1989
DOI: 10.1104/pp.91.1.440
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Genetic Transformation of Maize Cells by Particle Bombardment

Abstract: Intact maize cells were bombarded with microprojectiles bearing plasmid DNA coding for selectable (neomycin phosphotransferase [NPT II]) and screenable (,l-glucuronidase [GUS]) marker genes. Kanamycin-resistant calli were selected from bombarded cells, and these calli carried copies of the NPT II and GUS genes as determined by Southern blot analysis. All MATERIALS AND METHODS PlasmidsThe plasmid pNGI (8.2 kb) contains both a neomycin phosphotransferase II (NPT II) and a 3-glucuronidase (GUS) gene cloned in … Show more

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Cited by 142 publications
(54 citation statements)
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“…Particle bombardments of DNA constructs and subsequent tissue culture steps were conducted essentially as described (Klein et al, 1989;Bowen, 1992). PCR and DNA gel blot analyses were conducted on resistant calli and subsequently on selected transformed plants to confirm transgene incorporation.…”
Section: Transformation Methodsmentioning
confidence: 99%
“…Particle bombardments of DNA constructs and subsequent tissue culture steps were conducted essentially as described (Klein et al, 1989;Bowen, 1992). PCR and DNA gel blot analyses were conducted on resistant calli and subsequently on selected transformed plants to confirm transgene incorporation.…”
Section: Transformation Methodsmentioning
confidence: 99%
“…The first successful method was microparticle bombardment (Klein et al, 1988a(Klein et al, , 1988b(Klein et al, , 1989. The bombardment method has been adopted widely by many researchers for use on different maize tissue types and on other crops (Fromm et al, 1990;Gordon-Kamm et al, 1990;Armstrong, 1999).…”
mentioning
confidence: 99%
“…Previous efforts to introduce pBI121.1 DNA biolistically into unfractionated yellow-poplar suspension cultures resulted in transient expression of GUS, but not the recovery of kanamycin-resistant stable transformants (31). Sieving the cultures through 140-,um mesh enriched for small clusters of meristematic cells A (7,8,13,20 Transgenic yellow-poplar somatic embryos exhibited GUS expression in histochemical assays, indicating that substrate penetration was not a problem in differentiated tissues. As the somatic embryos matured into plantlets, there was preferential expression of GUS in leaves over roots.…”
Section: Discussionmentioning
confidence: 96%
“…A fluorometric assay of GUS activity was performed using the substrate 4-methyl-umbelliferyl-3-D-glucuronide (1 1 Histochemical detection of GUS expression with the substrate X-gluc generally followed the protocol of Klein et al (13). Color development in transformed calli and somatic embryos was usually complete within 2 h. Heterogeneously staining callus cells were incubated overnight at 25°C in the presence of X-gluc, and blue and white cell clusters were separated under a dissecting scope.…”
Section: Gus Assaysmentioning
confidence: 99%