Application of fluorescently labeled poly(dU) for gene expression profiling on cDNA microarrays.
AuthorsMeiyanto E, Mineno J, Ishida N, Takeya T
JournalBiotechniques
PubMed ID11515377
'The membrane filter hybridization technique has been widely used for gene expression profiling. The preparation of sensitive and reliable probes is critical for quantitative analysis in this technique. We report a method in which fluorescently labeled poly(dU) is used to detect poly(A)-containing mRNA that hybridizes to specific gene targets. The ... More
Single-molecule analysis of 1D diffusion and transcription elongation of T7 RNA polymerase along individual stretched DNA molecules.
AuthorsKim JH, Larson RG,
JournalNucleic Acids Res
PubMed ID17526520
Using total internal reflection fluorescence microscopy, we directly visualize in real-time, the 1D Brownian motion and transcription elongation of T7 RNA polymerase along aligned DNA molecules bound to substrates by molecular combing. We fluorescently label T7 RNA polymerase with antibodies and use flow to convect them orthogonally to the DNA ... More
Drosophila Squid/hnRNP helps Dynein switch from a gurken mRNA transport motor to an ultrastructural static anchor in sponge bodies.
AuthorsDelanoue R, Herpers B, Soetaert J, Davis I, Rabouille C,
JournalDev Cell
PubMed ID17925228
In Drosophila oocytes, dorso-anterior transport of gurken mRNA requires both the Dynein motor and the heterogeneous nuclear ribonucleoprotein (hnRNP) Squid. We show that gurken transcripts are transported directly on microtubules by Dynein in nonmembranous electron-dense transport particles that also contain Squid and the transport cofactors Egalitarian and Bicaudal-D. At its ... More
In vivo analysis of Drosophila bicoid mRNA localization reveals a novel microtubule-dependent axis specification pathway.
AuthorsCha BJ, Koppetsch BS, Theurkauf WE
JournalCell
PubMed ID11461700
Drosophila bicoid mRNA is synthesized in the nurse cells and transported to the oocyte where microtubules and Exuperantia protein mediate localization to the anterior pole. Fluorescent bicoid mRNA injected into the oocyte displays nonpolar microtubule-dependent transport to the closest cortical surface, and the oocyte microtubule cytoskeleton lacks clear axial asymmetry. ... More
Sequence-specific fluorescent labeling of double-stranded DNA observed at the single molecule level.
AuthorsGéron-Landre B, Roulon T, Desbiolles P, Escudé C
JournalNucleic Acids Res
PubMed ID14530458
Fluorescent labeling of a short sequence of double-stranded DNA (dsDNA) was achieved by ligating a labeled dsDNA fragment to a stem-loop triplex forming oligonucleotide (TFO). After the TFO has wound around the target sequence by ligand-induced triple helix formation, its extremities hybridize to each other, leaving a dangling single-stranded sequence, ... More
Steady-state dynamics of Cajal body components in the Xenopus germinal vesicle.
AuthorsHandwerger KE, Murphy C, Gall JG
JournalJ Cell Biol
PubMed ID12591912
Cajal bodies (CBs) are evolutionarily conserved nuclear organelles that contain many factors involved in the transcription and processing of RNA. It has been suggested that macromolecular complexes preassemble or undergo maturation within CBs before they function elsewhere in the nucleus. Most such models of CB function predict a continuous flow ... More