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We previously constructed a recombinant monoclonal antibody (rec-MAb 63P4) that detects immediate-early protein IE63 encoded by varicella-zoster virus (VZV) in the cytoplasm of productively infected cells. Here, we used ORF63 truncation mutants to map the rec-MAb 63P4 binding epitope to amino acids 141 to 150 of VZV IE63, a region not shared with other widely used anti-IE63 antibodies, and found that the recombinant antibody does not bind to the simian IE63 counterpart.  相似文献   
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We have evaluated codon usage bias in Drosophila histone genes and have obtained the nucleotide sequence of a 5,161-bp D. hydei histone gene repeat unit. This repeat contains genes for all five histone proteins (H1, H2a, H2b, H3, and H4) and differs from the previously reported one by a second EcoRI site. These D. hydei repeats have been aligned to each other and to the 5.0-kb (i.e., long) and 4.8-kb (i.e., short) histone repeat types from D. melanogaster. In each species, base composition at synonymous sites is similar to the average genomic composition and approaches that in the small intergenic spacers of the histone gene repeats. Accumulation of synonymous changes at synonymous sites after the species diverged is quite high. Both of these features are consistent with the relatively low codon usage bias observed in these genes when compared with other Drosophila genes. Thus, the generalization that abundantly expressed genes in Drosophila have high codon bias and low rates of silent substitution does not hold for the histone genes.   相似文献   
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The presence of increased IgG in the brains of humans with infectious and inflammatory CNS diseases of unknown etiology such as multiple sclerosis may be a clue to the cause of disease. For example, the intrathecally synthesized oligoclonal bands in diseases such as subacute sclerosing panencephalitis (SSPE) or cryptococcal meningitis have been shown to represent Ab directed against the causative agents, measles virus (MV), or Cryptococcus neoformans, respectively. Using SSPE as a model system, we developed a strategy to identify the antigenic targets of the intrathecal disease-relevant IgG in chronic human inflammatory and demyelinating diseases of the CNS. Libraries of cDNA Ags were displayed on the surface of T7Select bacteriophage and biopanned on IgG extracted from the brain of an SSPE patient, or on a monospecific recombinant Fab identified from SSPE brain. After three or six rounds of biopanning on either Ab, positive phage-displayed Ags reacting with IgG were enriched to 35-77% of all panned clones. Sequence analysis of the positive clones identified fragments of the nucleocapsid protein of MV, the cause of SSPE. The sensitivity of the system was determined by diluting the positive clones from this SSPE phage-displayed library at a ratio of 10(-6) into another phage-displayed library that did not contain any detectable MV Ags; after six rounds of panning, the positive clones comprised 34% of all phage and were also shown to be MV nucleocapsid specific. This strategy will be useful to identify potentially rare Ags in diseases of unknown cause.  相似文献   
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Varicella-zoster virus (VZV) is a ubiquitous, highly cell-associated, and exclusively human neurotropic alphaherpesvirus. VZV infection is initiated by membrane fusion, an event dependent in part on VZV glycoproteins gH and gL. Consistent with its location on the virus envelope, the gH/gL complex is a target of neutralizing antibodies produced after virus infection. One week after immunizing a 59-year-old VZV-seropositive man with Zostavax, we sorted his circulating blood plasma blasts and amplified expressed immunoglobulin variable domain sequences by single-cell PCR. Sequence analysis identified two plasma blast clones, one of which was used to construct a recombinant monoclonal antibody (rec-RC IgG). The rec-RC IgG colocalized with VZV gE on the membranes of VZV-infected cells and neutralized VZV infection in tissue culture. Mass spectrometric analysis of proteins immunoprecipitated by rec-RC IgG identified both VZV gH and gL. Transfection experiments showed that rec-RC IgG recognized a VZV gH/gL protein complex but not individual gH or gL proteins. Overall, our recombinant monoclonal anti-VZV antibody effectively neutralizes VZV and recognizes a conformational epitope within the VZV gH/L protein complex. An unlimited supply of this antibody provides the opportunity to analyze membrane fusion events that follow virus attachment and to identify multiple epitopes on VZV-specific proteins.  相似文献   
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Biophysical-Immunological Assay for Ribonucleic Acid Type C Viruses   总被引:3,自引:0,他引:3       下载免费PDF全文
A biophysical and immunological method for characterization of ribonucleic acid type C virus suspensions is described. The method provides a relationship to the total viral mass concentration of the particle titer, the gs antigen titer, and the ultraviolet absorbance (268 nm) of 2% sodium dodecyl sulfate digests. Data for murine, rat, feline, and hamster viruses are shown to be analogous within the test limitations. From these data, an assessment of the viral purity can be made, the structural integrity can be evaluated, an approximate molecular weight can be computed, and the mole ratio of gs antigen can be determined.  相似文献   
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Use of disposable products in hospitals continues to increase despite limited landfill space and dwindling natural resources. We analyzed the use and disposal patterns of disposable hospital products to identify means of reducing noninfectious, nonhazardous hospital waste. In a 385-bed private teaching hospital, the 20 disposable products of which the greatest amounts (by weight) were purchased, were identified, and total hospital waste was tabulated. Samples of trash from three areas were sorted and weighed, and potential waste reductions from recycling and substituting reusable items were calculated. Business paper, trash liners, diapers, custom surgical packs, paper gowns, plastic suction bottles, and egg-crate pads were among the 20 top items and were analyzed individually. Data from sorted trash documented potential waste reductions through recycling and substitution of 78, 41, and 18 tonnes per year (1 tonne = 1,000 kg = 1.1 tons) from administration, the operating room, and adult wards, respectively (total hospital waste was 939 tonnes per year). We offer specific measures to substantially reduce nonhazardous hospital waste through substitution, minimization, and recycling of select disposable products.  相似文献   
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