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Transient gene expression in guard cell chloroplasts of tobacco using ArF excimer laser microablation

Paper ID Volume ID Publish Year Pages File Format Full-Text
21660 43233 2008 5 PDF Available
Title
Transient gene expression in guard cell chloroplasts of tobacco using ArF excimer laser microablation
Abstract

In this paper, we report a novel method for delivering genes into chloroplasts of tobacco cells using laser microablation. The plasmid pLD200-GFP was introduced into chloroplasts of Nicotiana tabacum cv. Xanthi guard cells and transient GFP expression was detected in the chloroplasts after 2–3 d of incubation. The technique uses an argon fluoride (ArF) excimer laser to perforate the cell surface in a 4 μm2 area in the presence of plasmid coated gold microparticles. Pretreatment of guard cells to promote stomatal closure prior to laser ablation resulted in a significant increase in the survival rate of cells and a transient expression rate of 2–3% in trial number basis was archived. Our method has unique advantages such as avoiding laborious pretreatments that adversely affect cell viability and specific delivery of transgenes into a desired cell in complex leaf tissue. This technique is a potential tool for cell specific transient gene expression studies for elucidation of gene regulation and expression.

Keywords
chloroplast transformation; guard cell; laser microablation
First Page Preview
Transient gene expression in guard cell chloroplasts of tobacco using ArF excimer laser microablation
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 106, Issue 2, August 2008, Pages 194–198
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering