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Studies on mutants affecting amidophosphoribosyltransferase activity in Saccharomyces cerevisiae 总被引:1,自引:0,他引:1
Mutants at the ade4 locus of yeast were isolated following mutagenesis of ade+ and ade2 with ultraviolet light (UV), ethylmethane sulphonate, and the acridine half mustard ICR-170. Tests for interallelic complementation, osmotic remediality, temperature sensitivity, and mutagen-specific reversion were carried out on 19 mutants. Six mutants showed interallelic complementation and fell into four groups, defining three complons. Three mutants were osmotic remedial and the same three were temperature sensitive. Three mutants induced by ICR-170 gave purine-excreting revertants, designated Pur6 or ade4.RCF, after exposure to UV. Activity of amidophosphoribosyltransferase (PRPPAT) was assayed in the ade4 mutants and other alleles at this locus. The ade4 mutants lacked activity of the enzyme; the alleles su-pur+, su-pur, PUR6, and Pur6, showed different levels of activity. The enzyme was subject to feedback inhibition by AMP and IMP in su-pur+ and PUR6; su-pur was hypersensitive to inhibition by AMP, whereas Pur6 was slightly resistant. Purine synthesis de novo was shown to be repressible in su-pur+ and constitutive in PUR6 and Pur6 by following the accumulation of aminoimidazole ribotide in the presence and absence of cycloheximide. These observations were confirmed by direct assay of enzyme activity. 相似文献
45.
I D Craig D T Shum P Desrosiers C McLeod M S Lefcoe N A Paterson R J Finley B Woods R J Anderson 《Acta cytologica》1983,27(6):647-650
A case of choriocarcinoma metastatic to the lung following a previous hydatidiform mole is presented. It was possible to make definitive identification of trophoblastic elements on a needle aspiration biopsy using an immunoperoxidase staining technique, thus avoiding diagnostic thoracotomy prior to therapeutic intervention. A method of immunoperoxidase staining of previously fixed and Papanicolaou-stained needle aspiration biopsy specimens is also described, and other uses of the immunoperoxidase technique on needle biopsy specimens are discussed. 相似文献
46.
Transformation of Clostridium acetobutylicum Protoplasts with Bacteriophage DNA 总被引:6,自引:3,他引:3
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Sharon J. Reid Errol R. Allcock David T. Jones David R. Woods 《Applied microbiology》1983,45(1):305-307
Techniques for the transformation of Clostridium acetobutylicum protoplasts with bacteriophage DNA are described. Transformation required regeneration of protoplasts and a 2-h eclipse period. 相似文献
47.
Robin A. Woods Darlene G. Roberts Theodore Friedman Douglas Jolly David Filpula 《Molecular & general genetics : MGG》1983,191(3):407-412
Summary Yeast mutants lacking activity of the enzyme hypoxanthine: guanine phosphoribosyltransferase (H:GPRT) have been isolated by selecting for resistance to 8-azaguanine in a strain carrying the wild type allele, ade4
+ of the gene coding for amidophosphoribosyltransferase (PRPPAT), the first enzyme of de novo purine synthesis. The mutants excrete purines and are cross-resistant to 8-azaadenine. They are recessive and represent a single complementation group, designated hpt1. Ade4-su, a prototrophic allele of ade4 with reduced activity of PRPPAT, is epistatic to hpt1, suppressing purine excretion and resistance to azaadenine but not resistance to azaguanine. The genotype ade2 hpt1 does not respond to hypoxanthine. Hpt1 complements and is not closely linked to the purine excreting mutants pur1 to pur5. Hpt1 and pur6, a regulatory mutant of PRPPAT, are also unlinked but do not complement, suggesting a protein-protein interaction between H:G-PRT and PRPPAT. Mycophenolic acid (MPA), an inhibitor of de novo guanine nucleotide synthesis, inhibits the growth of hpt1 and hpt1
+. Xanthine allows both genotypes to grow in the presence of MPA whereas guanine only allows growth of hpt1
+. Activity of A-PRT, X-PRT and H:G-PRT is present in hpt
+. Hpt1 lacks activity of H:G-PRT but has normal A-PRT and X-PRT. 相似文献
48.
The site of a hypothesized embryo reduction mechanism was studied. The number of normal-sized embryos and undersized embryos (>2 standard deviations below the mean embryo diameter in control mares) was determined at day 7 and 11 post-ovulation in single-ovulating control mares and in multiple-ovulating pituitary extract-treated mares in which all ovulations occurred on a single day. In two additional groups (control and treated), the embryonal enlargement was monitored by rectal palpation until day 49. An average of 0.7 normal-sized embryos per control mare at day 7 and at day 11 and 0.6 vesicles per control mare at day 49 was found. More normal-sized embryos (P<0.05) were recovered per extract-treated mare at day 7 (2.9 embryos/mare) than at day 11 (0.7), and fewer undersized embryos per extract-treated mare (P<0.05) were recovered at day 7 (0.1) than at day 11 (1.1). The total number of embryos (normal-sized plus undersized) per treated mare was greater (P<0.05) at day 7 and at day 11 than the number of palpated vesicles per treated mare at day 49. The number of mares with more than one normal-sized embryo in the day 7 treated group (10/14) was greater (P<0.05) than in the day 11 treated group (1/14) and was greater than the number of mares with more than one palpated vesicle in the day 49 treated group (2/14). Intrauterine reduction was therefore manifested between days 7 and 11 in multiple-ovulating mares, as demonstrated by the number of multiple normal-sized blastocysts recovered at day 7 and by the reduced number of normal-sized and the increased number of undersized blastocysts recovered at day 11. 相似文献
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Robin Arthur Woods 《Molecular & general genetics : MGG》1969,105(4):314-316
Summary Confirmation that the ad-2 locus of yeast controls the carboxylation of aminoimidazole ribonucleotide (AIR) to 5-amino-4-imidazole carboxylate ribonucleotide (CAIR) is provided by the observation that 21 out of a sample of 113 ad-2 mutants were affected by CO2. 19 of the mutants were stimulated by CO2 and 2 were inhibited. The majority of the CO2-stimulated mutants were confined to one section of the complementation map of the ad-2 locus. 相似文献