FY2010 - Oak Ridge National Laboratory

Seed Money Fund—
Biosciences Division
Salmonella cells were grown overnight to stationary phase. The bacteria were killed by exposure to 70%
ethanol for 20 min. After 12 or 24 hours, both viable and nonviable cells were subject to Trizol treatment,
followed by the use of the RNeasy Mini Kit (Qiagen) to extract RNA. Remnants of DNA present in the
extracted RNA will be digested by DNase I. Subsequently, 0.3 μg RNA will be reversely transcribed into
cDNA and used as template for PCR amplification. Multiplex PCR using a 0.8 μM primer mixture
composed of all the selected primers was carried out. In control experiments, the presence of traces of
DNA was examined by using the RNA sample as template in PCR.
A combination of a set of primers (ttrA forward, GTGGGCGGTACAATATTTCTTTT; reverse,
TCACGAATAATAATCAGTAGCGC; iroB forward, TGCGTATTCTGTTTGTCGGTCC; reverse,
TACGTTCCCACCATTCTTCCC; ttr forward, ACTGCCGATAAATGCACGTT; reverse,
CTTTTTTCCGCCAGTGAAGA; invA forward, CGCTCTTTCGTCTGGCATTATC; reverse,
CCGCCAATAAAGTTCACAAAG; invA forward, GTGAAATTATCGCCACGTTCG; reverse,
CATCGCACCGTCAAAGGAA; rpoD forward, CCGATGAAGATGCGGAAGAAGC; reverse,
CAAACGAGCCTCTTCAGCCT) was able to generate bands of expected sizes. The PCR condition was
94 degree 2 min (94 degree 30 sec, 55 degree 30 sec, 72 degree 1 min) for 35 cycles, 72 degree 10 min.
Clear RT-PCR bands were detected for viable cells but not for cells prepared at 12 or 24 hours after
ethanol treatment, suggesting that these primers were effective for differentiating viable from nonviable
cells under the given condition.
Multiplex PCR was tested for detecting Salmonella in the presence of food matrices—minced meat or
egg. However, to date no bands were successfully generated. Nanodrop reading indicated that the
extracted RNA was of poor quality, so it is necessary to modify the existing DNA extraction protocol to
remove possible inhibitory components in the food matrices that prevent the success of reverse
transcription or PCR.
05879
White Light Produced by a Scalable Biosynthesized Zinc Gallate
Mixture
Ji-Won Moon, Tommy J. Phelps, Chad E. Duty, Gerald E. Jellison, Jr., and Lonnie J. Love
Project Description
We are pursuing a new approach to generating white light using scalable and economical microbial
production of zinc gallate phosphors which can emit red, green, and blue (RGB) colors for use in energyefficient
solid state lighting (SSL). Improvement over current SSL technology requires advances in
several areas: (1) improved reactions to produce appropriate phosphors, (2) less complicated fabrication,
(3) better control over stoichiometry during mass production, and (4) ways to eliminate or cope with
unexpected secondary phases. NanoFermentation, which employs microbes to produce high-quality
nanoparticles, can provide (1) consistent, nanoscale particle size without energy-intensive milling;
(2) ease of stoichiometric control; (3) reproducibility; and (4) a low-temperature, scalable, and
economical process.
In this project, we explore the unprecedented use of NanoFermentation to produce controllably doped
zinc-gallate nanoparticles that emit red, green and blue light, and then combine these single-color
particles to produce white light. This research is framed during FY 2010–2011, starting in July 2010, and
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