T-DNA Binary Vectors and Systems

Lee, Lan‐Ying; Gelvin, Stanton B.

doi:10.1104/pp.107.113001

Cited by 198 publications

(120 citation statements)

References 103 publications

Supporting

Mentioning

113

Contrasting

Unclassified

Order By: Relevance

“…The pSAT vectors (Lee and Gelvin, 2008) used for BiFC were kindly provided by Stanton Gelvin (Purdue University). The open reading frames of snapdragon GPPS.SSU and tobacco GGPPS-like proteins were fused in frame and upstream of the N-terminal half of EYFP in pSAT4-nEYFP-N1 and the C-terminal half of EYFP in pSAT6-cEYFP-C1, respectively, in the NcoI-BamHI cloning sites of both vectors.…”

Section: Bifcmentioning

confidence: 99%

The Small Subunit of Snapdragon Geranyl Diphosphate Synthase Modifies the Chain Length Specificity of Tobacco Geranylgeranyl Diphosphate Synthase in Planta

et al. 2009

View full text Add to dashboard Cite

Geranyl diphosphate (GPP), the precursor of many monoterpene end products, is synthesized in plastids by a condensation of dimethylallyl diphosphate and isopentenyl diphosphate (IPP) in a reaction catalyzed by homodimeric or heterodimeric GPP synthase (GPPS). In the heterodimeric enzymes, a noncatalytic small subunit (GPPS.SSU) determines the product specificity of the catalytic large subunit, which may be either an active geranylgeranyl diphosphate synthase (GGPPS) or an inactive GGPPS-like protein. Here, we show that expression of snapdragon (Antirrhinum majus) GPPS.SSU in tobacco (Nicotiana tabacum) plants increased the total GPPS activity and monoterpene emission from leaves and flowers, indicating that the introduced catalytically inactive GPPS.SSU found endogenous large subunit partner(s) and formed an active snapdragon/tobacco GPPS in planta. Bimolecular fluorescence complementation and in vitro enzyme analysis of individual and hybrid proteins revealed that two of four GGPPS-like candidates from tobacco EST databases encode bona fide GGPPS that can interact with snapdragon GPPS.SSU and form a functional GPPS enzyme in plastids. The formation of chimeric GPPS in transgenic plants also resulted in leaf chlorosis, increased light sensitivity, and dwarfism due to decreased levels of chlorophylls, carotenoids, and gibberellins. In addition, these transgenic plants had reduced levels of sesquiterpene emission, suggesting that the export of isoprenoid intermediates from the plastids into the cytosol was decreased. These results provide genetic evidence that GPPS.SSU modifies the chain length specificity of phylogenetically distant GGPPS and can modulate IPP flux distribution between GPP and GGPP synthesis in planta.

show abstract

Section: Bifcmentioning

confidence: 99%

The Small Subunit of Snapdragon Geranyl Diphosphate Synthase Modifies the Chain Length Specificity of Tobacco Geranylgeranyl Diphosphate Synthase in Planta

et al. 2009

View full text Add to dashboard Cite

show abstract

“…However, the pPZP-based destination vectors are not suited for transformation of A. tumefaciens since the most commonly used strains LBA4404, EHA101 and 105 of A. tumefaciens contain the avirulent Ti plasmid with the streptomycin-resistance marker. In addition, some strains of A. tumefaciens are reported to be resistant to low levels of spectinomycin [10]. Thus, the introduction of pPZP vectors to A. tumefaciens is not assured using simple antibiotic selection for streptomycin or spectinomycin, and needs additional steps to circumvent the difficulty.…”

Section: Introductionmentioning

confidence: 99%

“…This provides a practical advantage for using the tetracycline-based pLSU binary vectors as destination vectors for efficient cloning of multiple fragments to create expression vectors. The C58 strain of A. tumefaciens has a tetracycline-resistance determinant and is resistant to low levels of tetracycline [10].…”

Section: Introductionmentioning

confidence: 99%

Tetracycline-Based Binary Ti Vectors pLSU with Efficient Cloning by the Gateway Technology for <i>Agrobacterium tumefaciens</i>-Mediated Transformation of Higher Plants

