scholarly journals Thermal denaturation in acidic solutions of double-helical ribonucleic acid from virus-like particles found in Penicillium chrysogenum. A spectrophotometric study

1971 ◽  
Vol 125 (2) ◽  
pp. 655-665 ◽  
Author(s):  
R. A. Cox ◽  
K. Kanagalingam ◽  
Elisabeth Sutherland

1. Two species of double-helical RNA isolated from mycelium of Penicillium chrysogenum were titrated with acid at 25°C and 95°C (solvent 0.1m-sodium phosphate buffer). At 25°C denaturation occurred at about pH3. At 95°C in the denatured form cytosine residues titrated as a simple monobasic acid of pK3.9 compared with pK≃2.5 for the native form at 25°C. 2. On thermal denaturation in neutral and acidic solutions one species of RNA (38% rG·rC) ‘melted’ in three distinct stages, equivalent to a mixture of three species, namely one of about 25% rG·rC, another of about 33% rG·rC and a third of about 46% rG·rC: the relative proportions were 0.25:0.35:0.40. 3. On thermal denaturation in acidic solutions the increase in the fraction of ionized cytosine residues concomitant with the ‘melting’ of rG·rC base pair also affects the spectrum especially at 280nm and serves to enhance the contribution of rG·rC base pairs at this wavelength. The increment in ε(P) at 280nm on ‘melting’ an rG·rC base pair approaches 53501·mol−1·cm−1 depending on pH, compared with 33501·mol−1·cm−1 at pH7. In contrast ε(P) at 280nm is scarcely affected by ‘melting’ rA·rU base pairs or by the protonization of adenine residues. 4. Changes in the spectrum of Escherichia coli rRNA on denaturation in acidic solutions were studied to yield the mole fractions of rA·rU and rG·rC base pairs ‘melting’ at particular pH values.

1968 ◽  
Vol 108 (4) ◽  
pp. 599-610 ◽  
Author(s):  
R A Cox ◽  
K Kanagalingam

1. The thermal denaturation of DNA from rat liver was studied spectrophotometrically. In sodium phosphate buffers denaturation led to a single-stranded form having, at 25°, about 25% of the hypochromism of the intact double helix. 2. The hypochromism of the denatured form was the same in 1mm- as in 10mm-sodium phosphate buffer and was scarcely affected by reaction with formaldehyde. The hypochromism was decreased by about 40% in the presence of 8m-urea. 3. The hypochromism of denatured DNA at low ionic strengths was about the same as that of fragments of reticulocyte ribosomal RNA that were too short to form double-helical secondary structure and about the same as that of RNA after reaction with formaldehyde. 4. The spectrum of DNA was slightly affected by the presence of 8m-urea or 4m-guanidinium chloride. The differences in the spectrum of the native and denatured forms of DNA in 0·1m-sodium phosphate buffer, in 8m-urea–10mm-sodium phosphate buffer and in 4m-guanidinium chloride–10mm-sodium phosphate buffer, pH7·6, were similar but not identical. 5. Denatured rat liver DNA appears to have no double-helical character at 25° in 10mm-sodium phosphate buffer, pH7·6; increasing the buffer concentration to 0·1m leads to a more compact form in which about 40% of the residues form base pairs.


1988 ◽  
Vol 252 (2) ◽  
pp. 481-487 ◽  
Author(s):  
R K Tripathi ◽  
C Chaya Devi ◽  
A Ramaiah

