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General Information

Document type
  • Peer-reviewed journal article
GE organism
  • soybean
GE trait
  • quality
Country
  • USA

Results

Efficacy
  • positive effect

Stacking multiple transgenes at a selected genomic site via repeated recombinase-mediated DNA cassette exchanges Open Access

Li, Z; Moon, BP; Xing, A; Liu, ZB; McCardell, RP; Damude, HG; Falco, SC
Plant Physiology. 2010 October. 154(2):622-631

Link to full text (open access, freely available)

PMID: 20720171 DOI: ​10.​1104/​pp.​110.​160093 ISSN: 1532-2548

Abstract

Recombinase-mediated DNA cassette exchange (RMCE) has been successfully used to insert transgenes at previously characterized genomic sites in plants. Following the same strategy, groups of transgenes can be stacked to the same site through multiple rounds of RMCE. A gene-silencing cassette, designed to simultaneously silence soybean (Glycine max) genes fatty acid ω-6 desaturase 2 (FAD2) and acyl-acyl carrier protein thioesterase 2 (FATB) to improve oleic acid content, was first inserted by RMCE at a precharacterized genomic site in soybean. Selected transgenic events were subsequently retransformed with the second DNA construct containing a Yarrowia lipolytica diacylglycerol acyltransferase gene (DGAT1) to increase oil content by the enhancement of triacylglycerol biosynthesis and three other genes, a Corynebacterium glutamicum dihydrodipicolinate synthetase gene (DHPS), a barley (Hordeum vulgare) high-lysine protein gene (BHL8), and a truncated soybean cysteine synthase gene (CGS), to improve the contents of the essential amino acids lysine and methionine. Molecular characterization confirmed that the second RMCE successfully stacked the four overexpression cassettes to the previously integrated FAD2-FATB gene-silencing cassette. Phenotypic analyses indicated that all the transgenes expressed expected phenotypes.

Keywords

transgene, stacking, soybean, cassette

Funding

Funding source
  • DuPont Pioneer
Funding country
  • United States
Funding type
  • industry: same

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Cite this study

MLA

Li, Z, BP Moon, A Xing, ZB Liu, RP McCardell, HG Damude, SC Falco. "Stacking multiple transgenes at a selected genomic site via repeated recombinase-mediated DNA cassette exchanges." Plant Physiology 154.2 (2010): 622-631 . Web. 27 Jul. 2017.

APA

Li, Z., Moon, BP., Xing, A., Liu, ZB., McCardell, RP., Damude, HG., & Falco, SC. (2010). Stacking multiple transgenes at a selected genomic site via repeated recombinase-mediated DNA cassette exchanges. Plant Physiology, 154(2), 622-631 . doi:​10.​1104/​pp.​110.​160093

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