ISSN:
1432-203X
Keywords:
Cucumis
;
Cucumber
;
Protoplast
;
Transient expression
;
Tomato ringspot virus
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary A protocol was developed for the preparation of Cucumis sativus var Straight 8 protoplasts that incorporates a two-step Ficoll® gradient and results in a high percentage of viable, debris-free protoplasts suitable for the transient expression of foreign genes. Polyethylene glycol and electroporation were compared for their effect on protoplast transfection with commonly used reporter genes. Using a polyethylene glycol method, cucumber protoplasts transfected with a plasmid containing the β-glucuronidase gene showed high expression levels, while protoplasts transfected with a plasmid containing the chloramphenicol acetyl transferase gene showed levels of activity that were barely distinguishable from mock-transfected controls. Tomato ringspot virus genomic RNA was also transfected into the protoplasts, and the assembly of viral particles was confirmed.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00231947
Permalink