bluuebunny/crossref_metadata_2025 huggingface dataset:
Methyl galactosidase activity: an alternative evolutionary destination for the ebgA0 gene
Authors: Hall B G
Published: January 2020

Previous studies (Campbell et al., 1973; Hall and Hartl, 1974; Hall and Hartl, 1975) have shown that the ebgA0 gene, whose product does not hydrolyze lactose may evolve so that its product does hydrolyze lactose; i.e., lactase activity is one evolutionary destination of the ebgA0 gene. Beginning with a strain that synthesizes ebgA0 gene product constitutively and grows extremely slowly (doubling time, 30 to 50 h) on methyl-beta-D-galactopyranoside (MG), a derivative was selected capable of growth on MG at a moderate rate (doubling time, 5.9 h). Genetic evidence is presented showing that the gene that permits growth on MG is an allele of ebgA. A comparison among strains bearing several alleles of ebgA shows that the new allele, termed ebgAmg, synthesizes a product specific for MG and thus represents a true alternative evolutionary destination for the ebgA0 gene.
Evidence that the isomerization of D-ribose and L-rhamnose is catalyzed by the same enzyme in Mycobacterium smegmatis
Authors: Izumori K , Mitchell M , Elbein A D
Published: January 2020

D-Ribose isomerase was purified and crystallized from cells of Mycobacterium smegmatis grown on either D-ribose or L-rhamnose. Isomerase activity for both of these sugars remained together throughout the purification. The isomerase from L-rhamnose-grown cells had the same chemical and physical properties as the enzyme isolated from D-ribose grown cells. In addition, immunological studies indicated that both activities were in the same protein since antisera prepared against either of the crystals cross-reacted with the other and gave lines of symmetry by the agar gel diffusion method.
Nonribosomal Nature of Novel, Stable Ribonucleic Acid Species Accumulated by Blue-Green Bacteria
Authors: Bonen L. , Allen G. V. , Dobson P. R. , Doolittle W. F.
Published: January 2020

The facultatively chemoheterotrophic blue-green bacterium
Aphanocapsa
6714 accumulates two novel, stable ribonucleic acid species when deprived of sources of carbon and energy. At least one of these species is nonribosomal.
Characterization of an Escherichia coli K-12 F-Con-mutant
Authors: Havekes L M , Hoekstra W P
Published: January 2020

An Escherichia coli K-12 F-mutant defective in conjugation was isolated by means of a zygotic induction enrichment procedure. The recipient ability of the mutant was reduced about 50 times owing to a block in one of the first steps of the conjugation process. In the mutant, cell envelope alterations could not be observed. Sensitivity toward detergents, antibiotics, and phages was unaltered. The mutation appeared to be co-transducible with pyrD. The linkage order in the region of the mutation is origin KL 99-con-pyrD-aroA.
Map of genes for carotenoid and bacteriochlorophyll biosynthesis in Rhodopseudomonas capsulata
Authors: Yen H C , Marrs B
Published: January 2020

The recently discovered gene transfer system of Rhodopseudomonas capsulata was used to construct a genetic map of a region concerned with bacteriochlorophyll and carotenoid production. Mutants blocked in the biosynthesis of these compounds were isolated, and each was characterized on the basis of pigments accumulated during growth under low pO2. One-point, two-point, three-point, and ratio test crosses were performed between various mutant strains, and the results were amenable to conventional genetic analyses. A mapping function was found that related cotransfer frequency to map distance. Seven clusters of mutations, five affecting carotenoid and two affecting bacteriochlorophyll biosynthesis, were arranged in one linkage group. Each cluster of mutations is thought to represent a gene. The length of the mapped region is estimated to be less than 1% of the genome. Cotransfer is observed between markers separated by about 5 to 10 genes.
Regulation of Chorismate mutase-prephenate dehydratase and prephenate dehydrogenase from alcaligenes eutrophus
Authors: Friedrich C G , Friedrich B , Schlegel H G
Published: January 2020

Highly purified enzymes from Alcaligenes eutrophus H 16 were used for kinetic studies. Chorismate mutase was feedback inhibited by phenylalanine. In the absence of the inhibitor, the double-reciprocal plot was linear, yielding a Km for chorismate of 0.2 mM. When phenylalanine was present, a pronounced deviation from the Michaelis-Menten hyperbola occurred. The Hill coefficient (n) was 1.7, and Hill plots of velocity versus inhibitor concentrations resulted in a value of n' = 2.3, indicating positive cooperativity. Chorismate mutase was also inhibited by prephenate, which caused downward double-reciprocal plots and a Hill coefficient of n = 0.7, evidence for negative cooperativity. The pH optimum of chorismate mutase ranged from 7.8 to 8.2; its temperature optimum was 47 C. Prephenate dehydratase was competitively inhibited by phenylalanine and activated by tyrosine. Tyrosine stimulated its activity up to 10-fold and decreased the Km for prephenate, which was 0.67 mM without effectors. Tryptophan inhibited the enzyme competitively. Its inhibition constant (Ki = 23 muM) was almost 10-fold higher than that determined for phenylalanine (Ki = 2.6 muM). The pH optimum of prephenate dehydratase was pH 5.7; the temperature optimum was 48 C. Prephenate dehydrogenase was feedback inhibited by tyrosine. Inhibition was competitive with prephenate (Ki = 0.06 mM) and noncompetitive with nicotinamide adenine dinucleotide. The enzyme was further subject to product inhibition by p-hydroxyphenylpyruvate (Ki = 0.13 mM). Its Km for prephenate was 0.045 mM, and that for nicotinamide adenine dinucleotide was 0.14 mM. The pH optimum ranged between 7.0 and 7.6; the temperature optimum was 38 C. It is shown how the sensitive regulation of the entire enzyme system leads to a well-balanced amino acid production.
Arrangement and Regulation of the Nitrogen Fixation Genes in
Klebsiella pneumoniae
Studied by Derepression Kinetics
Authors: Collmer Alan , Lamborg Marvin
Published: January 2020

