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FLP-mediated conditional loss of an essential gene to facilitate complementation assays

Description: Commonly, when it is desirable to replace an essential gene with an allelic series of mutated genes, or genes with altered expression patterns, the complementing constructs are introduced into heterozygous plants, followed by the selection of homozygous null segregants. To overcome this laborious and time-consuming step, the newly developed two-component system utilizes a site-specific recombinase to excise a wild-type copy of the gene of interest from transformed tissues. In the first component (the first vector), a wild-type version of the gene is placed between target sequences recognized by FLP recombinase from the yeast 2 μm plasmid. This construct is transformed into a plant heterozygous for a null mutation at the endogenous locus, and progeny plants carrying the excisable complementing gene and segregating homozygous knockout at the endogenous locus are selected. The second component (the second vector) carries the experimental gene along with the FLP gene. When this construct is introduced, FLP recombinase excises the complementing gene, leaving the experimental gene as the only functional copy. The FLP gene is driven by an egg apparatus specific enhancer (EASE) to ensure excision of the complementing cDNA in the egg cell and zygote following floral-dip transformation. The utility of this system is being tested using various experimental derivatives of the essential sucrose-proton symporter, AtSUC2, which is required for photoassimilate transport.
Date: December 2007
Creator: Ganesan, Savita
Partner: UNT Libraries

A cytochrome P450 monooxygenase commonly used for negative selection in transgenic plants causes growth anomalies by disrupting brassinosteroid signaling

Description: Article on a cytochrome P450 monooxygenase commonly used for negative selection in transgenic plants causing growth anomalies by disrupting brassinosteroid signaling.
Date: April 15, 2011
Creator: Dasgupta, Kasturi; Ganesan, Savita; Manivasagam, Sindhu & Ayre, Brian G.
Partner: UNT College of Arts and Sciences

Effective carbon partitioning driven by exotic phloem-specific regulatory elements fused to the Arabidopsis thaliana AtSUC2 sucrose-proton symporter gene

Description: This article discusses the effect and overall importance of the Arabidopsis thaliana AtSUC2 sucrose-proton symporter gene on carbon partitioning and plant growth.
Date: January 20, 2009
Creator: Srivastava, Avinash C.; Ganesan, Savita; Ismail, Ihab O. & Ayre, Brian G.
Partner: UNT College of Arts and Sciences