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  • American Society for Microbiology  (11)
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  • American Society for Microbiology  (11)
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  • 1
    Online Resource
    Online Resource
    Characterization of proviruses cloned from mink cell focus-forming virus-infected cellular DNA
    American Society for Microbiology ; 1982
    In:  Journal of Virology Vol. 41, No. 2 ( 1982-02), p. 435-448
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 41, No. 2 ( 1982-02), p. 435-448
    Abstract: Two proviruses were cloned from EcoRI-digested DNA extracted from mink cells chronically infected with AKR mink cell focus-forming (MCF) 247 murine leukemia virus (MuLV), using a lambda phage host vector system. One cloned MuLV DNA fragment (designated MCF 1) contained sequences extending 6.8 kilobases from an EcoRI restriction site in the 5' long terminal repeat (LTR) to an EcoRI site located in the envelope (env) region and was indistinguishable by restriction endonuclease mapping for 5.1 kilobases (except for the EcoRI site in the LTR) from the 5' end of AKR ecotropic proviral DNA. The DNA segment extending from 5.1 to 6.8 kilobases contained several restriction sites that were not present in the AKR ecotropic provirus. A 0.5-kilobase DNA segment located at the 3' end of MCF 1 DNA contained sequences which hybridized to a xenotropic env-specific DNA probe but not to labeled ecotropic env-specific DNA. This dual character of MCF 1 proviral DNA was also confirmed by analyzing heteroduplex molecules by electron microscopy. The second cloned proviral DNA (designated MCF 2) was a 6.9-kilobase EcoRI DNA fragment which contained LTR sequences at each end and a 2.0-kilobase deletion encompassing most of the env region. The MCF 2 proviral DNA proved to be a useful reagent for detecting LTRs electron microscopically due to the presence of nonoverlapping, terminally located LTR sequences which effected its circularization with DNAs containing homologous LTR sequences. Nucleotide sequence analysis demonstrated the presence of a 104-base-pair direct repeat in the LTR of MCF 2 DNA. In contrast, only a single copy of the reiterated component of the direct repeat was present in MCF 1 DNA.
    Type of Medium: Online Resource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.41.2.435-448.1982
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 1982
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  • 2
    Online Resource
    Online Resource
    Effects of Environmental pH on Membrane Proteins in Borrelia burgdorferi
    American Society for Microbiology ; 1999
    In:  Infection and Immunity Vol. 67, No. 7 ( 1999-07), p. 3181-3187
    Details
    In: Infection and Immunity, American Society for Microbiology, Vol. 67, No. 7 ( 1999-07), p. 3181-3187
    Abstract: Borrelia burgdorferi , the causative agent of Lyme disease, alternates between the microenvironments of the tick vector, Ixodes scapularis , and a mammalian host. The environmental conditions the spirochete encounters during its infectious cycle are suspected to differ greatly in many aspects, including available nutrients, temperature, and pH. Here we identify alterations in the membrane protein profile, as determined by immunoblotting and two-dimensional nonequilibrium pH gradient gel electrophoresis (2D-NEPHGE), that occur in virulent B. burgdorferi B31 as the pH of the medium is altered. Initial comparisons of cultures incubated at pHs 6.0, 7.0, and 8.0 yielded alterations in the expression of seven membrane proteins as determined by probing with hyperimmune rabbit serum. Six of these membrane proteins (54, 45, 44, 43, 35, and 24 kDa) were either present in increased amounts in or solely expressed by cultures incubated at pHs 6.0 and 7.0. The 24-kDa protein that decreased in expression at pH 8.0 was identified as outer surface protein C (OspC). In addition, a 42-kDa membrane protein increased in amount in cultures incubated at pH 8.0. Similar changes were observed with serum from a mouse infected by tick bite, with the recognition of two additional bands (48 and 46 kDa) unique to pHs 6.0 and 7.0. When membrane fractions were analyzed by 2D-NEPHGE, at least 37 changes in the membrane protein profile between cells incubated at pHs 6.0, 7.0, and 8.0 were observed by immunoblotting and silver staining. Environmental cues such as pH may prove important in the regulation of virulence determinants and factors necessary for the adaptation of B. burgdorferi to the tick or mammalian microcosm.