Events underlying derepression of the nitrogen fixation (
nif
) genes in
Klebsiella pneumoniae
M5A1 were analyzed in vivo by comparing the effects of selective inhibitors of transcription and translation on subsequent nitrogenase activity (rate of acetylene reduction). When batch cultures were induced for derepression, an 87-min lag separated ammonium ion/oxygen removal and the appearance of activity. To prevent eventual activity by adding inhibitors during this period, it was found necessary to add rifampin, ammonium ion, or chloramphenicol more than 50, 20, or 10 min, respectively, before activity appeared in a parallel control. When these inhibitors were added to cultures in which nitrogenase activity had already appeared, further increase in activity was not stopped by rifampin or ammonium ion until 45 or 20 min, respectively, after addition. Chloramphenicol stopped further increase in nitrogenase activity almost immediately. These data indicated a
nif
operon whose transcription/translation consumes 40 min. When the kinetics of β-galactosidase induction were analyzed under similar conditions, it was found that 7 min separated the initiation of transcription and the first completions of translation. Extrapolating from this, we find the
nif
operon to occupy 17 average gene lengths. It is argued that the
nif
structural genes in
K. pneumoniae
are contained on one operon, and further, that the disparity between the kinetics of inhibition by rifampin and those by ammonium ion suggests regulation at a locus other than the operator.
Pyrimidine dimer excision in surviving and nonsurviving cells of ultraviolet-irradiated cultures of Escherichia coli
Authors: Schenley R L , Fisher W D , Swenson P A
Published: January 2020

We compared dimer excision in viable and nonviable cells fractions separated from Escherichia coli B/r cultures exposed to ultraviolet (UV) irradiation. For cells grown on minimal medium with glycerol as a carbon source, both fractions from the irradiated (20 J/m2, 5% survival) culture excised 60 to 70% of the thymine dimers from prelabeled DNA within 120 min. This percentage was, within experimental error, the same as that obtained from unseparated cultures. When isolated viable and nonviable populations were given a second UV exposure (20 J/m2) both types of cells were again able to excise dimers. The UV survival curve for the isolated viable population indicates that these cells are no more sensitive to radiation than exponentially growing cells not previously exposed to UV. The extent of dimer excision after UV irradiation was also the same in viable and nonviable cells separated from cultures grown on a glucose minimal medium in which both populations excised about 85% of the dimers within 120 min. These results show that the extent of removal of pyrimidine dimer from deoxyribonucleic acid is not precisely correlated with survival of repair-competent bacterial cells after exposure to UV light.
Pathway of thiamine pyrophosphate synthesis in Micrococcus denitrificans
Authors: Sanemori H , Egi Y , Kawasaki T
Published: January 2020

The pathway of thiamine pyrophosphate (TPP) biosynthesis, which is formed either from exogeneously added thiamine or from the pyrimidine and thiazole moieties of thiamine, in Micrococcus denitrificans was investigated. The following indirect evidence shows that thiamine pyrophosphokinase (EC 2.7.6.2) catalyzes the synthesis of TPP from thiamine: (i) [35S]thiamine incubated with cells of this microorganism was detected in the form of [35S]thiamine; (ii) thiamine gave a much faster rate of TPP synthesis than thiamine monophosphate (TMP) when determined with the extracts; and (iii) a partially purified preparation of the extracts can use thiamine, but not TMP, as the substrate. The activities of the four enzymes involved in TMP synthesis from pyrimidine and thiazole moieties of thiamine were detected in the extracts of M. denitrificans. The extracts contained a high activity of the phosphatase, probably specific for TMP. After M. denitrificans cells were grown on a minimal medium containing 3 mM adenosine, which causes derepression of de novo thiamine biosynthesis in Escherichia coli, the activities of the four enzymes involved with TMP synthesis, the TMP phosphatase, and the thiamine pyrophosphokinase were enhanced two- to threefold. These results indicate that TPP is synthesized directly from thiamine without forming TMP as an intermediate and that de novo synthesis of TPP from the pyrimidine and thiazole moieties involves the formation of TMP, followed by hydrolysis to thiamine, which is then converted to TPP directly. Thus, the pathway of TPP synthesis from TMP synthesized de novo in M. denitrificans is different from that found in E. coli, in which TMP synthesized de novo is converted directly to TPP without producing thiamine.
Mechanism of autolysis of Neisseria gonorrhoeae
Authors: Hebeler B H , Young F E
Published: January 2020

The major autolysin(s) of Neisseria gonorrhoeae was solubilized from envelopes by extraction with 2% Triton X-100 containing 0.5 M NaCl. Neither Triton X-100 nor NaCl alone could effectively release the autolysin(s). The major autolysin is N-acetylmuramyl-L-alanine amidase (EC 3.5.1.28). The pH optimum for this reaction was broad, ranging from 5.5 to 8.5. Optimal hydrolysis of peptidoglycan occurred in 2% Triton X-100 in 0.1 M KCl. Attempts to purify the autolysin were unsuccessful. A rapid assay for enzyme activity was developed using radioactive cell walls as a substrate ([3H]diaminopimelic acid).
Volatile factor involved in the dimorphism of Mucor racemosus
Authors: Mooney D T , Sypherd P S
Published: January 2020

