Biosynthetic Labelling of RNA with Uracil Phosphoribosyltransferase Allows Cell Specific Microarray Analysis of mRNA Synthesis and Decay  

Oleh:

Cleary, Michael D. ; Meiering, Christopher D. ; Jan, Eric ; Guymon, Rebecca ; Boothroyd, John C.

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Nature Biotechnology: The Science and Business of Biotechnology vol. 23 no. 2 (Feb. 2005), page 232-237.
Topik: rna; biosynthetic; RNA; microarray; synthesis and decay

Ketersediaan

Perpustakaan Pusat (Semanggi) Nomor Panggil: NN9.2 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

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Isi artikel

Standard microarrays measure mRNA abundance, not mRNA synthesis, and therefore cannot identify the mechanisms that regulate gene expression. We have developed a method to overcome this limitation by using the salvage enzyme uracil phosphoribosyltransferase (UPRT) from the protozoan toxoplasma gondii. T. gondii UPRT has been well characterized because of its application in monitoring parasite growth, mammals lack this enzyme activity and thus only the parasite incorporates H-uracil into its nucleic acids. In this study we used RNA labelling by UPRT to determine expression during T. gondii asexual development. We also used this approach to specifically label parasite RNA during a mouse infection and to incorporate this substituted uridines into the RNA of human cells engineered to express T. gondii UPRT, indicating that engineered UPRT expression will allow cell specific analysis of gene expression will allow cell specific analysis of gene expression in organisms other than T. gondii.

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