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1995 | 36 | 1 | 1-10
Article title

Transient expression assay for the optimization of direct gene transfer into cucumber meristem protoplasts by electroporation

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EN
Abstracts
EN
The paper obtained a new way of viable and very homogenous cucumber protoplasts.Protoplasts from cells formed in the shoot tip meristem culture were isolated from suspension.Plasmid pBI121 was introduced using imulse electric field.Effectiveness of transformation process was determinated by transient expression of ?-glucoronidase (GUS) gene, controlled by promotor 35S.The ativity of ?-glucoronidase enzyme as a product of GUS reporter gene was estimated by fluorometric method (Jefferson 1987).Parameters of electroporatian process were optimized.The transient expression of GUS gene was measured 24 h after electroporation.The highest effectiveness of transformation process was achieved using three electric impulses at the initial voltage of 250-350V at 10-sec.intervals as a results of discharging a 140 ?F capacitor and 50-70 ?g x cm/1000 plasmid DNA in the presence of 50 ?g x cm/1000 carrier DNA.The system presented is an effective method of exogenic DNA transfer, which is indicated by a high transient expression of the reporter gene.In comparison to Agrobacterium tumefaciens and A.rhizogenes, this alternative method of gene transfer can be used for obtaining transgenic cucumber plants.
Discipline
Year
Volume
36
Issue
1
Pages
1-10
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author
author
author
author
References
Document Type
article
Publication order reference
W.Burza
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YADDA identifier
bwmeta1.element.element-from-psjc-1de69966-f644-3a78-a00d-7ee4db037f17
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