Both hyphal and yeastlike development of Mucor racemosus and M. rouxii were demonstrated under 100% N2. Under standardized conditions in yeast extract-peptone-glucose medium, the morphology depended on the N2 flow rate and not on the glucose concentration. The effect was related to the rate of flushing of the atmosphere over the culture medium. The results indicate that a volatile compound produced by Mucor is involved in morphogenesis.
Evidence against the involvement of chemotaxis in swarming of Proteus mirabilis
Authors: Williams F D , Anderson D M , Hoffman P S , Schwarzhoff R H , Leonard S
Published: January 2020

Nonswarming and nonchemotactic mutants of Proteus mirabilis were isolated after mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine or ultraviolet light. These mutants were used in experiments to determine if chemotaxis is involved in the swarming of P. mirabilis. Nonchemotactic mutants failed to form chemotactic bands in a semisolid casein hydrolysate medium, yet they swarmed on the same medium containing 1.5% agar. Nonswarming mutants were attracted towards individual amino acids and components of tryptose. In cross-feeding experiments, no evidence was obtained to indicate the production of a diffusable chemical repellent. In studies with the wild-type P. mirabilis, no clear-cut negative chemotaxis was seen even though three different assays were used and numerous chemicals were tested. Additional evidence against the involvement of chemotaxis in swarming comes from finding that dialysis does not interfere with swarming; swarm cells will swarm immediately when transferred to fresh media, and swarm cells will swarm on an agar-water medium supplemented with a surfactant. These data indicate that chemotaxis is not involved in the swarming of P. mirabilis.
Regulation of tyramine oxidase synthesis in Klebsiella aerogenes
Authors: Okamura H , Murooka Y , Harada T
Published: January 2020

Tyramine oxidase in Klebsiella aerogenes is highly specific for tyramine, dopamine, octopamine, and norepinephrine, and its synthesis is induced specifically by these compounds. The enzyme is present in a membrane-bound form. The Km value for tyramine is 9 X 10(-4) M. Tyramine oxidase synthesis was subjected to catabolite repression by glucose in the presence of ammonium salts. Addition of cyclic adenosine 3',5'-monophosphate (cAMP) overcame the catabolite repression. A mutant strain, K711, which can produce a high level of beta-galactosidase in the presence of glucose and ammonium chloride, can also synthesize tyramine oxidase and histidase in the presence of inducer in glucose ammonium medium. Catabolite repression of tyramine oxidase synthesis was relieved when the cells were grown under conditions of nitrogen limitation, whereas beta-galactosidase was strongly repressed under these conditions. A cAMP-requiring mutant, MK54, synthesized tyramine oxidase rapidly when tyramine was used as the sole source of nitrogen in the absence of cAMP. However, a glutamine synthetase-constitutive mutant, MK94, failed to synthesize tyramine oxidase in the presence of glucose and ammonium chloride, although it synthesized histidase rapidly under these conditions. These results suggest that catabolite repression of tyramine oxidase synthesis in K. aerogenes is regulated by the intracellular level of cAMP and an unknown cytoplasmic factor that acts independently of cAMP and is formed under conditions of nitrogen limitation.
Characterization of intracytoplasmic hydrocarbon inclusions from the hydrocarbon-oxidizing Acinetobacter species HO1-N
Authors: Scott C C , Finnerty W R
Published: January 2020

The ultrastructure of Acinetobacter sp. strain HO1-N grown on hydrocarbon and nonhydrocarbon substrates was compared using thin sections and freeze-etching. Hydrocarbon-grown cells were characterized by the presence of intracytoplasmic membrane-bound hexadecane inclusions. This membrane did not exhibit a typical unit membrane structure but appeared as a monolayer. The freeze-etch technique revealed the internal structure of the hexadecane inclusions and provided evidence for the presence of a smooth-surfaced limiting membrane. Freeze-etching also revealed intracytoplasmic membranes in the hexadecane-grown cells. These ultrastructural modifications were not present in nonhydrocarbon-grown cells. The hexadecane inclusions were isolated from Acinetobacter. Negative-staining of the inclusions revealed electron-transparent vesicles approximating the size of the inclusions seen in whole cells. Freeze-etching of the purified inclusions revealed membrane-bound vesicles. The purified inclusions exhibited a relatively high value of lipid phosphorus to protein. The lipid composition and the electrophoretic banding pattern of the inclusions on sodium dodecyl sulfate-polyacrylamide gels were determined and compared with other membrane fractions (outer membrane and cytoplasmic membrane) previously isolated from this organism.
Localization of glycogen synthetase during differentiation of presumptive cell types in Dictyostelium discoideum
Authors: Harris J F , Rutherford C L
Published: January 2020

Ultramicrochemical techniques were utilized to assay glycogen synthetase (EC 2.4.1.11) activity in cell samples of Dictyostelium discoideum as small as 0.01 mug (dry weight) in reaction volumes of 0.1 mul. The activity was assayed by an amplification procedure employing the enzymatic cycling of pyridine nucleotides. These techniques were used to determine the extent of localization of glycogen synthetase in the two cell types during differentiation of D. discoideum. Localization studies in developing spore cells revealed decreasing enzyme activity to the culmination stage. During this phase of development, the enzyme required the presence of soluble glycogen for activity. From culmination to sorocarp stage, enzyme activity increased and was independent of the soluble glycogen. In developing stalk cells, synthetase showed a decreasing gradient of activity. In sorocarps, the cells in the stalk apex showed synthetase activity similar to that of the spores. The cells at the bottom of the stalk had no detectable activity.
Identification of Several Unique, Low-Molecular-Weight Basic Proteins in Dormant Spores of
Clostridium bifermentans
and Their Degradation During Spore Germination
Authors: Setlow Peter , Waites William M.
Published: January 2020

