scholarly journals CHARACTERIZATION OF T4 MUTANTS THAT PARTIALLY SUPPRESS THE INABILITY OF T4rII TO GROW IN LAMBDA LYSOGENS

Genetics ◽  
1976 ◽  
Vol 83 (3) ◽  
pp. 477-487
Author(s):  
Theodore Homyk ◽  
Angel Rodriguez ◽  
Jon Weil

ABSTRACT In the course of isolating viable T4 deletions that affect plaque morphology (Homyk and Weil 1974), two closely linked point mutants, sip1 and sip2, were obtained. They map between genes t and 52, cause a reduction in plaque size and burst size, and partially suppress the lethality of rII mutants for growth in lambda lysogens. These characteristics demonstrate that sip1 and sip2 are similar to mutants previously reported by Freedman and Brenner (1972). In addition, D. Hall (personal communication) has shown that sip1 and sip2 are similar to the mutant farP85, which affects the regulation of a number of early genes (Chace and Hall 1975).—Sip suppression of rII mutants can be demonstrated in one-step growth experiments, even when both rII genes are completely deleted. This indicates that sip mutants do not simply reduce the level of rII gene products required for growth in a lambda lysogen. Instead, they alter the growth cycle so as to partially circumvent the need for any rII products.—Mutations at two other sites, designated L1 and L2, reverse the poor phage growth caused by sip and, in the one case tested, reverse the rII-suppressing ability of sip.

1984 ◽  
Vol 30 (5) ◽  
pp. 521-525 ◽  
Author(s):  
Philippe de Lajudie ◽  
Didier Bogusz

Two rhizobiophages, RS1 and RS2, were isolated in Senegal from a soil sample and dry stem nodules of Sesbania rostrata, a tropical legume that is infected by two categories of Rhizobium strains: "stem strains," which nodulate both roots and stems (type strain, ORS571), and "root strains," which induce effective nodules only on roots. Both phages were found to have a host range restricted to ORS571; all root strains were found to be resistant. By electron microscopy, phage RS1 showed an hexagonal head 63 nm wide and a tail 87 nm long; phage RS2 revealed an hexagonal head 60 nm wide. Characterization of phage growth cycle by one-step growth experiments showed that the latent period was ca. 75 min for RS1 and ca. 4 h for RS2, that the rise period lasted ca. 2 h for both RS1 and RS2, and that the average burst size was ca. 100 for RS1 and 130 for RS2. Temperature denaturation occurred at 60–65 °C (RS1) and 45–50 °C (RS2). Serum neutralization tests revealed that the phages were not serologically related. In contrast to RS1, RS2 appeared to be temperate, since stable lysogens were isolated.


2008 ◽  
Vol 19 (1) ◽  
pp. 83 ◽  
Author(s):  
Neeraja Sankaran

The demonstration of the one-step growth pattern of the bacteriophages is generally regarded as the key evidence that bacteriophages were viruses rather than enzymes of bacterial origin, a matter of considerable debate among scientists since the bacteriophage was first described in 1917. While the credit for this demonstration is usually accorded to a 1939 paper on phage growth by Emory Ellis and Max Delbr�ck, closer scrutiny of phage research conducted in the intervening two decades reveals that these papers did not present a new idea, but rather extended and refined a line of investigation about the phages that had its conceptual antecedents in the earlier work. Of particular note is the work of the Australian, Frank Macfarlane Burnet, during the late 1920s and early 1930s. Burnet's work also furnished other important reasons besides one-step growth—derived from experiments on lysogeny—for favouring the virus theoryand discarding the enzyme theory of phage. This paper examines Burnet's contributions towards understanding of the nature of phage and makes the case that it was a tacit acceptance of the evidence and arguments that he presented that allowed Ellis and Delbr�ck to make assumptions about the bacteriophage, presented as fact in their papers.


1973 ◽  
Vol 11 (1) ◽  
pp. 98-106 ◽  
Author(s):  
Simone Manteuil ◽  
Jacqueline Pages ◽  
Dominique Stehelin ◽  
Marc Girard

1968 ◽  
Vol 14 (4) ◽  
pp. 397-402 ◽  
Author(s):  
Marvin D. Brailsford ◽  
Paul A. Hartman

Streptococcus durans bacteriophages were isolated from bovine rumen fluid, feed lot soil, bovine fecal samples, and lysogenic strains of S. durans. The presence of different phage strains was suggested by antiserum neutralization tests and one-step growth characteristics, whereby the bacteriophages were placed into three distinct, but serologically related, groups. These phages were further characterized by plaque morphology, phage morphology, adsorption kinetics, and pH and temperature effects.