Lee¹,

Su²,

Lasserre³

et al. 2013

AJPS

View full text Add to dashboard Cite

We constructed small high-yielding binary Ti vectors with a bacterial tetracycline resistance gene to facilitate efficient cloning afforded by the Gateway Technology (Invitrogen) for Agrobacterium tumefaciens-mediated transformation of higher plants. The Gateway Technology vectors are kanamycin-based, thus tetracycline-based destination and expression vectors are easily selected for the antibiotic resistance in the Escherichia coli media. We reduced the size of the tetracycline resistance gene TetC from pBR322 to 1468 bp containing 1191 bp of the coding region, 93 bp of 5'-upstream, and 184 bp 3'-downstream region. The final size of binary Ti vector skeleton pLSU11 is 5034 bp. pLSU12 and 13 have the kanamycin resistance NPTII gene as a plant-selectable marker. pLSU14 and 15 contain the hygromycin resistance HPH gene as a selection marker. pLSU13 and 15 also have the β-glucuronidase (GUS) reporter gene in addition to the plant selection marker. We also constructed a mobilizable version of tetracycline-based binary Ti vector pLSU16 in which the mob function of ColE1 replicon was maintained for mobilization of the binary vector from E. coli to A. tumefaciens by tri-parental mating. The final size of binary Ti vector skeleton pLSU16 is 5580 bp. New tetracycline-based binary Ti vectors pLSU12 were found as effective as kanamycin-based vector pLSU2 in promoting a 10-fold increase in fresh weight yield of kanamycin-resistant calli after A. tumefaciens-mediated transformation of tobacco leaf discs. Using the Gateway Technology we introduced the plant-expressible GUSgene to the T-DNA of binary Ti vector pLSU12. Expression of the β-glucuronidase enzyme activity was demonstrated by histochemical staining of the GUS activity in transformed tobacco leaf discs.

show abstract

“…Organisms that have been genetically modified include micro-organisms such as bacteria and yeast, insects, plants, fish, and mammals [1][2][3]. GMOs are the principal source of genetically modified foods, and are thus widely used in scientific research and to produce goods other than food.…”

Section: Introductionmentioning

confidence: 99%

Physiological and Biochemical Significance of Genetically Modified Foods: An Overview

Mishra¹

2013

TONUTRAJ

View full text Add to dashboard Cite

Biotechnological discoveries and inventions have been observed to improve food production qualitatively as well as quantitatively. In certain specific cases, the improvement in the quality and nutrition of foods by altering their composition were also monitored. However, the practice of biotechnology has also upraised concerns about its potential risks to the environment as well as human being. Genetic Engineering provides resources to host genes into plants via mechanisms, different in some respects from classical breeding. A number of genetically engineered variety of foods have been developed, which have become important nutraceuticals; most notably canola, cotton, maize and soybean, were developed employing this modern technology, and at present the traits introduced are herbicide and/or pest tolerance. Gene technology leads to increase the production in plants, as well as the elevation of resistance to pests, viruses, frost, etc. Gene transfer technology is employed to alter the physical and chemical composition with nutraceutical worth. The present review article is the compilation of various physiological and biochemical studies reflecting both positive and negative ecological concerns of genetically modified foods.

show abstract

T-DNA Binary Vectors and Systems

Cited by 198 publications

References 103 publications

The Small Subunit of Snapdragon Geranyl Diphosphate Synthase Modifies the Chain Length Specificity of Tobacco Geranylgeranyl Diphosphate Synthase in Planta

The Small Subunit of Snapdragon Geranyl Diphosphate Synthase Modifies the Chain Length Specificity of Tobacco Geranylgeranyl Diphosphate Synthase in Planta

Tetracycline-Based Binary Ti Vectors pLSU with Efficient Cloning by the Gateway Technology for <i>Agrobacterium tumefaciens</i>-Mediated Transformation of Higher Plants

Physiological and Biochemical Significance of Genetically Modified Foods: An Overview

Contact Info

Product

Resources

About

T-DNA Binary Vectors and Systems

Cited by 198 publications

References 103 publications

The Small Subunit of Snapdragon Geranyl Diphosphate Synthase Modifies the Chain Length Specificity of Tobacco Geranylgeranyl Diphosphate Synthase in Planta

The Small Subunit of Snapdragon Geranyl Diphosphate Synthase Modifies the Chain Length Specificity of Tobacco Geranylgeranyl Diphosphate Synthase in Planta

Tetracycline-Based Binary Ti Vectors pLSU with Efficient Cloning by the Gateway Technology for &lt;i&gt;Agrobacterium tumefaciens&lt;/i&gt;-Mediated Transformation of Higher Plants

Physiological and Biochemical Significance of Genetically Modified Foods: An Overview

Contact Info

Product

Resources

About

Tetracycline-Based Binary Ti Vectors pLSU with Efficient Cloning by the Gateway Technology for <i>Agrobacterium tumefaciens</i>-Mediated Transformation of Higher Plants