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  • 1
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    Richmond, Va. : Periodicals Archive Online (PAO)
    Journal of the American Musicological Society. 42:2 (1989:Summer) 432 
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  • 2
    ISSN: 1617-4623
    Keywords: Nitrate reductase ; cDNA expression cloning ; Tobacco ; Sequence ; Cytochrome b5
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Messenger RNAs encoding the nitrate reductase apoenzyme from tobacco can be translated in a cell-free system. Poly(A)+ mRNA fractions from the 23-32 S area of a sucrose gradient were used to build a cDNA library in the expression vector λgt11 with an efficiency of cloning of approximately 104 recombinants/ng mRNA. Recombinant clones were screened with a rabbit polyclonal antibody directed against the corn nitrate reductase, which cross reacts specifically with the nitrate reductases from dicotyledons. Among 240000 recombinant plaques, eight clones were isolated containing inserts of sizes ranging from 1.6 kb to 2.1 kb and sharing sequence homologies. Seven of these clones contained a common internal 1.6 kb EcoRI fragment. The identity of these clones was confirmed as follows. A fusion protein of 170 kDa inducible by IPTG and recognized by the rabbit nitrate reductase antibody was expressed by a lysogen derived from one of the recombinants. The antibodies binding the fused protein were eluted and shown to be inhibitory to the catalytic activity of tobacco nitrate reductase. Two monoclonal antibodies directed against nitrate reductase were also able to bind the hybrid protein. The 1.6 kb EcoRI fragment was sequenced by the method of Sanger. The open reading frame corresponding to a translational fusion with the β-galactosidase coding sequence of the vector shared strong homology at the amino acid level with the heme-binding domain of proteins of the cytochrome b5 superfamily and with human erythrocyte cytochrome b5 reductase. When the 1.6 kb EcoRI fragment was used as a probe for Northern blot experiments a signal corresponding to a 3.5 kb RNA was detected in tobacco and in Nicotiana plumbaginifolia mRNA preparations but no cross-hybridization with corn mRNAs was detected. The probe hybridized with low copy number sequences in genomic blots of tobacco DNA.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 0032-8332
    Keywords: Menstrual cycle ; Vaginal secretion ; Chemical signals ; Pheromones ; Social/physical contact ; Olfactory cues ; Crab-eating monkey ; Cynomolgus monkey ; Breeding husbandry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Twelve adult female crab-eating monkeys (Macaca fascicularis) were placed in six pairs of adjacent cages, allowing physical contact between members of these experimental pairs. Twelve additional females remained singly caged (no physical contact allowed) and served as six control “pairs.” In both experimental and control pairs, one member had a history of regular menstrual cycles, whereas the other tended to have cycles that were unusually long and/or irregular. Over a six-month period, menses and amount of vaginal secretion were recorded daily for all subjects, and the behavior of experimental pairs was sampled three times per week. During the course of the study, the irregular experimental subjects began to exhibit menstrual cycles of near normal length although there was no apparent trend to synchronize cycles. Irregular controls continued to show cycles that were abnormally long. Vaginal secretions tended to increase in all regularly cycling animals during days 9–15 (peak day 11) or reverse days 21–16 of the cycle, consistent with the estimated time of ovulation. Analysis of behavior indicated that irregularly cycling subjects inspected the genitalia of their regularly cycling cagemates at a significantly higher frequency than the converse (F=12.61,p〈.005), particularly during the follicular phase (F=3.39,p〈.07). These results suggest that close physical contact may serve to transmit chemical and/or hormonal cues that can normalize the menstrual cycle of crab-eating monkeys.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1617-4623
    Keywords: Mutagenized protoplast cultures ; Nicotiana plumbaginifolia ; Nitrate-deficient mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two hundred and eleven nitrate reductase-deficient mutants (NR−) were isolated from mutagenized Nicotiana plumbaginifolia protoplast cultures by chlorate selection and regenerated into plant. More than 40% of these clones were classified as cnx and presumed to be affected in the biosynthesis of the molybdenum cofactor, the remaining clones being classified as nia mutants. A genetic analysis of the regenerated plants confirmed this proportion of nia and cnx clones. All mutants regenerated were found to carry monogenic recessive mutations that impaired growth on nitrate as sole nitrogen source. Mutants propagated by grafting on N. tabacum systematically displayed a chlorotic leaf phenotype. This chlorosis was therefore related to the NR deficiency. The observation of leaves with NR− chlorotic sectors surrounded by NR+ wild-type tissues suggeests that an NR deficiency is not corrected by diffusible factors. Periclinal chimeras between wild-type tobacco and the NR− graft were also observed. In this type of chimeric tissue chlorosis was no longer detectable when NR+ cells were in the secondmost (L2) layer, but was still detectable when NR− cells were in the secondmost layer. The genetic analysis of nia mutants revealed that they belong to a single complementation group. However three nia mutants were found to complement some of the other nia mutants. The apoenzyme of nitrate reductase was immunologically detected in several nia mutants but not in other members of this complementation group. Some of the nia mutants, although they were NR−, still displayed methylviologenitrate reductase activity at a high level. These data show that the nia complementation group corresponds to the structural gene of nitrate reductase. Some of the mutations affecting this structural gene result in the overproduction of an inactive nitrate reductase, suggesting a feedback regulation of the level of the apoenzyme in the wild type.
    Type of Medium: Electronic Resource
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