The Experts below are selected from a list of 27 Experts worldwide ranked by ideXlab platform
Bernard Moss - One of the best experts on this subject based on the ideXlab platform.
-
purification and use of vaccinia Virus Messenger RNA capping enzyme
Methods in Enzymology, 1990Co-Authors: Stewart Shuman, Bernard MossAbstract:Publisher Summary This chapter provides an overview of the properties of the vaccinia capping enzyme, detailed protocols for enzyme assay and purification, and examples of how the purified enzyme may be of use to biochemists and molecular biologists. As the vaccinia capping enzyme exists as a complex of RNA triphosphatase, RNA guanylyltransferase, and RNA (guanine-7)-methyltransferase activities, any one of the three functions may be used to monitor enzyme activity during purification. Polyadenylic acid with a 5′-triphosphate terminus is generally used as the cap acceptor for assay purposes, and it may be synthesized by Escherichia coli RNA polymerase using the following protocol. Electrophoresis must be carried out until the dye front is well out of the bottom of the gel. The samples may be precipitated with TCA after denaturation with SDS and the acid-insoluble radioactivity determined by collection on glass fiber filters. Vaccinia is stable at ambient temperature even when dried, and parenteral inoculation and droplet or aerosol exposure of mucous membranes or broken skin with infectious Virus are the primary hazards to laboratory personnel.
Stewart Shuman - One of the best experts on this subject based on the ideXlab platform.
-
purification and use of vaccinia Virus Messenger RNA capping enzyme
Methods in Enzymology, 1990Co-Authors: Stewart Shuman, Bernard MossAbstract:Publisher Summary This chapter provides an overview of the properties of the vaccinia capping enzyme, detailed protocols for enzyme assay and purification, and examples of how the purified enzyme may be of use to biochemists and molecular biologists. As the vaccinia capping enzyme exists as a complex of RNA triphosphatase, RNA guanylyltransferase, and RNA (guanine-7)-methyltransferase activities, any one of the three functions may be used to monitor enzyme activity during purification. Polyadenylic acid with a 5′-triphosphate terminus is generally used as the cap acceptor for assay purposes, and it may be synthesized by Escherichia coli RNA polymerase using the following protocol. Electrophoresis must be carried out until the dye front is well out of the bottom of the gel. The samples may be precipitated with TCA after denaturation with SDS and the acid-insoluble radioactivity determined by collection on glass fiber filters. Vaccinia is stable at ambient temperature even when dried, and parenteral inoculation and droplet or aerosol exposure of mucous membranes or broken skin with infectious Virus are the primary hazards to laboratory personnel.
G Delsol - One of the best experts on this subject based on the ideXlab platform.
-
detection of epstein barr Virus Messenger RNA in reed sternberg cells of hodgkin s disease by in situ hybridization with biotinylated probes on specially processed modified acetone methyl benzoate xylene modamex sections see comments
Blood, 1991Co-Authors: Pierre Brousset, S Chittal, D Schlaifer, J Icart, Catherine Payen, Francoise Rigalhuguet, J J Voigt, G DelsolAbstract:Microscopic intracellular detection of Epstein-Barr Virus (EBV) Messenger RNA in Reed-Sternberg cells of Hodgkin9s disease (HD) was possible by in situ hybridization, in tissue sections prepared by a method termed modified acetone methyl benzoate xylene (ModAMeX). The ModAMeX method was initially developed for simultaneous optimal preservation of leucocyte differentiation antigens and morphology. Two biotinylated DNA probes, corresponding to the same BamHI-W (inteRNAl repeat) of the EBV genome were used. EBV mRNA was detected in neoplastic cells in 16 of 54 (30%) lymph node biopsy specimens from usual subtypes of HD (lymphocyte predominance, 0/5; nodular sclerosis, 4/22; mixed cellularity, 12/26; unclassified, 0/1). EBV mRNA was also detected in the lymph node biopsy of 1 additional human immunodeficiency Virus (HIV)-related case of HD (mixed cellularity) and in 2 of 4 cases of B-cell lymphomas occurring in patients with acquired immunodeficiency syndrome (AIDS). In other non-Hodgkin9s lymphomas, EBV mRNA was detected in only 1 of 41 cases. Cases of HD positive for EBV mRNA were immunostained by CD30 and CD15 antibodies. The hybridization signals were exclusively restricted to Reed-Sternberg cells and variants. When analyzed retrospectively, no statistically significant correlation emerged between hybridization findings, EBV serology, or disease outcome over the 3 years of the availability of ModAMeX technique. The findings support the contention of a direct role of EBV in the pathogenesis of HD, at least in some cases.
