Genetic transformation of plants by protoplast electroporation.
This article describes an optimized protocol for the electroporation of tobacco mesophyll protoplasts together with notes and data on the effects of various parameters and suggestions for work with protoplasts of other species. In this protocol, electroporation is achieved by means of electrical pulses from a high-voltage, capacitive-discharge unit. Procedures are described for measurement of protoplast viability with Evan's blue, the detection of transient expression of CAT and GUS gene plasmid constructs, and for the recovery of stable transformants based on selection for kanamycin resistance.
Bates
G W
GW
Department of Biological Science, Florida State University, Tallahassee 32306.
D002470:Cell Survival; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics*; D004351:Drug Resistance / Q000235:genetics; D018274:Electroporation; D005070:Evans Blue; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics; D010944:Plants / Q000235:genetics*; D010947:Plants, Toxic; D010957:Plasmids; D011523:Protoplasts; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic
D018744:DNA, Plant; D011993:Recombinant Fusion Proteins; D005070:Evans Blue; D015500:Chloramphenicol O-Acetyltransferase; D005966:Glucuronidase