Biomedical subjects

E Kejnovský

Publications and source records attributed to E Kejnovský.

5 recordsLinked to original sources

PubMed2001

Heterogeneity of rDNA distribution and genome size in Silene spp.

Genus Silene L. (Caryophyllaceae) contains about 700 species divided into 44 sections. According to recent taxonomic classification this genus also includes taxa previously classified in genera Lychnis and Melandrium. In this work, four Silene species belonging to different sections were studied: S. latifolia (syn. Melandrium album, Section Elisanthe), S. vulgaris (Inflatae), S. pendula (Erectorefractae), and S. chalcedonica (syn. Lychnis chalcedonica, Lychnidiformes). Flow cytometric analysis revealed a genome size of 2.25 and 2.35 pg/2C for S. vulgaris and S. pendula and of 5.73 and 6.59 pg/2C for S. latifolia and S. chalcedonica. All four species have the same chromosome number including the pair of sex chromosomes of the dioecious S. latifolia (2n = 2x = 24). Double target fluorescence in-situ hybridization revealed the chromosomal locations of 25S rDNA and 5S rDNA. A marked variation in number and localization of rDNA loci but no correlation between the numbers of rDNA clusters and genome size was found. FISH and genome size data indicate that nuclear genomes of Silene species are highly diversified as a result of numerous DNA amplifications and translocations.

Cell Nucleolus↗

PubMed2001

Localization of male-specifically expressed MROS genes of Silene latifolia by PCR on flow-sorted sex chromosomes and autosomes.

The dioecious white campion Silene latifolia (syn. Melandrium album) has heteromorphic sex chromosomes, XX in females and XY in males, that are larger than the autosomes and enable their separation by flow sorting. The group of MROS genes, the first male-specifically expressed genes in dioecious plants, was recently identified in S. latifolia. To localize the MROS genes, we used the flow-sorted X chromosomes and autosomes as a template for PCR with internal primers. Our results indicate that the MROS3 gene is located in at least two copies tandemly arranged on the X chromosome with additional copy(ies) on the autosome(s), while MROS1, MROS2, and MROS4 are exclusively autosomal. The specificity of PCR products was checked by digestion with a restriction enzyme or reamplification using nested primers. Homology search of databases has shown the presence of five MROS3 homologues in A. thaliana, four of them arranged in two tandems, each consisting of two copies. We conclude that MROS3 is a low-copy gene family, connected with the proper pollen development, which is present not only in dioecious but also in other dicot plant species.

Amino Acid Sequence↗

PubMed1997

DNA extraction by zinc.

A fast, very simple and efficient method of DNA extraction is described which takes advantage of DNA sedimentation induced by millimolar concentrations of ZnCl2. The zinc-induced sedimentation is furthermore strongly promoted by submillimolar phosphate anion concentrations. Within <30 min, the method recovers >90% of DNA irrespective of whether a plasmid DNA or short oligonucleotides are the extracted material. The method works with plasmid DNA and oligonucleotide concentrations as low as 100 ng/ml and 10 microg/ml, respectively, without using any expensive facilities or toxic chemicals.

Buffers↗

PubMed1996

UV light-induced crosslinking of short DNA duplex strands: nucleotide sequence preferences and a prominent role of the duplex ends.

By various doses of UV light, we irradiated 32 short DNA duplexes having between 12 and 40 nucleotide residues in length, and monitored the induced crosslinks between the complementary DNA strands by denaturing polyacrylamide gel electrophoresis. The experiments revealed that the crosslinking was strongest with the alternating sequence of T and A and weaker with the alternating sequence of T and G (C and A in the complementary strand). On the other hand, GC blocks of any sequence provided undetectable amounts of interstrand crosslinks even at the highest doses of UV irradiation. The amount of crosslinked strands logarithmically increased with the UV dose but it did not depend on the oligonucleotide concentration, ionic strength, divalent magnesium or manganese cations and pH at least within the examined regions of the experimental conditions, unless the oligonucleotide denatured or isomerized into a unimolecular foldback. The extent of crosslinking also did not depend on the (dT-dA)n duplex length to indicate that the crosslink was predominantly localized at a specific duplex locus. Experiments with (dT-dA)8 "mutants" in which AT pairs were systematically replaced by GC pairs at various molecule positions, revealed that the crosslinking predominantly occurred at the oligo(dT-dA) duplex ends. The crosslinking is a direct method to detect duplexes of DNA, which is here, for example, demonstrated with the heteroduplex of (dT-dA)12 and (dT-dA)16.

Electrophoresis, Polyacrylamide Gel↗

PubMed1993

Tris buffer protects DNA backbone against breakage upon irradiation with ultraviolet light.

We show that Tris molecules protect DNA against nicking upon irradiation with ultraviolet light. However, the protective effect only concerns DNA backbone but not bases and it is observed in aqueous solution but not in formamide. Changes of pH or ionic strength due to Tris have no effect on the protection. The present observation has a practical importance for photofootprinting studies of DNA and its complexes with proteins but it can also serve as a basis for a development of a novel method reflecting DNA hydration and conformation.

Bacteriophage lambda↗