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Biomedical subjects

A Hinkkanen

Publications and source records attributed to A Hinkkanen.

23 records · Page 2Linked to original sources
PubMed1983

Enzymatic determination of several D-amino acids using luminol-mediated chemiluminescence.

A method for the quantitative determination of several D-amino acids in the range of 0.05-1 nmol per assay (0.25-5 microM) is described. It is insensitive to the presence of excesses of the respective L-amino acids. The assay system employs D-amino-acid oxidase (hog kidney), peroxidase (horse radish) and luminol; the total photon output elicited by the oxidation of the D-amino acids is determined. The different reactivity of individual D-amino acids with D-amino-acid oxidase limits the applicability of the assay. Indications for the usefulness of immobilized enzymes in D-amino-acid analysers are also given.

Amino Acids↗
PubMed1983

Purification of the flavoproteins 6-hydroxy-D- and 6-hydroxy-L-nicotine oxidase using hydrophobic affinity chromatography.

A systematic study of the homologous series of omega-aminoalkyl-agaroses revealed differences in the affinities of 6-hydroxy-D- and 6-hydroxy-L-nicotine oxidase. In contrast to supports with nonpolar alkyl chains, omega-aminoalkyl-agarose showed high affinity towards the L-specific enzyme, while the D-specific oxidase was bound most firmly by omega-aminododecyl-agarose. 6-Hydroxy-L-nicotine oxidase could be desorbed by 1.3M NaCl only in the presence of the substrate L-6-hydroxynicotine. Using the omega-aminoalkyl-agarose, a complete separation of the enantiozymes was accomplished and an efficient purification procedure for both oxidases established.

Arthrobacter↗
PubMedDate not supplied

Oligonucleotide fingerprinting with (CAC)5: nonradioactive in-gel hybridization and isolation of individual hypervariable loci.

The first topic to be treated in this paper is the nonradioactive DNA fingerprinting by means of in-gel hybridization with digoxigenated (CAC)5. Besides the fact that time-consuming Southern blotting can be avoided, the dried agarose is an excellent matrix to produce background-free nonradioactive DNA fingerprints. There is no tendency of either the oligonucleotide probe or the antibody towards unspecific binding to the dried agarose. Prehybridization and blocking steps are therefore superfluous. Furthermore, we will discuss what effect the degree of crosslinking of the antibody-enzyme conjugates has. The second topic concerns the isolation and characterization of locus-specific probes from a human (CAC)5 fingerprint. The isolation and characterization of one variable probe, by screening complete genomic libraries, is described and discussed. This probe is compared to a hypervariable single-copy probe, isolated from a size-enriched genomic library. The sequence of the repeat flanking locus-specific probe is presented and a semi-specific, adaptor-mediated polymerase chain reaction was designed to amplify (CAC)n/(GTG)n flanking sequences.

Base Sequence↗
PubMedDate not supplied

Highly instable (GATA)n-containing sequences of the mouse during the cloning process.

Mouse DNA fingerprints were obtained by HaeIII digestion of genomic DNA and in-gel hybridization with the (GATA)4 oligonucleotide probe. In order to obtain locus-specific probes that hybridize with only one fragment of the (GATA)4 DNA fingerprint, a genomic library of size-selected inserts was constructed using a system of direct subcloning from the phage clones. During the cloning procedure, the phage as well as the plasmid insert DNAs changed primarily within their repetitive DNA but also within adjacent nonrepetitive sequences, as was demonstrated for several clones by in-gel hybridization with the (GATA)4 probe as well as by sequence analysis. Isolated subclones varied within their (GATA)n repeats, resulting in different insert lengths. Several "metastable" as well as stable (GATA)4-positive subclones could be isolated. Also, vector sequences were affected by alterations during the cloning process. These phenomena are discussed within the context of possible mechanisms for cloning artifacts.

Animals↗