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31.
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
32.
E. F. Woods 《The Biochemical journal》1969,113(1):39-47
Paramyosin from the oyster Crassostrea commercialis was studied by equilibrium sedimentation. In non-denaturing solvents the minimum molecular weight is 208000. Dissociation into subunits requires complete disruption of the alpha-helix. This occurs at pH7 in guanidine hydrochloride solutions of concentration greater than 7m in the presence of a disulphide-bond-reducing agent. Solutions of the protein in concentrated guanidine hydrochloride are polydisperse and contain species of low molecular weight (approx. 25000) comprising approx. 5% to 10% of the protein. The molecular weight of the main component is estimated to be 97000 and the paramyosin molecule contains two of these subunits. From the present observations no decision can be made as to whether or not the small component (or components) represents part of the paramyosin molecule. Preferential binding of guanidine hydrochloride to the extent of 0.13g./g. of protein was shown in solutions of paramyosin in 7.85m-guanidine hydrochloride. 相似文献
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37.
Nucleotide sequence and expression of a cloned Thiobacillus ferrooxidans recA gene in Escherichia coli 总被引:8,自引:0,他引:8
The nucleotide sequence of the recA gene of Thiobacillus ferrooxidans has been determined. No SOS box characteristic of LexA-regulated promoters could be identified in the 196-bp region upstream from the coding region. The cloned T. ferrooxidans recA gene was expressed in Escherichia coli from both the lambda pR and lac promoters. It was not expressed from the 2.2-kb of T. ferrooxidans DNA preceding the gene. The T. ferrooxidans recA gene specifies a protein of 346 amino acids that has 66% and 69% homology to the RecA proteins of E. coli and Pseudomonas aeruginosa, respectively. Most amino acids that have been identified as being of functional importance in the E. coli RecA protein are conserved in the T. ferrooxidans RecA protein. Although some amino acids that have been associated with proteolytic activity have been substituted, the cloned protein has retained protease activity towards the lambda and E. coli LexA repressors. 相似文献
38.
Nucleotide sequence of the Vibrio alginolyticus calcium-dependent, detergent-resistant alkaline serine exoprotease A 总被引:1,自引:0,他引:1
The nucleotide sequence of the Vibrio alginolyticus alkaline serine exoprotease A (ProA) gene cloned in Escherichia coli was determined. The exoprotease A gene (proA) consisted of 1602 bp which encoded a protein of 534 amino acids (aa) with an Mr of 55,900. The region upstream from the gene was characterized by a putative promoter consensus region (-10 -35), a ribosome-binding site and ATG start codon. The proA gene encodes a typical 21-aa N-terminal signal sequence which, when fused to alkaline phosphatase by means of transposon TnphoA, was able to mediate transport of the alkaline phosphatase to the periplasm in E. coli. Deletions of up to 106 aa from the C terminus of ProA did not result in the loss of extracellular protease activity. Additional V. alginolyticus genes were not involved in the secretion into the medium of the cloned ProA in E. coli. The amino acid sequence of ProA showed low overall homology to a Serratia marcescens serine exoprotease but significant homology was detected with other subtilisin family exoproteases. The fungal proteinase K, another sodium dodecyl sulfate-resistant protease, had 44% aa homology with ProA. 相似文献
39.
E W Johnson S W Woods S Zoghbi B J McBride R M Baldwin R B Innis 《Life sciences》1990,47(17):1535-1546
The binding of an iodinated benzodiazepine (BZ) radioligand has been characterized, particularly in regard to its potential use as a neuroreceptor brain imaging agent with SPECT (Single Photon Emission Computed Tomography). Ro16-0154 is an iodine-containing BZ antagonist and a close analog of Ro15-1788. In tissue homogenates prepared from human and monkey brain, the binding of 125I-labeled Ro16-0154 was saturable, of high affinity (Kd = 0.5 nM at 37 degrees C), and had high ratios of specific to non-specific binding (approximately 40:1). Physiological concentrations of NaCl (150 mM) enhanced specific binding approximately 15% compared to buffer without this salt. Kinetic studies of association and dissociation demonstrated a temperature dependent decrease in affinity with increasing temperature. Drug displacement studies confirmed that 125I-Ro16-0154 binds to the "central" type BZ receptor: binding is virtually identical to that of 3H-Ro15-1788 except that 125I-Ro16-0154 shows an almost 10 fold higher affinity at 37 degrees C. These in vitro results suggest that 123I-labeled Ro16-0154 shows promise as a selective, high affinity SPECT probe of the brain's BZ receptor. 相似文献
40.
Serum samples collected from feral and domestic swine (Sus scrofa) in Florida and feral swine in Georgia and Texas were assayed by plaque reduction for their virus neutralizing (VN) antibodies against the porcine transmissible gastroenteritis virus (TGE). None of 560 samples collected from feral swine contained VN antibodies for TGE virus, but experimentally infected feral swine seroconverted. None of 665 samples from domestic swine contained TGE-VN antibodies. These results indicate feral swine are not a significant reservoir for TGE virus in southern states, but are capable of becoming infected and developing VN antibodies against TGE. 相似文献