Two acid-soluble, low-molecular-weight basic proteins comprise ∼20% of the protein in dormant spores of
Clostridium bifermentans
. Both of these proteins are rapidly degraded during spore germination.
Time-Lapse Photography of
Bacillus subtilis
L-Forms Replicating in Liquid Medium
Authors: Gilpin Richard W. , Nagy Stephen S.
Published: January 2020

Phase-contrast observations indicate that an L-form of
Bacillus subtilis
divides in liquid medium by a “budding-like” mechanism without participation of “large bodies” or “elementary bodies.”
Purification and properties of two deoxyribonucleases of Pseudomonas aeruginosa
Authors: Miller R V , Clark A J
Published: January 2020

A survey of the major deoxyribonucleases in Pseudomonas aeruginosa strain PAO was undertaken. Two activities predominated in Brij-58 lysates of this organism. These have been purified from contaminating nuclease activities, and some of their properties have been elucidated. The first was a nuclease that degraded heat-denatured deoxyribonucleic acid (DNA) to mono- and dinucleotides. The activity of this enzyme was confined to single-stranded DNA, and 100% of the substrate was hydrolyzed to acid-soluble material. The Mg2+ optimum is low (1 to 3mM), and the molecular weight is 6 X 10(4). The second predominant activity was an adenosine 5'-triphosphate (ATP)-dependent deoxyribonuclease. This enzyme had an absolute dependence on the presence of ATP Mg2+ concentrations of approximately 10 mM. Five moles of ATP was consumed for each mole of phosphodiester bonds cleaved. The acid-soluble products of the reaction consisted of short oligonucleotides from one to six bases in length. Only 50% of the double-stranded DNA was rendered acid soluble in a limit digest. The molecular weight of this enzyme is 3 X 10(5). The observation of these enzymes in P. aeruginosa is consistent with the possibility that recombinational pathways similar to those of Escherichia coli are operating in this organism.
Phospholipid composition and phospholipase A activity of Neisseria gonorrhoeae
Authors: Senff L M , Wegener W S , Brooks G F , Finnerty W R , Makula R A
Published: January 2020

Exponential-phase cells of Neisseria gonorrhaeae 2686 were examined for phospholipid composition and for membrane-associated phospholipase A activity. When cells were harvested by centrifugation, washed, and lyophilized before extraction, approximately 74% of the total phospholipid was phosphatidylethanolamine, 18% was phosphatidylglycerol, 2% was cardiolipin, and 10% was lysophosphatidylethanolamine. However, when cells still suspended in growth medium were extracted, the amount of lysophosphatidylethanolamine decreased to approximately 1% of the phospholipid composition. This suggests that a gonococcal phospholipase A may be activated by conditions encountered during centrifugation and/or lyophilization of cells preceding extraction. Phospholipase A activity associated with cell membranes was assayed by measuring the conversion of tritiated phosphatidylethanolamine to lysophosphatidylethanolamine. Optimal activity was demonstrated in 10% methanol at pH 8.0 to 8.5, in the presence of calcium ions. The activity was both detergent sensitive and thermolabile. Comparisons of gonococcal colony types 1 and 4 showed no significant differences between the two types with respect to either phospholipid content or phospholipase A activity.
Specialized transduction of D-serine deaminase genes: formation of lysogens that yield high lambda-d dsd/lambda ratios and formation of a dimeric lambda-d dsd
Authors: Palchaudhuri S , McFall E , Carothers A M
Published: January 2020

We have obtained two classes of double lysogens that on induction yield higher titers of lambda-d dsd transducing phage than of helper phage. One class was obtained by lysogenization of strain EM6116 (dsddelta attlambdadelta HfrC) with lambda-dsd type 2 (dsdC+ dsdO+ dsdA+, head-tail substitution). In the absence of either a normal attlambda or the homology of a chromosomal dsd region, the transducing phage integrated at other sites, at least one of which, in strain EM6177, is near the origin of HfrC. On induction, strain EM6177 yields a phage burst of 20 to 50 with a lambdadsd:lambda ratio of 10(4):1. The asnychronously high yield of lambda dsd is attributed to an efficiency of excision greater than that of lambda. The other class was obtained by lysogenization of strain EM1407 (dsdA attlambda+) with lambda-dsd type 2 (dsdO6 dsdA, partial deletion of dsdC). The DNA of mature lambda-dsd type 2 is a complete dimer. It lacks nearly all the phage late genes and b2 and carries about five bacterial genes. It could not be packaged as a monomer but is just within the packaging size limit as a dimer. Models for the derivation of these lambda dsd phages and the high-yielding lysogens are presented.
Stringent control of peptidoglycan biosynthesis in Escherichia coli K-12
Authors: Ishiguro E E , Ramey W D
Published: January 2020