    Type of Medium: Online Resource
    ISSN: 0019-9567 , 1098-5522
    URL: Article
    DOI: 10.1128/IAI.67.7.3181-3187.1999
    RVK:
    XA 10000
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 1999
    detail.hit.zdb_id: 1483247-1
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  • 3
    Online Resource
    Online Resource
    Molecular cloning of the Harvey sarcoma virus closed circular DNA intermediates: initial structural and biological characterization
    American Society for Microbiology ; 1979
    In:  Journal of Virology Vol. 31, No. 3 ( 1979-09), p. 795-809
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 31, No. 3 ( 1979-09), p. 795-809
    Abstract: Supercoiled Harvey sarcoma virus (Ha-SV) DNA was extracted from newly infected cells by the Hirt procedure, enriched by preparative agarose gel electrophoresis, and digested with EcoRI, which cleaved the viral DNA at a unique site. The linearized Ha-SV DNA was then inserted into lambda gtWESlambda B at the EcoRI site and cloned in an approved EK2 host. Ha-SV DNA inserts from six independently derived recombinant clones have been analyzed by restriction endonuclease digestion, molecular hybridization, electron microscopy, and infectivity. Four of the Ha-SV DNA inserts were identical, contained about 6.0 kilobase pairs (kbp), and comigrated in agarose gels with the infectious, unintegrated, linear Ha-SV DNA. One insert was approximately 0.65 kbp smaller (5.35 kbp) and one was approximately 0.65 kpb larger (6.65 kpb) than the 6.0 kpb inserts. R-looping with Ha-SV RNA revealed that the small (5.35 kbp) insert contained one copy of the Ha-SV RNA. Preliminary restriction endonuclease digestion of the recombinant DNAs suggested that the middle-size inserts contained a 0.65-kbp tandem duplication of sequences present only one in the small-size insert; this duplication corresponded to the 0.65-kpb terminal duplication of the unintegrated linear Ha-SV DNA. The large-size insert apparently contained a tandem triplication of these terminally located sequences. DNA of all three sized inserts induced foci in NIH 3T3 cells, and focus-forming activity could be rescued from the transformed cells by superinfection with helper virus. Infectivity followed single-hit kinetics, suggesting that the foci were induced by a single molecule.
    Type of Medium: Online Resource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.31.3.795-809.1979
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 1979
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  • 4
    Online Resource
    Online Resource
    Microbiology of Brazilian purpuric fever and diagnostic tests. Brazilian Purpuric Fever Study Group
    American Society for Microbiology ; 1989
    In:  Journal of Clinical Microbiology Vol. 27, No. 4 ( 1989-04), p. 605-608
    Details
    In: Journal of Clinical Microbiology, American Society for Microbiology, Vol. 27, No. 4 ( 1989-04), p. 605-608
    Type of Medium: Online Resource
    ISSN: 0095-1137 , 1098-660X
    URL: Article
    DOI: 10.1128/jcm.27.4.605-608.1989
    RVK:
    XA 10000
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 1989
    detail.hit.zdb_id: 1498353-9
    SSG: 12
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  • 5
    Online Resource
    Online Resource
    Molecular cloning of the Harvey sarcoma virus circular DNA intermediates. II. Further structural analyses
    American Society for Microbiology ; 1980
    In:  Journal of Virology Vol. 33, No. 2 ( 1980-02), p. 845-855
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 33, No. 2 ( 1980-02), p. 845-855
    Abstract: Three species of unintegrated supercoiled Harvey sarcoma virus DNA (6.6, 6.0, and 5.4 kilobase pairs) have been molecularly cloned from Harvey sarcoma virus-infected cells. On the basis of restriction enzyme analyses, the 6.6- and 6.0-kilobase pair viral DNAs contain two and one copies, respectively, of a 650-base pair DNA segment which contains sequences present at the 3' and 5' termini of the viral genome. R-loop structures formed between Moloney leukemia virus RNA and the cloned Harvey sarcoma virus DNA indicated that about 500 base pairs of the 650-base pair repeating segment was complementary to the 3' end of the viral RNA. During amplification in the Escherichia coli host, some recombinants containing the 6.6- or the 6.0-kilobase pair Harvey sarcoma virus DNA insert acquired or lost the complete 650-base pair DNA segment. These changes occurred in both recA+ and recA- E. coli.
