Transient expression of electroporated gene in leaf protoplasts of indica rice and influence of template topology and vector sequences

Chaudhury, A. ; Maheshwari, S. C. ; Tyagi, A. K. (1993) Transient expression of electroporated gene in leaf protoplasts of indica rice and influence of template topology and vector sequences Physiologia Plantarum, 89 (4). pp. 842-846. ISSN 0031-9317

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Official URL: http://www3.interscience.wiley.com/journal/1192959...

Related URL: http://dx.doi.org/10.1111/j.1399-3054.1993.tb05294.x

Abstract

Electroporation was used for gene delivery and evaluation of various parameters affecting transient expression of a gene for β-glucuronidase (gus) in leaf protoplasts of Oryza sativa var. Basmati-370. Transient expression was found to be dependent on voltage, capacitance, amount of plasmid and carrier DNA as well as period of culture. Maximum GUS activity was obtained when a 150 ms pulse at 300 V cm-1 and 200 μF was applied to the protoplasts (l-2×106ml-1) in an electroporation buffer containing 20 μg of plasmid and 30 μg of calf thymus DNA, assayed 48 h after electroporation. DNA topology did not influence expression of the gene as both linear and supercoiled templates resulted in similar activities, but a 4-fold decrease in expression was observed if the gene was excised, reflecting the positive influence of vector sequences on gene expression.

Item Type:Article
Source:Copyright of this article belongs to Scandinavian Society for Plant Physiology.
Keywords:Electroporation; GUS Assay; Indica Rice; Oryza sativa; Protoplasts; Transient Gene Expression; Template Topology; Vector Sequences
ID Code:18500
Deposited On:17 Nov 2010 09:11
Last Modified:06 Jun 2011 11:33

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