Mercuration nucleic acid

Hopman, A.H.N., Wiegant, J., Tesser, G.I., and Van Duijn, R (1986) A nonradioactive in situ hybridization method based on mercurated nucleic acid probes and sulfhydryl-hapten hgands. Nucleic Acids Res. 14, 6471-6488. 

The hybridization potential of a probe remains intact after mercuration. Sulfhydryl compounds bind strongly to mercurated nucleic acids and a variety of sulfhydryl-(primary label) ligands can be introduced in the complex after hybridization. Simpson (1961) observed a typical association constant of mercury for sulfhydryl compounds of 10 M but the mercury-sulfhydryl bond in polynucleotide complexes may be less stable (Hopman et al., 1986a). An interesting application... 

Mercurated nucleic acids for sulfhydryl-hapten ligands... 

Other methods that are related to affinity chromatography include hydrophobic interaction chromatography and thiophilic adsorption. The former is based on the interactions of proteins, peptides, and nucleic acids with short nonpolar chains on a support. This was first described in 1972 [113,114] following work that examined the role of spacer arms on the nonspecific adsorption of affinity columns [114]. Thiophilic adsorption, also known as covalent or chemisorption chromatography, makes use of immobilized thiol groups for solute retention [115]. Applications of this method include the analysis of sulfhydryl-containing peptides or proteins and mercurated polynucleotides [116]. 

Azauracil [1,2,4-triazine-3,5(2,4)-dione] inhibits the growth of various micro-organisms. When grown in the presence of 6-azauracil- -C, Streptococcus jaecalis accumulates radioactive metabolites in the acid-soluble fraction of the cells. A major metabolite is D-ribofuranosyl-6-aza-uracil. This material is identical with material prepared by condensing tri-O-benzoyl-D-ribofuranosyl chloride with the mercuric derivative of 6-azauracil, followed by debenzoylation. A second major metaboUte was tentatively shown to be D-ribosyl-6-azauracil 5-phosphate. Bacteria develop resistance against 6-azauracil and its D-ribosyl derivative. Resistant Streptococcus faecalis will not convert 6-azauracil to its D-ribosyl derivative or to other bound forms, and the bacterium has also lost the ability to incorporate uracil into the nucleic acids of its cells. 

There is little or no reaction with adenine, thymine or guanine bases. Polymer mercuration is, in contrast to mononucleotides, influenced by the ionic strength of the reaction mixture the reaction rate is higher at a lower ionic strength. Residues in ss or ds polymers react at essentially the same rate. Two simultaneous reactions occur a noncovalent complexing reaction and a slower covalent introduction of mercury at the 5 position. It is important to remove the complex-bound mercury ions from the nucleic acids because they interfere with the annealing. Both cyanide and iodide are effective in... 

Analytical Techniques and Physical Methods.— The mapping of oligonucleotides and nucleic acid digests on cellulose or cellulose-polyethyleneimine has been described recently, and columns of mercurated dextran or dihydroxyborylcellulose have been used to fractionate nucleotide mixtures. Electrophoresis on polyacrylamide gels has been advocated as a rapid method for desalting and fractionating mixtures of oligonucleotides. ... 

Van Gennip in Sherma and Fried (8) also reviewed the use of colorimetric agents for the detection of nucleic acid and their derivatives. Common reagents include mercuric acetate (purine and... 

---