.1-kb plasmid, pMP154 (Cmr), containing a nodABC-lacZ fusion by electroporation. The maximum transformation efficiency, 10(8) transformants/microg of DNA, was achieved at a field strength of 14 kV/cm with a pulse of 7.3 ms (186 Omega). The number of transformants was found to increase with increasing cell density, with no sign of saturation. In relation to DNA dosage, the maximum transformation efficiency (5.8 x 10(8) transformants/microg of DNA) was obtained with 0.5 microg of DNA/ml of cell suspension, and a further increase in the DNA concentration resulted in a decline in transformation efficiency. |
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