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Culture of Embryonic Mouse Cochlear Explants and Gene Transfer by Electroporation
Published on: January 12, 2015
Gene transfer into isolated and cultured tobacco zygotes by a specially designed device for electroporation
1Research Center for Developmental Biology, College of Life Science, Wuhan University, Wuhan 430072, China e-mail: hyyang@whu.edu.cn Fax: +86-27-87646010, , , , , , CN.
Abstract:
We have established a technique for isolating, culturing and transforming tobacco zygotes. Zygotes were isolated by microdissection or enzymatic maceration from fertilized embryo sacs. Viable zygotes cocultured with mesophyll protoplasts underwent first division after 3 days of culture. Zygotes isolated by microdissection underwent a higher frequency of first division (61.2%) than those isolated by enzymatic maceration (30.5%). Globular embryos were formed only from microdissected zygotes, at a frequency of 8.7% after 1-2 weeks in culture. An efficient millicell device for the electroporation of DNA into zygotes was established. The electroporated zygotes divided in vitro at a frequency of 54.6% and developed into proembryos. Introduced GFP gene constructs showed transient expression in about 2.6% of the electroporated tobacco zygotes.
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