3 resultados para RECOGNITION SITES

em Deakin Research Online - Australia


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Recognition of poly(A) sites in yeast pre-mRNAs is poorly understood. Employing an in vitro cleavage system with cleavage and polyadenylation factor (CPF) and cleavage factor IA we show that the efficiency and positioning elements are dispensable for poly(A)-site recognition within a short CYC1 substrate in vitro. Instead, U-rich elements immediately upstream and downstream of the poly(A) site mediate cleavage-site recognition within CYC1 and ADH1 pre-mRNAs. These elements act in concert with the poly(A) site to produce multiple recognition sites for the processing machinery, since combinations of mutations within these elements were most effective in cleavage inhibition. Intriguingly, introduction of a U-rich element downstream of the GAL7 poly(A) site strongly enhanced cleavage, underscoring the importance of downstream sequences in general. RNA- binding analyses demonstrate that cleavage depends on the recognition of the poly(A)-site region by CPF. Consistent with in vitro results, mutation of sequences upstream and downstream of the poly(A) site affected 3'-end formation in vivo. A model for yeast pre-mRNA cleavage-site recognition outlines an unanticipated high conservation of yeast and mammalian 3'-end processing mechanisms.

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In total, 18 novel norbornene and [n]polynorbornane based anion receptors were synthesised and evaluated. The host:guest complexes examined were influenced by the position and number of available recognition sites within each binding cleft. It was concluded that [n]polynorbornane scaffolds are ideal for studying the fundamental concepts of supramolecular chemistry.

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Two methods for attaching DNA to oxidized single-walled carbon nanotubes either in organic solvent or aqueous solution are described. The sites of DNA attachment to the nanotubes have been verified by binding gold nanoparticles modified with DNA of complementary sequence to the DNA-modified nanotubes, and imaging with TEM. The gold nanoparticles appear on the tips of the nanotubes, and at isolated positions (defects) on the sidewalls. The methods provide versatility for the modification of nanotubes with DNA for their directed assembly, or for their composites with gold nanoparticles, into nanoscale devices.