[3H]Diaminopimelic acid (Dap) was incorporated exclusively into peptidoglycan by Escherichia coli strains auxotrophic for both lysine and Dap. The rate of [3H]Dap incorporation by stringent (rel+) strains was significantly decreased when cells were deprived of required amino acids. The addition of chloramphenicol to amino acid-starved rel+ cultured stimulated both peptidoglycan and ribonucleic acid synthesis. In contrast, a relaxed (relA) derivative incorporated [3H]Dap at comparable rates in the presence or absence of required amino acids. Physiologically significant concentrations of guanosine 5'-diphosphate 3'-diphosphate (ppGpp) inhibited the in vitro synthesis of both carrier lipid-linked intermediate and peptidoglycan catalyzed by a particulate enzyme system. The degree of inhibition was dependent on the concentration of ppGpp in the reaction mixture. Thus, the results of in vivo and in vitro studies indicate that peptidoglycan synthesis is stringently controlled in E. coli.
Mycoplasma phosphoenolpyruvate-dependent sugar phosphotransferase system: purification and characterization of the phosphocarrier protein
Authors: Ullah A H , Cirillo V P
Published: January 2020

The Mycoplasma phosphoenolpyruvate-dependent sugar phosphotransferase system consists of three components: a membrane-bound enzyme II, a soluble enzyme I, and a soluble phosphocarrier protein, HPr. The HPr has been purified to homogeneity by a combination of ammonium sulfate precipitations, gel filtration and diethylaminoethyl, carboxymethyl Bio-Gel A, and hydroxylapatite column chromatography. The purified protein is relatively heat stable (ca. 50% activity survives 30 min of boiling) and has a molecular weight of ca. 10,000 (determined by sodium dodecyl sulfate-gel electrophoresis and amino acid analysis). It contains a single histidine residue per molecule and can be totally inactivated by photooxidation with Rose Bengal dye. Although the mycoplasma HPr is very similar to that of Escherichia coli, it shows no significant association with antiserum produced against E. coli HPr.
Role of deoxyribonucleic acid polymerase III in the repair of single-strand breaks produced in Escherichia coli deoxyribonucleic acid by gamma radiation
Authors: Hamelin C , Youngs D A , Smith K C
Published: January 2020

Cell survival, deoxyribonucleic acid (DNA) degradation, and the repair of DNA single-strand breaks were measured for Escherichia coli K-12 pol+, polA1, polC1026(ts), and polA1 polC1026(ts) cells after 137Cs gamma irradiation. The results indicate that DNA polymerase III is required for growth medium-dependent (type III) repair in polA+ or polA cells. In pol+ or polC cells, DNA polymerase I performs type II repair efficiently. The relative deficiencies of each of these strains in DNA repair generally correlate with their relative sensitivities to cell killing and with the extent of DNA degradation observed.
Interference with propagation of typing bacteriophages by extrachromosomal elements in Salmonella typhimurium: bacteriophage type 505
Authors: van Embden J D , van Leeuwen W J , Guinée P A
Published: January 2020

Samonella typhimurium bacteriophage type 505 is the most frequently encountered phage type in the Netherlands and its neighboring countries. Phage type 505 was analyzed with regard o the interference with propagation of the typing phages by the prophages and plasmids, present in the type strain S. typhimurium 505...
Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid
Authors: Meyers J A , Sanchez D , Elwell L P , Falkow S
Published: January 2020

Agarose gel electrophoresis may be employed effectively for the detection and preliminary characterization of plasmid deoxyribonucleic acid (DNA) present in clinical isolates and laboratory strains of gram-negative microorganisms. The method is sensitive and does not require radioisotopes or ultracentrifugation. The estimation of plasmid mass from the extent of DNA migration in gels compares favorably with results obtained by electron microscopy of plasmid DNA purified by equilibrium density centrifugation. The method has proved to be a useful tool for survey work and the epidemiological investigation of plasmid dissemination, as well as an important adjunct to the genetic analysis of plasmids.
Binding of plant lectins to mycoplasma cells and membranes
Authors: Kahane I , Tully J G
Published: January 2020

The binding of iodinated wheat germ agglutinin, Ricinus communis agglutinin, and concanavalin A to mycoplasma cells and membranes was examined. All mycoplasmas studied specifically bound concanavalin A or R. communis agglutinin and, to a lesser degree, wheat germ agglutinin. The binding of lectins to whole cells was similar to that recorded for membranes, suggesting that significant binding only occurred on the outer surface of the mycoplasma membrane. Proteolysis of the membrane almost always increased the capacity to bind lectins, which indicates that additional carbohydrate groups on the mycoplasma membrane are masked by a protein layer or protein complexes on the membrane. The observation that carbohydrates are apparently exposed on the surface of mycoplasma membranes should stimulate more concentrated study on the isolation and chemical characterization of these substances since it is quite likely that they are responsible for a variety of reactions between mycoplasmas and host cells.
Light microscopy of basidia, basidiospores, and nuclei in spores and hyphae of Filobasidiella neoformans (Cryptococcus neoformans)
Authors: Erke K H
Published: January 2020

Three hypha-forming strains of Cryptococcus neoformans were induced to form basidia and basidiospores. Light microscopy showed that basidia formed at the ends of terminal hyphal cells and were able to produce from a few to many basidiospores. The morphology of the sexual structures indicated that these strains belonged to the recently described perfect state of C. neoformans, Filobasidiella neoformans. The average dimensions of the basidiospores were 1.9 mum in width by 2.7 mum in length. Giemsa staining revealed that dikaryotic cells were formed in all three strains. Only one strain had both terminal and subterminal dikaryons, indicating functional clamp connections, whereas the two remaining strains had dikaryons restricted to the terminal cells. Basidiospores of two strains were mononucleate, and yeast cell clones derived from single basidiospores of these two strains were able to complete the sexual life cycle, thus indicating their primary homothallic nature.
Size and transforming activity of deoxyribonucleic acid in Diplococcus pneumoniae during thymidine starvation
Authors: Bousque J L , Sicard N
Published: January 2020

