Selective Staining of Nucleic Acids with Indium

Author(s):  
W. G. Aldridge ◽  
J. R. Coleman

Heavy metal compounds are used to increase contrast in electron microscope specimens by virtue of strong electron scattering properties. Chemical properties of heavy metal atoms cause them to discriminate amongst available ligands, a property useful for biological specimens. Decreasing available specimen ligand types produces a narrow reaction spectrum. Taking advantage of both these controlling factors, one can produce a very narrow reaction spectrum and thus high selectivity. This approach was utilized in the development of methods for the use of indium as an “electron stain” for nucleic acids. Here selectivity is greatly enhanced by reduction and acylation reactions which precede exposure to indium trichloride dissolved in anhydrous acetone. Nucleic acid phosphate is the predominant binding site but some non-nucleic acid ligands still bind indium. To further increase selectivity and provide greater contrast (thus lower detection limits), more efficient blocking reactions of broader chemical reactivity plus techniques to increase the amount of heavy metal bound to nucleic acid were sought.

Life ◽  
2019 ◽  
Vol 9 (3) ◽  
pp. 59 ◽  
Author(s):  
Rong Ni ◽  
Ruilu Feng ◽  
Ying Chau

The discovery of the genetic roots of various human diseases has motivated the exploration of different exogenous nucleic acids as therapeutic agents to treat these genetic disorders (inherited or acquired). However, the physicochemical properties of nucleic acids render them liable to degradation and also restrict their cellular entrance and gene translation/inhibition at the correct cellular location. Therefore, gene condensation/protection and guided intracellular trafficking are necessary for exogenous nucleic acids to function inside cells. Diversified cationic formulation materials, including natural and synthetic lipids, polymers, and proteins/peptides, have been developed to facilitate the intracellular transportation of exogenous nucleic acids. The chemical properties of different formulation materials determine their special features for nucleic acid delivery, so understanding the property–function correlation of the formulation materials will inspire the development of next-generation gene delivery carriers. Therefore, in this review, we focus on the chemical properties of different types of formulation materials and discuss how these formulation materials function as protectors and cellular pathfinders for nucleic acids, bringing them to their destination by overcoming different cellular barriers.


Author(s):  
A.-C. Dock-Bregeon ◽  
D. Moras

At first glance crystallizing nucleic acids poses the same problems as crystallizing proteins since most of the variables to investigate are alike. It is thus astonishing that crystallization data banks (1) that describe so many successful protein crystallizations are so poor in information on nucleic acids. This relies on the physico-chemical and biochemical characteristics of nucleic acids distinguishing them from proteins. The aim of this chapter is to underline features explaining the difficulties often encountered in nucleic acid crystallization and to discuss strategies that could help to crystallize them more readily, either as free molecules or as complexes with proteins. Other general principles, in particular for RNA crystallization, are discussed in ref. 2. Among natural nucleic acids only the smaller ones provide good candidates for successful crystallizations. Large DNAs or RNAs can a priori be excluded because of their flexibility that generates conformational heterogeneity not compatible with crystallization. Thus the smaller RNAs with more compact structures (with 75-120 nt), especially transfer RNAs (tRNAs), but also 5S RNA, were the first natural nucleic acids to be crystallized (3, 4). At present attempts are being made with other RNA systems, such as ribozymes and introns, fragments of mRNA, viroids, viral and other tRNA-like RNAs, SELEX-evolved RNAs, and crystallization successes leading to X-ray structure determinations were reported for RNA domains of up to 160 nt long, with the resolution of the P4-P6 domain of the self-splicing Tetrahymena intron (5). The recent excitement in nucleic acid crystallography, and particularly in RNA crystallography, have partly been due to technological improvements in the preparation methods of the molecules. Advances in oligonucleotide chemical synthesis provide opportunity for making large amounts of pure desoxyribo- and more recently of ribo-oligomers of any desired sequence. This led to the crystallization of a number of DNA and RNA fragments and was followed by the co-crystallization of complexes between proteins and such synthetic fragments. Transcription methods of RNAs from synthetic DNA templates were also essential for rejuvenating the structural biology of RNAs. In the case of complexes of proteins with RNAs, the main difficulty was to purify large quantities of homogeneous biological material with well defined physico-chemical properties.


Author(s):  
Dimitrij Lang

The success of the protein monolayer technique for electron microscopy of individual DNA molecules is based on the prevention of aggregation and orientation of the molecules during drying on specimen grids. DNA adsorbs first to a surface-denatured, insoluble cytochrome c monolayer which is then transferred to grids, without major distortion, by touching. Fig. 1 shows three basic procedures which, modified or not, permit the study of various important properties of nucleic acids, either in concert with other methods or exclusively:1) Molecular weights relative to DNA standards as well as number distributions of molecular weights can be obtained from contour length measurements with a sample standard deviation between 1 and 4%.