    Type of Medium: Online Resource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.33.2.845-855.1980
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 1980
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  • 6
    Online Resource
    Online Resource
    McDonough feline sarcoma virus: characterization of the molecularly cloned provirus and its feline oncogene (v-fms)
    American Society for Microbiology ; 1982
    In:  Journal of Virology Vol. 41, No. 2 ( 1982-02), p. 489-500
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 41, No. 2 ( 1982-02), p. 489-500
    Abstract: The genetic structure of the McDonough strain of feline sarcoma virus (SM-FeSV) was deduced by analysis of molecularly cloned, transforming proviral DNA. The 8.2-kilobase pair SM-FeSV provirus is longer than those of other feline sarcoma viruses and contains a transforming gene (v-fms) flanked by sequences derived from feline leukemia virus. The order of genes with respect to viral RNA is 5'-gag-fms-env-3', in which the entire feline leukemia virus env gene and an almost complete gag sequence are represented. Transfection of NIH/3T3 cells with cloned SM-FeSV proviral DNA induced foci of morphologically transformed cells which expressed SM-FeSV gene products and contained rescuable sarcoma viral genomes. Cells transformed by viral infection or after transfection with cloned proviral DNA expressed the polyprotein (P170gag-fms) characteristic of the SM-FeSV strain. Two proteolytic cleavage products (P120fms and pp55gag) were also found in immunoprecipitates from metabolically labeled, transformed cells. An additional polypeptide, detected at comparatively low levels in SM-FeSV transformants, was indistinguishable in size and antigenicity from the envelope precursor (gPr85env) of feline leukemia virus. The complexity of the v-fms gene (3.1 +/- 0.3 kilobase pairs) is approximately twofold greater than the viral oncogene sequences (v-fes) of Snyder-Theilen and Gardner-Arnstein FeSV. By heteroduplex, restriction enzyme, and nucleic acid hybridization analyses, v-fms and v-fes sequences showed no detectable homology to one another. Radiolabeled DNA fragments representing portions of the two viral oncogenes hybridized to different EcoRI and HindIII fragments of normal cat cellular DNA. Cellular sequences related to v-fms (designated c-fms) were much more complex than c-fes and were distributed segmentally over more than 40 kilobase pairs in cat DNA. Comparative structural studies of the molecularly cloned proviruses of Synder-Theilen, Gardner-Arnstein, and SM-FeSV showed that a region of the feline-leukemia virus genome derived from the pol-env junction is represented adjacent to v-onc sequences in each FeSV strain and may have provided sequences preferred for recombination with cellular genes.
    Type of Medium: Online Resource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.41.2.489-500.1982
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 1982
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  • 7
    Online Resource
    Online Resource
    Structural analysis of integrated polyomavirus DNA in a polyomavirus-induced hamster tumor cell line
    American Society for Microbiology ; 1983
    In:  Journal of Virology Vol. 48, No. 1 ( 1983-10), p. 40-51
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 48, No. 1 ( 1983-10), p. 40-51
    Abstract: The nucleotide sequence and structural organization of integrated polyomavirus DNA and flanking host chromosomal DNA in a polyomavirus-induced hamster tumor cell line were evaluated. Although the proximal portion of the early transcriptional region was intact, the remainder of the integrated viral sequences was characterized by a series of rearrangements, including deletions, insertions, and a tandem duplication. Short direct repeats of different nucleotide sequences located on each side of the host-viral DNA junctions we examined were observed. Tumor cell chromosomal DNA flanking the integration site contained regions of patchy homology with viral DNA sequences which could have participated in the recombinational events leading to stable integration. The presence of several unusual structural features of tumor cell chromosomal DNA at the site of viral DNA integration, which were reminiscent of papovavirus DNA integrated into transformed cells, suggests that the recombinational mechanisms operative in vivo are accurately reflected by those functioning in vitro.