The transforming activity and the molecular structure of DNA from cells of Diplococcus pneumoniae during thymidine starvation have been analyzed and the effects of thymidine starvation have been compared with the effects of single-strand breaks produced by deoxyribonucleases in DNA of unstarved cells. The decrease in transforming activity of lysates from starved cells as a function of the size of DNA particles, measured by centrifugation in neutral and alkaline sucrose gradients, does not follow the kinetics observed after enzymatic degradation of DNA of unstarved cells. Moreover, a strain lacking exo- and endonuclease activities is not protected from thymineless death. These results suggest that the basic lethal mechanism of thymidine starvation might have an origin other than the activation of nucleases.
Isolation and characterization of mutations creating high-efficiency transcription initiation signals within the trp operon of Escherichia coli
Authors: McPartland A , Somerville R L
Published: January 2020

Two different mutational events generate promoter-active deoxyribonucleic acid sequences within the trp operon of Escherichia coli, probably through single base-pair changes. The mutations, obtained after ethyl methane sulfonate mutagenesis by selecting for elevated lac gene expression in a trp-lac fusion, are cis dominant and trans recessive with respect to their effects on the synthesis of downstream enzymes. One of the mutants (trpD11), obtained repeatedly under the selective conditions employed, prevents the formation of active phosphoribosyltransferase. The second mutation, trp3B, has no effects on any trp enzyme. By deletion mapping, trpD11 was localized near the operator-distal end of trpD, outside the segment of deoxyribonucleic acid that contains the low-efficiency internal promoter trpP2. Reversion to prototrophy of trpD11 was greatly stimulated by 2-aminopurine and ethyl methane sulfonate. Tests with suppressors indicated that trpD11 is a UAA (ochre) nonsense mutation. Under repression conditions, strains harboring either lesion in a normal trp operon synthesize the tryptophan biosynthetic enzymes in a noncoordinate fashion. The products of the operator-distal structural genes trpC, trpB, and trpA are formed at rates approximately 15-fold higher than those of wild type. The enzymes encoded by operator-proximal genes trpE and trpD are low or not detectable. Under derepression conditions, coordinate expression of the operon was observed.
Protoplast Formation in
Escherichia coli
Authors: Weiss Richard L.
Published: January 2020

A procedure for protoplast formation in
Escherichia coli
is described. Removal of the cell wall was confirmed by examination of cells in thin-section preparations.
Inorganic nitrogen assimilation by the photosynthetic bacterium Rhodopseudomonas capsulata
Authors: Johansson B C , Gest H
Published: January 2020

The photosynthetic bacterium Rhodopseudomonas capsulata lacks glutamate dehydrogenase and normally uses the glutamine synthetase/glutamate synthase sequence of reactions for assimilation of N2 and ammonia. The glutamine synthetase in cell-free extracts of the organism is completely sedimented by centrifugation at 140,000 X g for 2 h, is inhibited by L-alanine but not by adenosine 5'-monophosphate, and exhibits two apparent Km values for ammonia (ca. 13 muM and 1 mM).
lacY mutant of Escherichia coli with altered physiology of lactose induction
Authors: Flagg J L , Wilson T H
Published: January 2020

A mutant of Escherichia coli is described that grew on lactose only in the presence of isopropylthiogalactoside. This cell contained a defect in the lacY gene that resulted in the formation of a transport system with a poor affinity for lactose. The inability to grow on lactose alone was due to the failure of induction by this disaccharide. This failure of inducation was presumably due to a defect in lactose accumulation which resulted in significant reduction in the formation of allo-lactose, the true inducer of lac operon. These results are consistent with the view that the capacity to accumulate lactose plays an important physiological role in the induction of the enzymes necessary for its utilization.
Salmonella typhimurium mutants generally defective in chemotaxis
Authors: Collins A L , Stocker B A
Published: January 2020

The mutations of eight chemotaxis-deficient strains of Salmonella typhimurium, including five new mutants in strain LT2, were mapped by P22 transduction in relation to various fla mot deletions in S. abortus-equi. Seven recessive che mutations mapped between motB and flaC: three, all nontumbling, the che region I, adjacent to motB, and four, including one ever-tumbling, in che region II, adjacent to flaC. Mutant che-107, never-tumbling and dominant to wild type, mapped at flaAII, other mutations of which cause either absence of flagella or lack of locomotor function. We surmise that gene flaAII specifies a protein that polymerizes to form an essential component of the basal apparatus (so that absence of gene product prevents formation of flagela); that a component built up from certain mutationally altered proteins cannot transmit (or generate) active rotation of the hook and flagellum, and so causes the Mot (paralysis) phenyotype; and that a component built up from protein with the che-107 alteration permits only counterclockwise rotation, so that the tumble, normally produced by transient clockwise rotation, cannot be effected.
Reduced nicotinamide adenine dinucleotide oxidase activity in membranes and cytoplasm of Acholeplasma laidlawii and Mycoplasma mycoides subsp. capri
Authors: Larraga V , Razin S
Published: January 2020

