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Gel electrophoresis
About: Gel electrophoresis is a research topic. Over the lifetime, 26026 publications have been published within this topic receiving 1113565 citations.
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TL;DR: To identify the viral core protein that catalyzes the initiation of transcription via the incorporation of a guanosine residue onto primer fragments, irradiated transcription reactions carried out by viral cores in the presence of [alpha-P]GTP as the only ribonucleoside triphosphate with an unlabeled primer RNA suggested that the viral P1 protein catalyzes this incorporation and hence initiates transcription.
Abstract: Purified influenza viral cores catalyze the entire process of viral RNA transcription, which includes the endonucleolytic cleavage of heterologous RNAs containing cap 1 (m(7)GpppNm) structures to generate capped primers 10-13 nucleotides long, the initiation of transcription via the incorporation of a guanosine residue onto the primers, and elongation of the viral mRNAs [Plotch, S. J., Bouloy, M., Ulmanen, L & Krug, R. M. (1980) Cell 23, 847-858]. To identify which viral core protein (nucleocapsid protein, P1, P2, or P3) recognizes the cap 1 structure on the RNA primer, we irradiated (UV) endonuclease reactions carried out by viral cores in the absence of ribonucleoside triphosphates, with a primer RNA labeled in its cap 1 structure with (32)P. The labeled cap was crosslinked to a protein that had a mobility similar to that of the P3 protein, the smaller of the two basic P proteins, in both one- and two-dimensional gel electrophoresis. This strongly suggests that this crosslinked protein is the viral P3 protein. Competition experiments with unlabeled RNAs containing or lacking a cap 1 structure established that this protein recognizes the cap 1 structure on RNAs. This protein remained associated with the cap throughout the transcription reaction, even after the viral mRNA molecules were elongated. To identify the viral core protein that catalyzes the initiation of transcription via the incorporation of a guanosine residue onto primer fragments, we irradiated transcription reactions carried out by viral cores in the presence of [alpha-(32)P]GTP as the only ribonucleoside triphosphate with an unlabeled primer RNA. A labeled guanosine residue was crosslinked to a protein that had a mobility similar to that of the P1 protein, the larger of the two basic P proteins, in both one-and two-dimensional gel electrophoresis. The transcription reaction conditions required to bring this protein in close association with a labeled guanosine residue so that crosslinking could occur indicated that this association most likely occurred coincident with the guanosine residue's being incorporated onto the primer. These results suggest that the viral P1 protein catalyzes this incorporation and hence initiates transcription.
254 citations
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TL;DR: It is demonstrated that revertants of tem- perature-sensitive benA (p-tubulin) mutations in Aspergillus nidulans can be used to identify pro- teins which interact with /3-tubulins and the potential usefulness of the benA sys- tem function for analyzing tubulin structure and function is extended.
253 citations
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TL;DR: A two-dimensional electrophoresis system for analysis of ribosomal proteins with several advantages over previous systems is described, and is relatively simple and inexpensive to construct and use.
253 citations
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TL;DR: A chromatographically and immunologically identical collagenase inhibitor was partially purified from human serum, suggesting the possibility that the fibroblast-derived inhibitor and the previously reported serum beta 1-anticollagenase are similar, if not identical, proteins.
252 citations
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15 Oct 1990
TL;DR: In this article, a process for the electrophoretic analysis of DNA fragments produced in DNA sequencing operations is described, wherein chromophores or fluorophores are used to tag the DNA fragments and permit the detection and characterization of the fragments as they are resolved by electrophoresis through a gel.
Abstract: A process for the electrophoretic analysis of DNA fragments produced in DNA sequencing operations wherein chromophores or fluorophores are used to tag the DNA fragments produced by the sequencing chemistry and permit the detection and characterization of the fragments as they are resolved by electrophoresis through a gel. Preferably four different fragment sets are tagged with the fluorophores fluorescein, Texas Red, tetramethyl rhodamine, and 7-nitrobenzofurazan. A system for the electrophoretic analysis of DNA fragments produced in DNA sequencing operations comprising: a source of chromophore or fluorescent tagged DNA fragments; a zone for contacting an electrophoresis gel; means for introducing said tagged DNA fragments to said zone; and photometric means for monitoring said tagged DNA fragments as they move through said gel.
252 citations