Author(s):  
Stephen D. Jett

The electrophoresis gel mobility shift assay is a popular method for the study of protein-nucleic acid interactions. The binding of proteins to DNA is characterized by a reduction in the electrophoretic mobility of the nucleic acid. Binding affinity, stoichiometry, and kinetics can be obtained from such assays; however, it is often desirable to image the various species in the gel bands using TEM. Present methods for isolation of nucleoproteins from gel bands are inefficient and often destroy the native structure of the complexes. We have developed a technique, called “snapshot blotting,” by which nucleic acids and nucleoprotein complexes in electrophoresis gels can be electrophoretically transferred directly onto carbon-coated grids for TEM imaging.


2021 ◽  
Vol 23 (1) ◽  
pp. 219-228
Author(s):  
Nabanita Saikia ◽  
Mohamed Taha ◽  
Ravindra Pandey

The rational design of self-assembled nanobio-molecular hybrids of peptide nucleic acids with single-wall nanotubes rely on understanding how biomolecules recognize and mediate intermolecular interactions with the nanomaterial's surface.


2002 ◽  
Vol 56 (11) ◽  
pp. 483-488
Author(s):  
Sasa Jovanic ◽  
Dragoslav Stoiljkovic ◽  
Ivanka Popovic

The contamination of important synthetic (surface unmodified) polymers by various heavy metal compounds (such as copper, manganese and lead) in aqueous medium was investigated in this study. The influence of the pH of the aqueous medium, temperature and metal type on contamination was investigated during a 10 day period. It was found that increasing pH contributed to higher polymer contamination (at higher pH 100 times for copper and up to 400 times for lead), as well as contact with easily penetrable substances. Increasing temperature decreased contamination by the metal compound for PELD and PET which was not the case for PEHD and PR.


1996 ◽  
Vol 52 (3) ◽  
pp. 414-422 ◽  
Author(s):  
E. N. Maslen ◽  
V. A. Streltsov ◽  
N. Ishizawa

Structure factors for small synthetic crystals of the C-type rare earth (RE) sesquioxides Y2O3, Dy2O3 and Ho2O3 were measured with focused λ = 0.7000 (2) Å, synchrotron X-radiation, and for Ho2O3 were re-measured with an MoKα (λ = 0.71073 Å) source. Approximate symmetry in the deformation electron density (Δρ) around a RE atom with pseudo-octahedral O coordination matches the cation geometry. Interactions between heavy metal atoms have a pronounced effect on the Δρ map. The electron-density symmetry around a second RE atom is also perturbed significantly by cation–anion interactions. The compounds magnetic properties reflect this complexity. Space group Ia{\bar 3}, cubic, Z = 16, T = 293 K: Y2O3, Mr = 225.82, a = 10.5981 (7) Å, V = 1190.4 (2) Å3, Dx = 5.040 Mg m−3, μ 0.7 = 37.01 mm−1, F(000) = 1632, R = 0.067, wR = 0.067, S = 9.0 (2) for 1098 unique reflections; Dy2O3, Mr = 373.00, a = 10.6706 (7) Å, V = 1215.0 (2) Å3, Dx = 8.156 Mg m−3, μ 0.7 = 44.84 mm−1, F(000) = 2496, R = 0.056, wR = 0.051, S = 7.5 (2) for 1113 unique reflections; Ho2O3, Mr = 377.86, a = 10.606 (2) Å, V = 1193.0 (7) Å3, Dx = 8.415 Mg m−3, μ 0.7 = 48.51 mm−1 F(000) = 2528, R = 0.072, wR = 0.045, S = 9.2 (2) for 1098 unique reflections of the synchrotron data set.


2021 ◽  
Vol 11 (15) ◽  
pp. 7099
Author(s):  
Inkyeong Moon ◽  
Honghyun Kim ◽  
Sangjo Jeong ◽  
Hyungjin Choi ◽  
Jungtae Park ◽  
...  

In this study, the geochemical properties of heavy metal-contaminated soils from a Korean military shooting range were analyzed. The chemical behavior of heavy metals was determined by analyzing the soil pH, heavy metal concentration, mineral composition, and Pb isotopes. In total, 24 soil samples were collected from a Korean military shooting range. The soil samples consist of quartz, albite, microcline, muscovite/illite, kaolinite, chlorite, and calcite. Lead minerals, such as hydrocerussite and anglesite, which are indicative of a transformation into secondary mineral phases, were not observed. All soils were strongly contaminated with Pb with minor concentrations of Cu, Ni, Cd, and Zn. Arsenic was rarely detected. The obtained results are indicated that the soils from the shooting range are contaminated with heavy metals and have evidences of different degree of anthropogenic Pb sources. This study is crucial for the evaluation of heavy metal-contaminated soils in shooting ranges and their environmental effect as well as for the establishment of management strategies for the mitigation of environmental risks.


Sign in / Sign up

Export Citation Format

Share Document