    Type of Medium: Online Resource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.48.1.40-51.1983
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 1983
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  • 8
    Online Resource
    Online Resource
    Identification of 11 pH-Regulated Genes in Borrelia burgdorferi Localizing to Linear Plasmids
    American Society for Microbiology ; 2000
    In:  Infection and Immunity Vol. 68, No. 12 ( 2000-12), p. 6677-6684
    Details
    In: Infection and Immunity, American Society for Microbiology, Vol. 68, No. 12 ( 2000-12), p. 6677-6684
    Abstract: When Borrelia burgdorferi is transmitted from the tick vector to the mammalian host, the bacterium experiences alterations in its environment, such as changes in temperature and pH. Previously, we observed numerous alterations in the membrane protein profile when B. burgdorferi B31 was grown at pH 7.0 compared to pH 8.0. Here we identify 11 genes localizing to linear plasmids that are up-regulated at pH 7.0 relative to pH 8.0 in vitro. Seven genes ( bba03, bba24, bba64, bba66, bbe31, bbj41/bbi39 [encoding products that are 99% identical], and bbk01 ) were indirectly identified by proteomic analysis of membrane proteins. Another gene, bba36 , was identified by screening a B. burgdorferi B31 genomic library with cross-adsorbed hyperimmune rabbit serum. Two additional genes, bba65 and bba73 , were identified by Northern blot analysis. Genes bba64, bba65, bba66, bbj41/bbi39 , and bba73 are members of paralogous gene family 54, and bbe31 is a member of the closely related paralogous gene family 60. Gene bba24 is part of a bicistronic operon with bba25 that encodes the well-characterized decorin binding proteins A and B. All 11 genes were transcriptionally regulated, yet the degree of pH regulation varied, with some genes more tightly regulated than others. The regions upstream of these pH-regulated genes appeared to be unrelated, yet many contained dyad repeats ranging from 12 to 25 nucleotides in length that may be involved in the regulation of these genes.
    Type of Medium: Online Resource
    ISSN: 0019-9567 , 1098-5522
    URL: Article
    DOI: 10.1128/IAI.68.12.6677-6684.2000
    RVK:
    XA 10000
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2000
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  • 9
    Online Resource
    Online Resource
    In vitro synthesis of double-stranded DNA from the Kilham rat virus single-stranded DNA genome
    American Society for Microbiology ; 1978
    In:  Journal of Virology Vol. 27, No. 3 ( 1978-09), p. 784-790
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 27, No. 3 ( 1978-09), p. 784-790
    Abstract: Double-stranded, full-length linear DNA was synthesized in vitro by using single-stranded linear DNA as a self-priming template from the parvovirus Kilham rat virus and Escherichia coli DNA polymerase "large fragment" as the polymerizing enzyme. To ascertain the order of the synthesis of the cleavage fragments and to assess the accuracy of the in vitro synthesis, restriction endonuclease cleavage sites with known recognition sequences were mapped on the DNA. Comparing the cleavage pattern of the synthesized DNA with that of double-stranded viral DNA isolated from infected cells confirms that the in vitro synthesis produces a faithful copy of the viral single-stranded genome. Electron micrographs of the in vitro product reveal it to be a double-stranded linear molecule.
    Type of Medium: Online Resource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.27.3.784-790.1978
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 1978
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  • 10
    Online Resource
    Online Resource
    A cloned polyoma DNA fragment representing the 5' half of the early gene region is oncogenic
    American Society for Microbiology ; 1980
    In:  Journal of Virology Vol. 36, No. 2 ( 1980-11), p. 566-574
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 36, No. 2 ( 1980-11), p. 566-574
    Abstract: The two polyoma DNA fragments generated by cleavage with BamHI and EcoRI were cloned in pBR322, and their oncogenic potential was tested in vivo and in vitro. Only recombinant plasmid DNA containing a polyoma DNA fragment which extends clockwise from 58 to 0 map units and include approximately the 5'-proximal half of the early gene region produced tumors in newborn hamsters and transformed rat embryo cells in tissue culture. Southern blotting analysis indicated that the entire 2.2-kilobase polyoma BamHI-EcoRI fragment was intact in both a tumor cell line and a cell line transformed in culture which we examined. The presence of polyoma middle and small T antigen in these lines was demonstrated by immunoprecipitation and tryptic peptide mapping. DNA from a recombinant plasmid containing a polyoma genome deleted between 90 and 4 map units failed to induce tumors or transform cells.
    Type of Medium: Online Resource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.36.2.566-574.1980
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 1980
    detail.hit.zdb_id: 1495529-5
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