1. We have shown that the characteristic lag in cresolase activity of human skin tyrosinase at inhibitory concentration of tyrosine was absent at all pH values studied, i.e. pH 5.2, 5.7, 6.2 and 6.8, if the enzyme solubilized at low pH was used as the source of enzyme, but the same enzyme when dialysed against buffers of various pH values showed linear activity only at pH 5.2 and was not inhibited by excess tyrosine, whereas at higher pH values it exhibited a lag and inhibition by excess tyrosine. 2. However, the enzyme solubilized in buffer/detergent, pH 6.8, when dialysed against buffer of the same pH showed linear activity at pH 5.2 and non-linear activity at pH 6.8. 3. The water/detergent-solubilized enzyme from human skin melanosomes showed linear activity even at inhibitory concentrations of tyrosine at pH 5.2 and 6.8 up to 2 h, but acceleration of rate was observed after 2 h for the enzyme measured at pH 6.8. 4. After dialysis of the water/detergent-solubilized enzyme against double-glass-distilled water, it still exhibits linear activity at inhibitory concentration of tyrosines at pH 6.8 for the first 2 h, but the same enzyme when dialysed against 0.02 M-sodium phosphate buffer, pH 6.8, exhibits negligible activity up to 1/2 h, in contrast with considerable activity before dialysis during the same interval of time, but without any loss of activity at later intervals of incubation time. 5. On the basis of these results, it is concluded that the enzyme exists in at least two interconvertible forms, one without lag and inhibition by excess tyrosine and the other with lag and inhibition by excess tyrosine. These two forms are interconvertible only by gradual change in pH over a period of hours.


2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Masayo Suzuki ◽  
Kazuya Ohtsuki ◽  
Katsuhito Kino ◽  
Teruhiko Kobayashi ◽  
Masayuki Morikawa ◽  
...  

The nucleoside 2,2,4-triamino-5(2H)-oxazolone (Oz) can result from oxidative damage to guanine residues in DNA. Despite differences among the three polymerases (Polβ, KF exo−, and Polη) regarding nucleotide incorporation patterns opposite Oz, all three polymerases can incorporate guanine opposite Oz. Based onab initiocalculations, we proposed a structure for a stable Oz:G base pair. Here, to assess the stability of each Oz-containing base pair (Oz:G, Oz:A, Oz:C, and Oz:T) upon DNA replication, we determined the efficiency of Polβ-, KF exo−-, or Polη-catalyzed primer extension beyond each base pair. With each polymerase, extension beyond Oz:G was more efficient than that beyond Oz:A, Oz:C, or Oz:T. Moreover, thermal denaturation studies revealed that theTmvalue for the duplex containing Oz:G was significantly higher than those obtained for duplexes containing Oz:A, Oz:C, or Oz:T. Therefore, the results fromab initiocalculations along with those from DNA replication assays and thermal denaturation experiments supported the conclusion that Oz:G is the most stable of the Oz-containing base pairs.


1998 ◽  
Vol 51 (11) ◽  
pp. 965 ◽  
Author(s):  
Nancy Guzzo-Pernell ◽  
John M. Lawlor ◽  
Geoffrey W. Tregear ◽  
Jim Haralambidis

2′-Deoxy-N4-phenylcarbamoylcytidine (Pc) was designed for triplex recognition of the cytosine{guanine base pair at physiological pH values to assist in the development of triplex-forming oligomers. Thermal denaturation of a triplex mixture in which the third strand incorporated Pc as a central nucleoside showed that, at pH 6·1, Pc binds selectively—though weakly—to the target cytosine—guanine and also to guanine—cytosine; but the gel mobilities of the same mixtures indicate the possibility of specificity for the target cytosine-guanine base pair at pH values higher than 6·1. The result may provide leads useful in the development of improved triplex-forming oligomers.


Akademos ◽  
2021 ◽  
Vol 60 (1) ◽  
pp. 51-56
Author(s):  
Elena Tofan ◽  
◽  
Natalia Chiselita ◽  
Oleg Chiselita ◽  
Alina Besliu ◽  
...  

Currently, special attention is paid to the use of industrial by-products, in particular those obtained in huge quantities in the brewing and wine making process, as a source for the production of natural preparations with high biological value. In this study are presented results related to the recovery of beer yeast biomass from the sediments of the beer industry, by obtaining lipid extracts with valuable biochemical composition. Thus, applying the optimized autolysis method, with the use of sodium phosphate buffer at 45°C for the destruction of the cell wall and fractional extraction with hydric, alkaline and acidic solutions, various liquid and solid fractions were obtained from the biomass of brewer’s yeast with varied lipid content. So, in order to optimize the waste processing process of the beer industry and the complex recovery of yeast biomass, it is reasonable to include the lipid fraction extraction stage in the technological flow, after obtaining protein and mannoprotein extracts, which will be useful and as an additional step of purification of the β-glucan fraction.


