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 Previous Article

Applied and Environmental Microbiology, June 1999, p. 2802-2804, Vol. 65, No. 6
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

High-Efficiency Transformation of Rhizobium leguminosarum by Electroporation

Bindu Garg, Romesh C. Dogra,* and Parveen K. Sharma

Department of Microbiology, College of Basic Sciences and Humanities, CCS Haryana Agricultural University, Hisar 125 004, India

Received 4 December 1998/Accepted 23 March 1999

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, 108 transformants/µg 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 × 108 transformants/µg of DNA) was obtained with 0.5 µg of DNA/ml of cell suspension, and a further increase in the DNA concentration resulted in a decline in transformation efficiency.

* Corresponding author. Mailing address: Department of Microbiology, College of Basic Sciences & Humanities, CCS Haryana Agricultural University, Hisar 125 004, India. Phone: 91-1662-37721, ext. 4292 (O). Fax: 91-1662-34952. E-mail: hau{at}hau.ren.nic.in.

Applied and Environmental Microbiology, June 1999, p. 2802-2804, Vol. 65, No. 6
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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