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- 저자 : Karl Pfeifer (검색결과 2 건)
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김재영,윤화영,Karl Pfeifer,은복기 한국통합생물학회 2019 Animal cells and systems Vol.23 No.4
Igf2, H19, and Nctc1 are linked co-regulated genes on distal mouse chromosome 7. This locus is an important model both for studying mechanisms of monoallelic expression and for elucidating the role of cis-regulatory elements – enhancers and insulators – in organizing chromatin and gene expression across a large domain. In this study we characterize regulated expression of the Igf2 antisense transcript (Igf2as) in primary muscle cells. We demonstrate that Igf2as is imprinted (expressed only from the paternal chromosome). We also show that Igf2as expression during differentiation follows the same patterns as Igf2 and H19. Moreover, this expression is dependent upon the same shared enhancer element. Thus, our work shows that the imprinted cluster includes Igf2as in addition to H19, Igf2, and Nctc1.
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Accurate Measurement of the Relative Abundance of Different DNA Species in Complex DNA Mixtures
Jeong, Sangkyun,Yu, Hyunjoo,Pfeifer, Karl Oxford University Press 2012 DNA research Vol.19 No.3
<P>A molecular tool that can compare the abundances of different DNA sequences is necessary for comparing intergenic or interspecific gene expression. We devised and verified such a tool using a quantitative competitive polymerase chain reaction approach. For this approach, we adapted a competitor array, an artificially made plasmid DNA in which all the competitor templates for the target DNAs are arranged with a defined ratio, and melting analysis for allele quantitation for accurate quantitation of the fractional ratios of competitively amplified DNAs. Assays on two sets of DNA mixtures with explicitly known compositional structures of the test sequences were performed. The resultant average relative errors of 0.059 and 0.021 emphasize the highly accurate nature of this method. Furthermore, the method's capability of obtaining biological data is demonstrated by the fact that it can illustrate the tissue-specific quantitative expression signatures of the three housekeeping genes <I>G6pdx</I>, <I>Ubc</I>, and <I>Rps27</I> by using the forms of the relative abundances of their transcripts, and the differential preferences of <I>Igf2</I> enhancers for each of the multiple <I>Igf2</I> promoters for the transcription.</P>
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