Transfer of a minimal linear marker-free and vector-free smGFP cassette into soybean via ovary-drip transformation.

Abstract

A tissue culture-independent plant transformation method, called ovary-drip transformation, was established in which a minimal linear gene cassette [35S CaMV promoter, open reading frame of soluble modified green fluorescent protein (smGFP), and NOS terminator] was transformed into soybean. The method is characterized by directly dripping a DNA solution, which is supplemented with a surfactant, onto the ovary wound 6-8 h after self-pollination. The growth of the pollen tube was measured after self-pollination. The movement of smGFP across the passageway toward the embryo sac was monitored using fluorescein isothiocyanate-labeled DNA. The transformation frequency reached 3.2% by PCR analysis. Southern analysis of the primary transformants denoted the integration of a single site smGFP. The transgenic plants exhibited a high level of smGFP expression which was visible in the immature embryos of the transgenic soybean.

• Authors: Jianfeng Liu, Qiao Su, Lijia An, Aifu Yang
• Journal: Biotechnology letters (Biotechnol Lett) Vol. 31 Issue 2 Pg. 295-303 (Feb 2009) ISSN: 1573-6776 [Electronic] Netherlands
• PMID: 18815733 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Genetic Markers
• Recombinant Proteins
• Green Fluorescent Proteins

Topics

• Flowers (physiology)
• Genetic Markers
• Genetic Vectors
• Green Fluorescent Proteins (genetics, metabolism)
• Mutagenesis, Insertional (genetics)
• Plants, Genetically Modified (physiology)
• Recombinant Proteins (genetics, metabolism)
• Glycine max (physiology)
• Transformation, Genetic (genetics)