1992
DOI: 10.1007/bf02525170
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Effective vectors for transformation, expression of heterologous genes, and assaying transposon excision in transgenic plants

Abstract: Progress in plant molecular biology has depended heavily on the availability of effective vectors for plant cell transformation and heterologous expression. In this paper we describe the structures of a wide array of plasmids which have proved extremely effective in (a) plant transformation, (b) expression of heterologous genes and (c) assaying the activity of transposons in transgenic plants. Constructs that confer resistance to kanamycin, hygromycin, streptomycin, spectinomycin and phosphinotricin, or that c… Show more

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Cited by 287 publications
(227 citation statements)
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“…High molecular weight BAC DNA was purified by cesium chloride-ethidium bromide gradient centrifugation and was partially digested with Sau3AI, dephosphorylated, and size-fractionated on 0.8% low melting agarose gels. Fragments of approximately 15-20 kb were eluted from gels and inserted into the BamHI site of cosmid vector pSLJ755I5 (15). Ligated DNA was packaged in phage heads by using commercial extracts (Stratagene Gigapack III Gold) and used to transfect DH10B Escherichia coli cells (GIBCO).…”
Section: Methodsmentioning
confidence: 99%
“…High molecular weight BAC DNA was purified by cesium chloride-ethidium bromide gradient centrifugation and was partially digested with Sau3AI, dephosphorylated, and size-fractionated on 0.8% low melting agarose gels. Fragments of approximately 15-20 kb were eluted from gels and inserted into the BamHI site of cosmid vector pSLJ755I5 (15). Ligated DNA was packaged in phage heads by using commercial extracts (Stratagene Gigapack III Gold) and used to transfect DH10B Escherichia coli cells (GIBCO).…”
Section: Methodsmentioning
confidence: 99%
“…After digesting the DNA partially with Mbol, the ends of the fragments were incompletely filled in with dGTP and dATP. DNA fragments were then fractionated by centrifugation in a linear 1.25 to 5.0 M NaCl gradient, and DNA in the 20-kb range was ligated into the Xhol site (incompletely filled in with dTTP and dCTP) of the cosmid pDCL04541 (Jones et al, 1992). The ligation mix was packaged using Gigapack II (Stratagene, La Jolla, CA) and used to infect the Escherichia coli DH5a.…”
Section: Gene Cloningmentioning
confidence: 99%
“…The TAMU bacterial artificial chromosome (BAC) T31A11 was digested partially with HindIII and subcloned into the binary cosmid vector pSLJ755I5 (Jones et al, 1992). Partially digested products were size selected by electrophoresis in an 0.8% agarose gel, and fragments longer than 15 kb were recovered by using Gene Clean III (Bio 101, Vista, CA).…”
Section: Cosmid Subclones and Arabidopsis Transformationmentioning
confidence: 99%