2007 ◽  
Vol 69 (1) ◽  
pp. 174-179 ◽  
Author(s):  
Rimantas Daugelavičius ◽  
Aušra Gaidelytė ◽  
Virginija Cvirkaitė-Krupovič ◽  
Dennis H. Bamford

2021 ◽  
Vol 9 (1) ◽  
pp. 152
Author(s):  
Carly M. Davis ◽  
Jaclyn G. McCutcheon ◽  
Jonathan J. Dennis

Pseudomonas aeruginosa is a pernicious bacterial pathogen that is difficult to treat because of high levels of antibiotic resistance. A promising alternative treatment option for such bacteria is the application of bacteriophages; the correct combination of phages plus antibiotics can produce synergistic inhibitory effects. In this study, we describe morphological changes induced by sub-MIC levels of the antibiotic aztreonam lysine (AzLys) on P. aeruginosa PA01, which may in part explain the observed phage–antibiotic synergy (PAS). One-step growth curves for phage E79 showed increased adsorption rates, decreased infection latency, accelerated time to lysis and a minor reduction in burst size. Phage E79 plus AzLys PAS was also able to significantly reduce P. aeruginosa biofilm growth over 3-fold as compared to phage treatment alone. Sub-inhibitory AzLys-induced filamentation of P. aeruginosa cells resulted in loss of twitching motility and a reduction in swimming motility, likely due to a reduction in the number of polar Type IV pili and flagella, respectively, on the filamented cell surfaces. Phage phiKZ, which uses Type IV pili as a receptor, did not exhibit increased activity with AzLys at lower sub-inhibitory levels, but still produced phage–antibiotic synergistic killing with sub-inhibitory AzLys. A one-step growth curve indicates that phiKZ in the presence of AzLys also exhibits a decreased infection latency and moderately undergoes accelerated time to lysis. In contrast to prior PAS studies demonstrating that phages undergo delayed time to lysis with cell filamentation, these PAS results show that phages undergo accelerated time to lysis, which therefore suggests that PAS is dependent upon multiple factors, including the type of phages and antibiotics used, and the bacterial host being tested.


2020 ◽  
Vol 104 (16) ◽  
pp. 7023-7035
Author(s):  
Samuel Sutiono ◽  
Bettina Siebers ◽  
Volker Sieber

Abstract2-keto-3-L-arabinonate dehydratase (L-KdpD) and 2-keto-3-D-xylonate dehydratase (D-KdpD) are the third enzymes in the Weimberg pathway catalyzing the dehydration of respective 2-keto-3-deoxy sugar acids (KDP) to α-ketoglutaric semialdehyde (KGSA). The Weimberg pathway has been explored recently with respect to the synthesis of chemicals from L-arabinose and D-xylose. However, only limited work has been done toward characterizing these two enzymes. In this work, several new L-KdpDs and D-KdpDs were cloned and heterologously expressed in Escherichia coli. Following kinetic characterizations and kinetic stability studies, the L-KdpD from Cupriavidus necator (CnL-KdpD) and D-KdpD from Pseudomonas putida (PpD-KdpD) appeared to be the most promising variants from each enzyme class. Magnesium had no effect on CnL-KdpD, whereas increased activity and stability were observed for PpD-KdpD in the presence of Mg2+. Furthermore, CnL-KdpD was not inhibited in the presence of L-arabinose and L-arabinonate, whereas PpD-KdpD was inhibited with D-xylonate (I50 of 75 mM), but not with D-xylose. Both enzymes were shown to be highly active in the one-step conversions of L-KDP and D-KDP. CnL-KdpD converted > 95% of 500 mM L-KDP to KGSA in the first 2 h while PpD-KdpD converted > 90% of 500 mM D-KDP after 4 h. Both enzymes in combination were able to convert 83% of a racemic mixture of D,L-KDP (500 mM) after 4 h, with both enzymes being specific toward the respective stereoisomer. Key points• L-KdpDs and D-KdpDs are specific toward L- and D-KDP, respectively.• Mg2+affected activity and stabilities of D-KdpDs, but not of L-KdpDs.• CnL-KdpD and PpD-KdpD converted 0.5 M of each KDP isomer reaching 95 and 90% yield.• Both enzymes in combination converted 0.5 M racemic D,L-KDP reaching 83% yield.


