Transfer of a minimal linear marker-free and vector-free smGFP cassette into soybean via ovary-drip transformation.
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.
Liu
Jianfeng
J
Department of Bioscience and Biotechnology, School of Environmental & Biological Science & Technology, Dalian University of Technology, Dalian, Liaoning, 116024, People's Republic China. ljf525128@yahoo.com.cn
Su
Qiao
Q
An
Lijia
L
Yang
Aifu
A
D035264:Flowers / Q000502:physiology*; D005819:Genetic Markers; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D016254:Mutagenesis, Insertional / Q000235:genetics; D030821:Plants, Genetically Modified / Q000502:physiology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D013025:Glycine max / Q000502:physiology*; D014170:Transformation, Genetic / Q000235:genetics*
D005819:Genetic Markers; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins