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Molecular Biology and Gene Regulation

Transient Gene Expression in Protoplasts of Phaseolus vulgaris Isolated from a Cell Suspension Culture ¹

Unidad de Biología Molecular y Biotecnología Vegetal, Universidad Nacional Autónoma de México Apdo Postal 565-A, Cuernavaca Morelos México, Department of Biological Sciences, Stanford University, Stanford, California 94305-5020

A cell suspension culture of Phaseolus vulgaris cv Negro jamapa was established. Protoplasts isolated from this culture are suitable for transient gene expression studies after DNA transfer by either electroporation or treatment with polyethylene glycol. The optimized conditions for both methods were established, taking into consideration expression and cell viability. We have investigated two features of posttranscriptional gene regulation. We demonstrate that inclusion of the RNA leader sequence, omega (), derived from tobacco mosaic virus, and a dicot intron in the leader region together enhance gene expression in this legume species about 10-fold.

¹ This work was supported support by the Rockefeller Foundation and a DGAPA grant from the National University of México.

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