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  • 1
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; DNA restriction and cloning ; Gene fusions ; Leucine biosynthesis ; Mutant complementation ; Nostoc ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genomic DNA extracted from the symbiotically-competent, heterocyst-forming cyanobacterium Nostoc sp. strain 7801 was resistant to cleavage by a number of restriction endonucleases. A cosmid library of Nostoc DNA was prepared and maintained in the modification-limited Escherichia coli strain HB101. Analysis of cloned Nostoc DNA fragments indicated infrequent occurrence of restriction endonuclease recognition sites in the Nostoc genome. The Nostoc genomic library was screened for sequences complementing mutations in the E. coli leucine and proline biosynthetic operons. Two cosmids complementing leuB were isolated but none for leuA, leuC, leuD, or proA were detected in 1000 cosmids. A 3.0 kb fragment subcloned from one of the cosmids complemented mutations in leuB when inserted into the HindIII site of pBR322 in either orientation, demonstrating that transcription of leuB originated within the cloned fragment. The cloned fragment also carries a second site capable of initiating transcription of fused antibiotic resistance genes. While transcription of Nostoc DNA sequences did occur in E. coli, unknown barriers must also exist that prevented additional biological complementation of specific E. coli mutations.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of cancer research and clinical oncology 110 (1985), S. 115-118 
    ISSN: 1432-1335
    Keywords: 6-Thioguanine ; 6-Thioguanosine ; 6-Thioguanine riboside ; 6-Mercaptopurine ; Thiopurines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary 6-Thioguanine (6TG) is poorly absorbed after oral administration. Bolus injections of 6TG result in high peak concentrations with relatively shortlived plasma concentrations. In vitro studies have shown the importance of prolonged exposure to 6TG. Therefore we administered 6TG by infusion at a dose rate of 2 mg/h over 2 h. In three goats we determined the plasma concentration-time curves of 6TG and its riboside (6TGR). A steady state was reached for 6TG and was almost reached for 6TGR within the 2 h of infusion. In one experiment we obtained several samples of CSF and observed good penetration of 6TG and 6TGR into CSF. Urinary excretion of 6TG and 6TGR was also quantitated. The amount of drug and metabolite excreted later than 4 h after the end of the infusion was negligible. By infusing 6TG, the problems of both erratic absorption after oral administration and acute renal toxicity after bolus injection, can be averted. In our opinion prolonged infusions of 6TG may be of advantage in humans suffering from actively proliferating malignant diseases, and thus should be studied.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1617-4623
    Keywords: Cyanobacteria ; Nitrate reductase mutants ; Nitrite reductase mutants ; Regulatory mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Eighteen mutant strains of the unicellular cyanobacterium Anacystis nidulans R2 that are unable to assimilate nitrate have been isolated after transposon Tn901 mutagenesis. Characterization of phenotypes and transformation tests have allowed the distinction of five different mutant types. The mutants exhibiting a nitrate reductase-less phenotype were identified as being affected in previously defined loci, as they could be transformed to the wild type by one of the plasmids pNR12, pNR63 or pNR193, which contain cloned genes of A. nidulans R2 involved in nitrate reduction. The mutations in strains FM2 and FM16 appear to affect two other genes involved in nitrate assimilation. Strain FM2 apparently bears a single mutation which results in both lack of nitrite reductase activity and loss of ammonium-promoted repression of nitrate reductase synthesis. FM16 has a low but significant level of nitrate reductase that is also freed from repression by ammonium, and an increased level of nitrite reductase activity. FM16 exhibited properties which indicate that this mutant strain might also be affected in the transport of nitrate into the cell.
    Type of Medium: Electronic Resource
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