Pierre Brousset - One of the best experts on this subject based on the ideXlab platform.
-
detection of epstein barr Virus Messenger RNA in reed sternberg cells of hodgkin s disease by in situ hybridization with biotinylated probes on specially processed modified acetone methyl benzoate xylene modamex sections see comments
Blood, 1991Co-Authors: Pierre Brousset, S Chittal, D Schlaifer, J Icart, Catherine Payen, Francoise Rigalhuguet, J J Voigt, G DelsolAbstract:Microscopic intracellular detection of Epstein-Barr Virus (EBV) Messenger RNA in Reed-Sternberg cells of Hodgkin9s disease (HD) was possible by in situ hybridization, in tissue sections prepared by a method termed modified acetone methyl benzoate xylene (ModAMeX). The ModAMeX method was initially developed for simultaneous optimal preservation of leucocyte differentiation antigens and morphology. Two biotinylated DNA probes, corresponding to the same BamHI-W (inteRNAl repeat) of the EBV genome were used. EBV mRNA was detected in neoplastic cells in 16 of 54 (30%) lymph node biopsy specimens from usual subtypes of HD (lymphocyte predominance, 0/5; nodular sclerosis, 4/22; mixed cellularity, 12/26; unclassified, 0/1). EBV mRNA was also detected in the lymph node biopsy of 1 additional human immunodeficiency Virus (HIV)-related case of HD (mixed cellularity) and in 2 of 4 cases of B-cell lymphomas occurring in patients with acquired immunodeficiency syndrome (AIDS). In other non-Hodgkin9s lymphomas, EBV mRNA was detected in only 1 of 41 cases. Cases of HD positive for EBV mRNA were immunostained by CD30 and CD15 antibodies. The hybridization signals were exclusively restricted to Reed-Sternberg cells and variants. When analyzed retrospectively, no statistically significant correlation emerged between hybridization findings, EBV serology, or disease outcome over the 3 years of the availability of ModAMeX technique. The findings support the contention of a direct role of EBV in the pathogenesis of HD, at least in some cases.
Paul S Miller - One of the best experts on this subject based on the ideXlab platform.
-
interactions of psoralen derivatized oligodeoxyribonucleoside methylphosphonates with vesicular stomatitis Virus Messenger RNA
Antisense research and development, 1994Co-Authors: Joel T Levis, Paul S MillerAbstract:The ability of oligonucleotides to interact selectively with their targets is an important consideration in the design of antisense oligonucleotides. This is especially important in the case of antisense oligomers, such as psoralen-derivatized oligomers, which can irreversibly bind to their targets. We have studied the interactions of a series of psoralen-derivatized antisense oligonucleoside methylphosphonates with the mRNAs of vesicular stomatitis Virus (VSV), mRNAs that have a high degree of sequence homology. Cross-linking reactions were carried out under conditions of low ionic strength in order to reduce mRNA secondary structure. A 12-mer, whose sequence was complementary to VSV M -mRNA and partially complementary to sequences found in N, NS, and G mRNA cross-linked extensively to N-message. On the other hand, 16-mers whose sequences were uniquely complementary to binding sites on N- or M-mRNA specifically and efficiently cross-linked to their targeted mRNAs over the temperature range 0° to 37°C. A ...