The properties of the membrane-bound reduced nicotinamide adenine dinucleotide (NADH) oxidase of Acholeplasma laidlawii were compared with those of the corresponding cytoplasmic activity of Mycoplasma mycoides subsp. capri. The striking differences in pH optima, susceptibility to inhibitors and detergents, and heat inactivation between the NADH oxidase activity, with oxygen as an electron acceptor, and the NADH oxidoreductase activity, with dichlorophenol indophenol (DCPIP) as an alternate electron acceptor, support the presence of more than one catalytic protein in both the membrane-bound and soluble enzyme systems. The detection of more than one band positive for the NADH-nitroblue tetrazolium oxidoreductase reaction on electrophoresis of either the membranes of A. laidlawii or the cytoplasm of M mycoides subsp. capri also points in the same direction. The membrane-bound enzyme system differed, however, form the soluble one because it had a lower ratio of oxidase activity to oxidoreductase activity, and because it was less susceptible to heat inactivation and more readily incorporated incorporated into reaggregated membranes. In addition, the specific activity of the membrane-bound enzyme system increased as the culture aged, whereas that of the soluble system decreased as the culture aged. It is suggested that the different location in the cell could be responsible for some of the differences between the membrane-bound NADH oxidase activity of A. laidlawii and that found in the cytoplasm of M. mycoides subsp. capri.
Diploidy for a structural gene specifying a major protein of the outer cell envelope membrane from Escherichia coli K-12
Authors: Datta D B , Krämer C , Henning U
Published: January 2020

Homogenotes, heterogenotes, and intergeneric hybrids have been studied that are diploid for the structural gene of a major outer cell envelope membrane protein (protein II) from Escherichia coli. This protein can act as a phage receptor. In wild-type homogenotes, diploidy for the gene did not cause a gene dosage effect. It could be shown with two heterogenotes that both the chromosomal mutant and the episomal wild-type genes are expressed, and in each case more of the mutant than the wild-type protein species was found in the cell envelope. In on case of 21 phage-resistant mutants missing protein II was a trans effect observed of the mutant gene on the expression of the episomal wild type gene. Transfer of E. coli episomes carrying the protein II structural gene into Salmonella typhimurium and Proteus mirabilis resulted in intergeneric hybrids that became sensitive to the relevant phage and harbored the E. coli protein II in their cell envelopes. The results may be taken as suggestive evidence for a simple feedback mechanism for the regulation of synthesis of protein II, and they show that there are no highly specific requirements on protein primary structure for incorporation into an outer cell envelope membrane.
Multiple-carbon-source-limited growth kinetics of a marine coryneform bacterium
Authors: Law A T , Button D K
Published: January 2020

The steady-state growth rate of a marine isolate was related to the concentrations of several carbon and energy source substrates when these substrates limited growth simultaneously in continuous culture. Glucose limitation was characterized by a threshold of 0.21 mg/liter for growth, a half-maximal growth rate at 0.48 mg/liter, U-shaped curves in extractable pool concentration-versus-growth velocity plots, and slow maximal growth rates. Arginine addition reduced the glucose threshold to 0.008 mg/liter, more than doubled the maximal growth rate, and stabilized pool concentrations at low growth rates. Addition of a third substrate, glutamate, caused further reduction of the glucose concentration a steady state. Maximal reduction of the glucose concentration was effected by adding a mixture of 20 amino acids. Steady-state limiting nutrient concentration was dependent on the specific identity of the auxiliary nutrients and on the concentration ratio at which they were supplied. When glucose was supplemented with an equal quantity of an amino acid mixture, the external steady-state glucose remained below 10 mug/liter. When 1 mug of glucose was added to a 2.5-mg/liter amino acid mixture, at least 70% of it was consumed at steady state in spite of the threshold observed. Lack of crossover between metabolic pathways, suggested by the absence of glucose carbon in pool glutamate of arginine-glucose-grown cells, may have been partly responsible for the mixed carbon source stimulation of nutrient accumulation observed. The affinity observed is sufficient to account for normal growth at a total organic substrate concentration of only 0.11 mg/liter when supplied from a suitable mixture.
Effect of entry exclusion on mating aggregates and transconjugants
Authors: Eckerson H W , Reynard A M
Published: January 2020

Mating aggregates during conjugation directed by an F-like R factor in Escherichia coli were measured as the number of Lac+-Lac- sectored colonies present in a mating mixture. There is a high degree of correlation between the concentration of transconjugants produced in a mating mixture and the concentration of mating aggregates observed at several different concentrations of donor and recipient cells. The mating aggregates are sex pilus specific as demonstrated by the ability of donor-specific ribonucleic acid phage MS-2 to decrease both mating aggregates and transconjugants in a mating mixture. During entry exclusion by either a derepressed or a repressed F-like R factor, isogenic to the superinfecting R factor except for a resistance determinant, the number of transconjugants was markedly reduced, but the number of mating aggregates was not decreased. Entry exclusion by F-Gal toward the donor HfrH resembled that of the F-like R factor in that there was a reduction in the number of recombinants but no significant decrease in mating aggregates. These results suggest that entry exclusion inhibits conjugation at a stage after the formation of mating aggregates.
Chemotaxis toward amino acids by Bacillus subtilis
Authors: Ordal G W , Gibson K J
Published: January 2020

Conditions for assaying chemotaxis in Bacillus subtilis are described. The chemotaxis medium we used afforded excellent motility for hours. In it, chemotaxis measured by capillary assays was insensitive to pH between 5.5 and 9, and to temperature between 28 degrees C and 42 degrees C. Chemotaxis was observed toward all 20 common amino acids, with thresholds varying from 3nM for alanine to 0.1 mM for glutamate, in the capillary assay, and from 0.1 muM for alanine to 0.32 mM for glutamate in the microscope assay.
Relation of cell growth and colicin tolerance to vitamin B12 uptake in Escherichia coli
Authors: Kadner R J , Bassford P J
Published: January 2020

