Transformation of Soybean Embryogenic Cultures by Microprojectile Bombardment.

Cho, Myeong Je; Vodkin, Lila O.; Widholm, Jack M.

doi:10.5511/plantbiotechnology.14.11

Cited by 4 publications

(1 citation statement)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For PCR analysis, the gus gene primer set, 5′-AA-TTGATCAGCGTTGGTGG-3′ and 5′-GGTGTAGAGCATTACG-CTGC-3′, which yields a 0.45-kbp fragment inside of the gus gene (Anzai et al 1996), or the hpt gene primer set, 5′-CCTGAA-CTCACCGCGACG-3′ and 5′-AAGACCAATGCGGAGC ATAT-AC-3′, which generates a 0.81-kbp fragment inside of the hpt gene (Cho et al 1997), were used. PCR amplification reactions contained 20 ng of template DNA, 0.4 µM of each primer, 100 µM of a dNTP mixture, 1× Taq DNA polymerase reaction buffer and 1 U Taq DNA polymerase (Takara, Japan) in a 20 µl final volume.…”

Section: Polymerase Chain Reaction and Southern Blot Analysesmentioning

confidence: 99%

Agrobacterium-mediated production of transgenic plants of Muscari armeniacum Leichtl. ex Bak.

Suzuki

Nakano

2002

Plant Cell Rep

View full text Add to dashboard Cite

A system for the production of transgenic plants has been developed for the Liliaceous ornamental plant Muscari armeniacum Leichtl. ex Bak via Agrobacterium-mediated transformation of embryogenic cultures. Leaf-derived embryogenic cultures were co-cultivated with each of three A. tumefaciens strains, all of which harbored the binary vector carrying the neomycin phosphotransferase II (nptII), hygromycin phosphotransferase (hpt) and intron-containing β-glucuronidase (gus-intron) genes in the T-DNA region. Following co-cultivation, the embryogenic cultures were cultured on a medium containing 500 mg l -1 cefotaxime for 1 week followed by a medium containing 75 mg l -1 hygromycin in addition to cefotaxime. After 4-5 weeks, several hygromycin-resistant (Hyg r ) cell clusters were produced from the co-cultivated embryogenic cultures. The highest efficiency of production of Hyg r cell clusters was obtained when embryogenic cultures were inoculated with A. tumefaciens EHA101/pIG121Hm in the presence of 100 µM acetosyringone (AS) and 0.1% (v/v) of a surfactant (Tween20) followed by co-cultivation in the presence of 100 µM AS. Hyg r embryogenic cultures developed into complete plants via somatic embryogenesis, and most of them were verified to be transgenic by GUS histochemical assay and polymerase chain reaction analysis. Southern blot analysis revealed the integration of one to five copies of the transgene into the genome of transgenic plants, but most of them had one or two copies.

show abstract

Section: Polymerase Chain Reaction and Southern Blot Analysesmentioning

confidence: 99%