2010 ◽  
Vol 3 (6) ◽  
pp. 5645-5670
Author(s):  
M. Antón ◽  
J. E. Gil ◽  
A. Cazorla ◽  
J. M. Vilaplana ◽  
F. J. Olmo ◽  
...  

Abstract. The ultraviolet (UV) index is the variable most commonly used to inform the general public about the levels and potential harmful effects of UV radiation incident at Earth's surface. This variable is derived from the output signal of the UV radiometers applying conversion factors obtained by calibration methods. This paper focused on the influence of the use of two of these methods (called one-step and two-steps methods) on the resulting experimental UV Index (UVI) as measured by a YES UVB-1 radiometer located in a midlatitude station, Granada (Spain) for the period 2006–2009. In addition, it is also analyzed the difference with the UVI values obtained when the calibration factors provided by the manufacturer are used. For this goal, the detailed characterization of the UVB-1 radiometer obtained in the first Spanish calibration campaign of broadband UV radiometers at the "El Arenosillo" INTA station in 2007 is used. In addition, modeled UVI data derived from the LibRadtran/UVSPEC radiative transfer code are compared with the experimental values recorded at Granada for cloud-free conditions. The absolute mean differences between the measured and modeled UVI data at Granada are around 5% using the one-step and two-steps calibration methods. This result indicates the excellent performance of these two techniques for obtaining UVI data from the UVB-1 radiometer. In contrast, the application of the calibration factor supplied by the manufacturer produces a high overestimation (~14%) of the UVI values. This fact generates unreliable alarming high UVI data in summer when the manufacturer's factor is used. Thus, days with an extreme erythemal risk (UVI higher than 10) increase up to 46% of all cases measured between May and September at Granada when the manufacturer's factor is applied. This percentage is reduced to a more reliable value of 3% when the conversion factors obtained with the two-steps calibration method are used. All these results report about the need of a sound calibration of the broadband UV instruments in order to obtain reliable measurements.


1950 ◽  
Vol 34 (2) ◽  
pp. 231-250 ◽  
Author(s):  
Winston H. Price

1. Four strains of Staphylococcus muscae have been isolated which differ in their growth rates and phage syntheses in Fildes' synthetic medium. 2. Two of the strains when singly infected cannot release phage in Fildes' synthetic medium unless a substance present in certain acid-hydrolyzed proteins is added to the medium. One of these strains also requires other substance(s) present in acid-hydrolyzed proteins in order to grow in Fildes' medium. 3. The two strains which do not require the addition of the phage-stimulating factor have been found either to synthesize this substance, or one similar to it. One of these strains will not grow in Fildes' medium unless substance(s) present in acid-hydrolyzed proteins is added to the medium. 4. The purified acid-hydrolyzed protein factor necessary for virus liberation does not affect the multiplication rate of uninfected S. muscae cells in Fildes' synthetic medium. 5. The substance is not needed for the adsorption or the invasion of the host cell by the virus. In the absence of the factor, the virus is adsorbed to the cell and "kills" it. 6. An analysis carried out by means of the one-step growth curve technique has indicated that the substance is not concerned simply with the mechanism of virus release, but is necessary for some initial stage in virus synthesis. 7. With one bacterial strain not requiring the AHPF, aspartic acid had to be present at least during the minimum latent period for the cell to form virus. 8. In the absence of aspartic acid, the virus was adsorbed to the cell and killed it, but no virus was released from singly infected bacteria. 9. If the cells were grown in a medium containing aspartic acid and then resuspended in the medium minus aspartic acid, no virus was released, although such cells contained at least two times the amount of aspartic acid necessary for the burst size in the complete medium. 10. Aspartic acid, a constituent of the virus particle, appears from an analysis of one-step growth curves to take part in the initial phase of phage synthesis. 11. The effect of amino acids on virus formation is discussed in relation to the time sequence of virus protein and desoxyribonucleic acid synthesis.


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