Differential Display as a Method of Detecting Gene Expression Differences in Obese and Lean Mice
Johnson, Kelli R.
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This paper shows that gene expression differences can be detected using the differential display technique developed by Peng Liang and Arthur Pardee. The principle behind differential display is to amplify the 3' terminal portions of mRNAs on a DNA sequencing gel. Downstream primers designed to bind to the 5' end of a poly A tail are used for the reverse transcription followed by PCR amplification with additional arbitrary upstream primers. Visualization of mRNA is done by denaturing polyacrylamide electrophoresis. We found that five samples of hypothalamus from transgenic obese (+db/+db) and lean (+m/+m) mice displayed gene expression differences. These differentially expressed cDNA samples were then recovered and used to clone their cDNAs. The results of the cloning is in progress, however, it is possible that one of the subclones is the db gene.