Saint Louis, United States

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    ABSTRACT: Key message: Agrobacterium tumefaciens mediates high frequency of germline transformation of cotton meristem explants. The meristem transformation system we developed is rapid, high throughput and genotype-flexible. We have developed a high throughput cotton transformation system based on direct Agrobacterium inoculation of mechanically isolated meristem explants of cotton (Gossypium hirsutum L.). The explants were inoculated with a disarmed A. tumefaciens strain, AB33 harboring a 2 T-DNA binary vector pMON114908. This vector contained a gene of interest, an intron-disrupted β-glucuronidase gene in one T-DNA, and a selectable marker gene, aadA in the other T-DNA. Critical factors, such as method of co-culture, culture temperature during selection, composition of selection medium, and selection scheme were found to influence transformation frequency. The cycle time from initial inoculation to the transplanting of transgenic plants to soil was 7-8 weeks. Stable integration of transgenes and their transmission to progeny were confirmed by molecular and genetic analyses. Transgenes segregated in the expected Mendelian fashion in the T1 generation for most of the transgenic events. It was possible to recover marker-free events in the T1 generation when utilizing a binary vector that contained the selectable marker and gene of interest expression cassettes on independent T-DNAs. The procedure presented here has been used to regenerate thousands of independent transgenic events from multiple varieties with numerous constructs, and we believe it represents a major step forward in cotton transformation technology.
    Plant Cell Reports 10/2013; 33(1). DOI:10.1007/s00299-013-1519-x
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    ABSTRACT: Myristoylation is a lipid modification conserved among eukaryotes and involves the addition of a 14-carbon myristoyl moiety to a glycine at the N-terminus of cargo proteins. Since not every protein with an N-terminal glycine is myristoylated, experimental verification is necessary to determine which proteins are indeed myristoylated. Here we describe an in vitro myristoylation assay for the Arabidopsis heterotrimeric G protein alpha subunit, GPA1, as well as the Arabidopsis SALT OVERLY SENSITIVE3. This method can be easily adopted to other proteins of interest.
    Methods in molecular biology (Clifton, N.J.) 08/2013; 1043:135-9. DOI:10.1007/978-1-62703-532-3_14
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    ABSTRACT: "Field-evolved resistance" is defined as a "genetically based decrease in susceptibility of a population to a toxin caused by exposure to the toxin in the field." The key component of "field-evolved" resistance is that it does confer decreased susceptibility to an insecticide in the field. Another key component is that the decrease in susceptibility to the insecticide is because of previous exposure of the target insect to the toxin in the field. Several studies have reported field-evolved resistance to crops engineered to express proteins from the bacterium, Bacillus thuringiensis (Bt). However, there has not been a consistent standard in the application of the definition of field-evolved resistance for Bt crops. The inconsistency in applying the definition arises from differences in the methods used to detect resistance, the ecology of the interaction between the pest and the Bt crop, and the effective dose the pest encounters while feeding on the Bt crop. Using case studies of reported resistance to Bt crops, it is demonstrated resistance does not come in a single form, and that in most cases, resistance can still be managed.
    Journal of Economic Entomology 08/2013; 106(4):1525-34. DOI:10.1603/EC13103


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    Saint Louis, United States
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Top publications last week by reads

Biotechnology(Faisalabad) 08/1987; 5(8). DOI:10.1038/nbt0887-807
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Physiologia Plantarum 04/2006; 90(4):791 - 800. DOI:10.1111/j.1399-3054.1994.tb02539.x
38 Reads

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