Author(s):  
G.J. Spector ◽  
C.D. Carr ◽  
I. Kaufman Arenberg ◽  
R.H. Maisel

All studies on primary neural degeneration in the cochlea have evaluated the end stages of degeneration or the indiscriminate destruction of both sensory cells and cochlear neurons. We have developed a model which selectively simulates the dystrophic changes denoting cochlear neural degeneration while sparing the cochlear hair cells. Such a model can be used to define more precisely the mechanism of presbycusis or the hearing loss in aging man.Twenty-two pigmented guinea pigs (200-250 gm) were perfused by the perilymphatic route as live preparations using fluorocitrate in various concentrations (15-250 ug/cc) and at different incubation times (5-150 minutes). The barium salt of DL fluorocitrate, (C6H4O7F)2Ba3, was reacted with 1.0N sulfuric acid to precipitate the barium as a sulfate. The perfusion medium was prepared, just prior to use, as follows: sodium phosphate buffer 0.2M, pH 7.4 = 9cc; fluorocitrate = 15-200 mg/cc; and sucrose = 0.2M.


1991 ◽  
Vol 66 (04) ◽  
pp. 500-504 ◽  
Author(s):  
H Peretz ◽  
U Seligsohn ◽  
E Zwang ◽  
B S Coller ◽  
P J Newman

SummarySevere Glanzmann's thrombasthenia is relatively frequent in Iraqi-Jews and Arabs residing in Israel. We have recently described the mutations responsible for the disease in Iraqi-Jews – an 11 base pair deletion in exon 12 of the glycoprotein IIIa gene, and in Arabs – a 13 base pair deletion at the AG acceptor splice site of exon 4 on the glycoprotein IIb gene. In this communication we show that the Iraqi-Jewish mutation can be identified directly by polymerase chain reaction and gel electrophoresis. With specially designed oligonucleotide primers encompassing the mutation site, an 80 base pair segment amplified in healthy controls was clearly distinguished from the 69 base pair segment produced in patients. Patients from 11 unrelated Iraqi-Jewish families had the same mutation. The Arab mutation was identified by first amplifying a DNA segment consisting of 312 base pairs in controls and of 299 base pairs in patients, and then digestion by a restriction enzyme Stu-1, which recognizes a site that is absent in the mutant gene. In controls the 312 bp segment was digested into 235 and 77 bp fragments, while in patients there was no change in the size of the amplified 299 bp segment. The mutation was found in patients from 3 out of 5 unrelated Arab families. Both Iraqi-Jewish and Arab mutations were detectable in DNA extracted from blood and urine samples. The described simple methods of identifying the mutations should be useful for detection of the numerous potential carriers among the affected kindreds and for prenatal diagnosis using DNA extracted from chorionic villi samples.


2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Sung-Wook Yun ◽  
Chan Yu

Decades of mining in South Korea have resulted in the contamination of large amounts of soil by metals. The most feasible approach to site restoration requires the use of a stabilization agent to reduce metal mobility. This study examined the leaching characteristics of limestone used as a stabilization agent when subjected to solutions of differing pH. In a laboratory-scale column test, solutions with pH values of 3.5, 4.6, and 5.6, representing acidic to nonacidic rainfall, were applied to soil mixed with limestone. Test results indicate that metal components can be released with the addition of acidic solutions, even if the soil is highly alkaline. Cd and Zn, in particular, exhibited abrupt or continuous leaching when exposed to acid solutions, indicating the potential for contamination of water systems as metal-laden soils are exposed to the slightly acidic rainfall typical of South Korea. Treatment using stabilization agents such as limestone may reduce leaching of metals from the contaminated soil. Stabilizing metal-contaminated farmland is an economical and feasible way to reduce pollutants around abandoned metal mines.


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