The uptake of vitamin B12 was measured in cells of Escherichia coli whose growth had been inhibited by any of a variety of treatments. In all cases, the secondary, energy-dependent phase of B12 uptake was depressed in proportion to the decrease in growth rate, but uptake was constant in cells growing logarithmically at different rates. The depression of B12 uptake activity was independent of the site of cell metabolism affected by the inhibitor or by its effect on cell viability, and was both more rapid and of greater degree than the effects on the uptake of any of the six amino acids tested. The decline was not affected by inhibitors of either cell division or proteolysis and was manifested without any apparent decrease in the surface B12 binding activity. Transport activity was rapidly regained upon reversal of the inhibition of protein synthesis. Prompted by this response, the uptake of B12 was contrasted to the apparent uptake of the E colicins, which share the same outer membrane receptor. Sensitivity to colicin E1, measured by its inhibition of proline uptake, was not affected by growth inhibition by antibiotic treatment. Finally, there was no specific depression of B12 uptake in cells rendered colicin tolerant either by mutation or as a consequence of phage f1 infection.
Genetic inversion in the formation of an Hfr strain from a temperature-sensitive F' gal strain
Authors: Bergquist P L , Jamieson A F
Published: January 2020

An Hfr strain (PB15) that carries a duplicated copy of the galactose operon genes flanking the integrated sex factor is unusually stable since it does not show excision of the repeated deoxyribonucleic acid segment. The right-hand galactose operon is in the normal orientation. Deletion mutations that eliminate the right-hand galactose genes, the sex factor, and some of the left-hand operon have been isolated. Mutants believed to have their left-hand galactose operon inverted were able to be induced for galactose epimerase synthesis by D-fucose but did not show escape synthesis on induction of bacteriophage lambda. Ribonucleic acid specific for the galactose operon was isolated after induction of lysogenic strains presumed to carry the galactose operon in the normal and inverted orientation. Hybridization to the isolated left and right strands of lambdapgal showed that the noninformational strand of the left-hand galactose operon of the deletion mutant of PB15 was transcribed on escape induction. These results show that inversion has occurred.
EcoRI cleavage sites in the argECBH region of the Escherichia coli chromosome
Authors: Devine E A , Moran M C , Jederlinic P J , Mazaitis A J , Vogel H J
Published: January 2020

The EcoRI cleavage of deoxyribonucleic acids (DNAs) from lambdadarg phages, carrying argECBH, has been examined. The phages are derived from the heat-inducible, lysis-defective strain lambda y199, and their bacterial DNA, including argECBH, is derived from Escherichia coli K-12. Such cleavage of the phage DNAs, in each case, produces the D, E, and F segments of lambda. Additionally, these DNAs yield segments, ordered from left to right, of length (in kilobases [kb]) determined by electron microscopy and 0.7% agarose slab gel electrophoresis as follows: lambdadarg13 (ppc argECBH bfe), 13.9, 2.8, 1.5, and 5.6; lambdadarg14 (ppc argECBH), 3.0, 2.0, 17.3, and 6.2; and lambdadarg23 (argECBH), 18.4 and 6.2. For lambdadarg13 sup102 DNA, the segment analogous to the 13.9-kb segment measures 12.2 kb. The direction from left to right corresponds to the clockwise orientation of the E. coli genetic map. The EcoRI segments define five cleavage sites near the arg region of the E. coli chromosome. For each of the DNAs, the arg genes occur on the largest segment produced. The 17.3-kb segment, being entirely bacterial, represents the argECBH-bearing EcoRI segment of the E. coli chromosome. The location of the arg genes was demonstrated electron microscopically in heteroduplex experiments.
Genetic Characterization of the Temperature-Sensitive and Suppression Phenotypes of
Escherichia coli
Mutant N4316
Authors: Herrington M. B. , Ganoza M. C.
Published: January 2020

Escherichia coli
mutant N4316 is temperature sensitive and exhibits temperature-dependent suppression. These phenotypes are due to separate genes, as shown by reversion and mapping studies. The suppressor mutation was mapped and lies near
argF
.
Control of cell division in Escherichia coli: effect of amino acid starvation
Authors: Ron E , Grossman N , Helmstetter C E
Published: January 2020

The effect of amino acid starvation on cell division was studied in cells of Escherichia coli B. In this bacterial strain, deprivation of a required amino acid resulted in synchronous cell division upon restoration of the amino acid. This synchronization was apparently due to a shift forward in the cell cycle during the starvation. As a consequence, the cells divided at a size that was smaller than normal.
Rate of ribonucleic acid chain growth in Mycobacterium tuberculosis H37Rv
Authors: Harshey R M , Ramakrishnan T
Published: January 2020

Two methods were employed to measure the rate of ribonucleic acid (RNA) chain growth in vivo in Mycobacterium tuberculosis H37Rv cultures growing in Sauton medium at 37 degrees C, with a generation time of 10 h. In the first, the bacteria were allowed to assimilate [3H]uracil or [3H]guanine into their RNA for short time periods. The RNA was then extracted and hydrolyzed with alkali, and the radioactivity in the resulting nucleotides and nucleosides was measured. The data obtained by this method allowed the calculation of the individual nucleotide step times during the growth of RNA chains, from which the average rate of RNA chain elongation was estimated to be about 4 nucleotides per s. The second method employed the antibiotic rifampin, which specifically inhibits the initiation of RNA synthesis without interfering with the elongation and completion of nascent RNA chains. Usint this method, the transcription time of the 16S, 23S, and 5S ribosomal RNA genes was estimated to be 7.6 min, which corresponds to a ribosomal RNA chain growth rate of 10 nucleotides per s.