Hybrid genes in the analysis of transformation conditions: II. Transient expression vs stable transformation - analysis of parameters influencing gene expression levels and transformation efficiency

Negrutiu, Ioan; Dewulf, Jocelyne; Pietrzak, M.; Botterman, J.; Rietveld, E. M.; Wurzer-Figurelli, E. M.; Ye, De; Jacobs, M.

doi:10.1034/j.1399-3054.1990.790130.x

Cited by 9 publications

(17 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Transformation frequencies from 1% up to 12% have been reported by use of direct gene transfer in optimized systems where PEG has been included (Shillito et al 1985;Negrutiu et al 1987Negrutiu et al , 1990Paszkowski et al 1989). Compared with these values, the efficiency of pea transformation is still fairly low.…”

Section: Gus Analysis and Selection Of Transgenic Callusmentioning

confidence: 95%

“…Compared with these values, the efficiency of pea transformation is still fairly low. Transformation frequencies in moth bean (K6hler et al 1987a,b) and winged bean (Gill 1990), obtained by PEGmediated DNA uptake, are about the same as in electroporated pea, although transformation frequencies obtained by use of PEG have been shown to be higher than those obtained by electroporation in some plant species (Negrutiu et al 1990). As no protocols have so far been published on pea transformation by use of PEGmediated gene transfer, no comparisons are possible.…”

Section: Gus Analysis and Selection Of Transgenic Callusmentioning

confidence: 99%

“…The competence for transformation by use of direct gene transfer has been shown to be species-specific (Negrutiu et al 1987(Negrutiu et al , 1990. Moreover, the cultivar used affects the transformation frequencies in direct gene transfer experiments in e.g.…”

Section: Gus Analysis and Selection Of Transgenic Callusmentioning

confidence: 99%

“…Thus, a screening for cultivars with high competence for transformation would be in order in pea. However, as the results from transient expression studies rarely correlate with those from integrative transformation (Negrutiu et al 1990), the screening should be done on the level of stable transformation, instead of by studying transient expression only.…”

Section: Gus Analysis and Selection Of Transgenic Callusmentioning

confidence: 99%

See 3 more Smart Citations

Survival and growth of pea protoplasts after transformation by electroporation

Puonti‐Kaerlas

Ottosson

Eriksson

1992

Plant Cell Tiss Organ Cult

View full text Add to dashboard Cite

Protoplasts from two different pea cultivars, Belman and Filby, were stably transformed by direct gene transfer using electroporation. Transgenic calli could be obtained after selection, when hygromycin resistance was used as the selective trait introduced into the protoplasts, while no transformants were obtained when kanamycin resistance was used as selective marker in either of the two pea cultivars tested. The effect of the field strength on survival and division rates of the protoplasts was studied. Two different culture systems and osmotica were compared for induction of sustained divisions in and regeneration of transgenic callus from the protoplasts. The choice of the culture system had a considerable effect on the initial division frequency of the treated protoplasts, as well as on the later growth of the colonies. Transformation efficiency was monitored by histochemical GUS assay, and the transgenic nature of the calli selected for resistance against antibiotics was confirmed by DNA analysis.

show abstract

Section: Gus Analysis and Selection Of Transgenic Callusmentioning

confidence: 95%

Section: Gus Analysis and Selection Of Transgenic Callusmentioning

confidence: 99%

Section: Gus Analysis and Selection Of Transgenic Callusmentioning

confidence: 99%

Section: Gus Analysis and Selection Of Transgenic Callusmentioning

confidence: 99%

See 2 more Smart Citations

Survival and growth of pea protoplasts after transformation by electroporation

Puonti‐Kaerlas

Ottosson

Eriksson

1992

Plant Cell Tiss Organ Cult

View full text Add to dashboard Cite

show abstract

“…The efficiency of direct gene transfer (DGT) has been increased dramatically from 10-6 or lower to more than 10 -3 in tobacco [27]. In other species, such as the closely related Nicotianaplumbaginifolia, and despite the demonstration that the levels of transient gene expression are high and that transfection conditions were optimized according to the species [21,19], stable transformation frequencies remained one or two orders of magnitude lower. These results suggested that there were species-specific factors controlling the fate of the transforming DNA.…”

Section: Introductionmentioning

confidence: 99%

Hybrid genes in the analysis of transformation conditions. 3. Temporal/spatial fate of NPTII gene integration, its inheritance and factors affecting these processes in Nicotiana plumbaginifolia

et al. 1990

Self Cite

View full text Add to dashboard Cite

Freshly isolated haploid mesophyll protoplasts of Nicotiana plumbaginifolia were transformed for kanamycin resistance. In 38% of the 224 transformants analysed, transmission of the NPTII gene occurred as a homozygous trait, while 62% of the transformants were heterozygous for the trait. In the first case, the foreign DNA integration predominantly (95%) resulted in monogenic inheritance. The second group was characterized by a significant (46%) proportion of multiple insertions. However, there was no clear-cut difference in the integration pattern between the two groups. Furthermore, transformation rates were increased by 4- to 10-fold when transformed diploid protoplasts were treated with UV light or with 3-aminobenzamide. The number of insertion sites was also increased by these treatments. These results shed further light on the fate of the foreign DNA in transformed plants and on means to control or manipulate the integration event(s).

show abstract

Yeast RAS2 affects cell viability, mitotic division and transient gene expression in Nicotiana species

et al. 1990

View full text Add to dashboard Cite

Overexpression of the budding yeast RAS2 gene in Nicotiana plumbaginifolia cells revealed that RAS2 acted as 'suicide' gene in freshly isolated protoplasts from leaves and blocked cell proliferation in cell suspension-derived protoplasts. Among a series of genes tested (such as npt II, CDC35, PDE2), RAS2 was the only one to block the expression of the cat gene, as measured in a transient gene expression assay. Another ras gene, v-Ha-ras, had similar effects. Furthermore, the RAS2 effect was species-specific and depended on the modulation of hormonal metabolism in the transfected cells, while no differences were noticed between the normal and the activated val19 gene. Transfected plant cells are shown to synthesize a RAS2 protein of the same electrophoretic mobility as the yeast RAS2 product. The results are discussed in the broader context of the evolutionarily conserved ras genes involved in vital cellular functions.

show abstract

Hybrid genes in the analysis of transformation conditions: II. Transient expression vs stable transformation - analysis of parameters influencing gene expression levels and transformation efficiency

Cited by 9 publications

References 0 publications

Survival and growth of pea protoplasts after transformation by electroporation

Survival and growth of pea protoplasts after transformation by electroporation

Hybrid genes in the analysis of transformation conditions. 3. Temporal/spatial fate of NPTII gene integration, its inheritance and factors affecting these processes in Nicotiana plumbaginifolia

Yeast RAS2 affects cell viability, mitotic division and transient gene expression in Nicotiana species

Contact Info

Product

Resources

About