Direct DNA transfer to plant cells

Davey, M. R.; Rech, E. L.; Mulligan, B. J.

doi:10.1007/bf00025315

Cited by 89 publications

(26 citation statements)

References 96 publications

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“…integration (Paszkowski et al, 1984;Davey et al, 1989;Saul and Potrykus, 1990;Christou, 1992). When the plant SAR plasmid was electroporated into tobacco NT-1 protoplasts before G U S assay 20 hr later, we observed an approximately threefold increase in GUS gene expression.…”

Section: Expression Levels In Stable Transformantsmentioning

confidence: 58%

High-level transgene expression in plant cells: effects of a strong scaffold attachment region from tobacco.

et al. 1996

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We have previously shown that yeast scaffold attachment regions (SARs) flanking a chimeric 0-glucuronidase (GUS) reporter gene increased per-copy expression levels by 24-fold in tobacco suspension cell lines stably transformed by microprojectile bombardment. In this study, we examined the effect of a DNA fragment originally identified in a tobacco genomic clone by its activity in an in vitro binding assay. The tobacco SAR has much greater scaffold binding affinity than does the yeast SAR, and tobacco cell lines stably transformed with constructs containing the tobacco SAR accumulated greater than fivefold more GUS enzyme activity than did lines transformed with the yeast SAR construct. Relative to the control construct, flanking the GUS gene with plant SARs increased overall expression per transgene copy by almost 140-fold.In transient expression assays, the same construct increased expression only approximately threefold relative to a control without SARs, indicating that the full SAR effect requires integration into chromosomal DNA. GUS activity in individual stable transformants was not simply proportional to transgene copy number, and the SAR effect was maximal in cell lines with fewer than 4 0 transgene copies per tobacco genome. Lines with significantly higher copy numbers showed greatly reduced expression relative to the low-copy-number lines. Our results indicate that strong SARs flanking a transgene greatly increase expression without eliminating variation between transformants. We propose that SARs dramatically reduce the severity or likelihood of homology-dependent gene silencing in cells with small numbers of transgenes but do not prevent silencing of transgenes present in many copies.

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Section: Expression Levels In Stable Transformantsmentioning

confidence: 58%

High-level transgene expression in plant cells: effects of a strong scaffold attachment region from tobacco.

et al. 1996

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“…Recent progress in plant cell biology and molecular biology allows the use of newly developed techniques to complement the work of conventional plant breeding [9,24]. The Ti plasmids of Agrobacterium tumefaciens have been converted into useful vectors for dicotyledonous plant transformation [2].…”

Section: Introductionmentioning

confidence: 99%

Fertile, transgenicTriticale ( �Triticosecale Wittmack)

et al. 1995

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Fertile transgenic Triticale ( x Triticosecale Wittmack) plants expressing the/?-glucuronidase (uidA) and phosphinothricin acetyltransferase (bar) genes were obtained after microprojectile bombardment of scutellar tissue with the plasmid pDB1 containing the uidA gene under the control of the actin-1 promoter (Act 1) from rice and the selectable marker gene bar under the control of the CaMV 35 S promoter. From 465 bombarded scutella about 4000 plantlets were regenerated; 300 plants survived the selection. These regenerants were screened for enzyme activity by the histological GUS assay and by spraying the plants with a herbicide (Basta). Twenty-five regenerants showed GUS activity and survived repeated Basta spraying. Southern blot analysis showed the presence of both marker genes introduced into the genome of analysed plants.All transgenic plants were fertile. They were grown to maturity and set seed. Pollen and progeny analyses provided evidence for inheritance of the introduced genes to the next generation.

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“…Direct DNA transfer techniques range from transfection of protoplast by electroporation or chemical methods to laser micropuncture of isolated cells [26,27]. The most common method is particle bombardment, also known as the biolistic method, a combination of biological and ballistic.…”

Section: Stable Nuclear Transformationmentioning

confidence: 99%

Virus-like particles production in green plants

Santi¹,

Huang²,

Mason³

2006

Methods

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Viruses like particles (VLPs), assembled from capsid structural subunits of several different viruses, have found a number of biomedical applications such as vaccines and novel delivery systems for nucleic acids and small molecules. Production of recombinant proteins in different plant systems has been intensely investigated and improved upon in the last two decades. Plant derived antibodies, vaccines, and microbicides have received great attention and shown immense promise. In the case of mucosal vaccines, orally delivered plant produced VLPs require minimal processing of the plant tissue, thus offering an inexpensive and safe alternative to more conventional live attenuated and killed virus vaccines. For other applications which require higher level of purification, recent progress in expression levels using plant viral vectors have shown that plants can compete with traditional fermentation systems. In this review the different methods used in the production of VLPs in green plants are described. Specific examples of expression, assembly, and immunogenicity of several plant-derived VLPs are presented.

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Direct DNA transfer to plant cells

Cited by 89 publications

References 96 publications

High-level transgene expression in plant cells: effects of a strong scaffold attachment region from tobacco.

High-level transgene expression in plant cells: effects of a strong scaffold attachment region from tobacco.

Fertile, transgenicTriticale ( �Triticosecale Wittmack)

Virus-like particles production in green plants

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