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Human noroviruses (HuNoV) are a major cause of nonbacterial gastroenteritis worldwide, yet details of the life cycle and replication of HuNoV are relatively unknown due to the lack of an efficient cell culture system. Studies with murine norovirus (MNV), which can be propagated in permissive cells, have begun to probe different aspects of the norovirus life cycle; however, our understanding of the specific functions of the viral proteins lags far behind that of other RNA viruses. Genome-wide functional profiling by insertional mutagenesis can reveal protein domains essential for replication and can lead to generation of tagged viruses, which has not yet been achieved for noroviruses. Here, transposon-mediated insertional mutagenesis was used to create 5 libraries of mutagenized MNV infectious clones, each containing a 15-nucleotide sequence randomly inserted within a defined region of the genome. Infectious virus was recovered from each library and was subsequently passaged in cell culture to determine the effect of each insertion by insertion-specific fluorescent PCR profiling. Genome-wide profiling of over 2,000 insertions revealed essential protein domains and confirmed known functional motifs. As validation, several insertion sites were introduced into a wild-type clone, successfully allowing the recovery of infectious virus. Screening of a number of reporter proteins and epitope tags led to the generation of the first infectious epitope-tagged noroviruses carrying the FLAG epitope tag in either NS4 or VP2. Subsequent work confirmed that epitope-tagged fully infectious noroviruses may be of use in the dissection of the molecular interactions that occur within the viral replication complex. 相似文献
5.
Sign language studies of cross-fostered chimpanzees measure the effect of special rearing conditions on the development of
very young chimpanzees. Cross-fostered chimpanzees, like human children, develop gradually in a process that takes many years.
Here we discuss details of the procedure, the overlap between human and chimpanzee infants in the contents of the first 50-item
vocabularies, and the ways in which the signs of the chimpanzees exhibit the fuzziness of natural language categories. We
also compare the cross-fostering approach with more traditional modular approaches to the study of language-like behavior
in nonhuman animals.
Project Washoe was originally supported by grants MH-12154 from the National Institute of Mental Health and GB-7432 from the
National Science Foundation. We gratefully acknowledge this support and the support that later sign language studies of chimpanzees
have received since then from NIH, NSF, the National Geographic Society, the Grant Foundation, the Spencer Foundation, the
University of Nevada, and the UNR Foundation. 相似文献
6.
A method for identifying cysteine-containing peptides in proteins is presented using 2-bromoacetamido-4-nitrophenol (BNP) to introduce an easily detectable probe. The formation of a covalent bond between the protein sulfhydryl group and the acetamido moiety of BNP introduces a chromophore with an absorbance maximum at 410 nm. The modified protein can then be cleaved with appropriate proteases and the resulting peptides separated by chromatographic methods. Monitoring the effluent at a single wavelength (405 nm) provides a rapid and simple method of detecting and isolating only those peptides which contain cysteine residue(s). The nitrophenol derivative is stable under conditions required for protease cleavage. The reagent is therefore useful for locating cysteine-containing peptides in protein digests and can be used to explore the accessibility of different cysteines under a variety of conditions. The ease of modification, specificity of reaction, product stability, and simple detection of modified peptides make BNP ideal for investigation of cysteine residues. 相似文献
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Genes for serum amyloid A proteins map to Chromosome 7 in the mouse 总被引:10,自引:0,他引:10
Summary Several restriction fragment length variants have been detected among inbred strains using a mouse serum amyloid A cDNA clone. Five variants were shown to segregate as a single genetic unit and were mapped to Chromosome 7 between the glucose phosphate isomerase locus (Gpi-1) and the pink eye dilution locus (p) using recombinant inbred and congenic strains. The finding that no major MspI or BclI restriction fragments were shared between digests of DNAs from a Chromosome 7 congenic strain and its inbred partner, indicate that most, and probably all, sequences detected with the probe are clustered on Chromosome 7. Aneuploid mapping was used to show that the serum amyloid A gene complex (Saa) is proximal to the Chromosome 7 breakpoint in T(7;X)1Ct, a translocation in which the middle third of Chromosome 7 is inserted into the X-chromosome. A survey of inbred strains revealed a single common Saa haplotype and eight rare haplotypes. The complex distribution of 14 different variants suggests that recombination may have played a role in haplotype evolution.This work was supported by grants GM18684 and CA33093 from the National Institute of General Medical Sciences and the National Cancer Institute, respectively. 相似文献
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A neonate with aniridia was found to have a one band paracentric inversion of the short arm of chromosome 4. This was initially difficult to interpret on high resolution banding. The inversion was present in three generations of the family. 相似文献
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Plants often suffer reductions in fecundity due to insect herbivory. Whether this loss of seeds has population-level consequences
is much debated and often unknown. For many plants, particularly those with long-lived seedbanks, it is frequently asserted
that herbivores have minimal impacts on plant abundance because safe-site availability rather than absolute seed number determines
the magnitude of future plant recruitment and hence population abundance. However, empirical tests of this assertion are generally
lacking and the interplay between herbivory, spatio-temporal variability in seed- or safe-site-limited recruitment, and seedbank
dynamics is likely to be complex. Here we use a stochastic simulation model to explore how changes in the spatial and temporal
frequency of seed-limited recruitment, the strength of density-dependent seedling survival, and longevity of seeds in the
soil influence the population response to herbivory. Model output reveals several surprising results. First, given a seedbank,
herbivores can have substantial effects on mean population abundance even if recruitment is primarily safe-site-limited in
either time or space. Second, increasing seedbank longevity increases the population effects of herbivory, because annual
reductions in seed input due to herbivory are accumulated in the seedbank. Third, population impacts of herbivory are robust
even in the face of moderately strong density-dependent seedling mortality. These results imply that the conditions under
which herbivores influence plant population dynamics may be more widespread than heretofore expected. Experiments are now
needed to test these predictions.
Received: 3 November 1999 / Accepted: 15 February 2000 相似文献