ABSTRACT
In an effort to generate a genome-wide set of high-quality polymorphic markers for the rat, we used the marker-selection method, which has already been proven useful for the development of markers, especially for the human genome. Small-insert (300-900 bp) rat genomic libraries were constructed with an estimated complexity of three genome equivalents and enriched for short tandem repeat sequences (STRs). The enriched libraries were found to contain 45% (CA)n and 27% (GATA)n, representing at least a 50-fold enrichment over unselected small insert genomic libraries. A subset of 2160 STR-containing clones, primarily of the (GATA)n class of repeats, were sequenced. PCR primers flanking the repeats were synthesized from some of the sequences from the (CA)n and (GATA)n classes of STRs and tested for polymorphism in a panel of eight inbred rat strains. This strategy yielded 147 polymorphic markers, which mapped with high odds to all chromosomes by linkage in three F2 populations. The integration of these STR markers with other rat genetic markers and mapping reagents will facilitate the mapping of disease genes in the rat and the identification of loci associated with complex mammalian phenotypes.
Subject(s)
Genetic Markers/genetics , Genome , Genomic Library , Tandem Repeat Sequences/genetics , Animals , Base Sequence , Chromosome Mapping , Chromosomes/genetics , Cloning, Molecular , DNA/chemistry , DNA/genetics , Dinucleotide Repeats/genetics , Microsatellite Repeats/genetics , Molecular Sequence Data , Polymorphism, Genetic , Rats , Rats, Inbred BN , Rats, Inbred Lew , Rats, Inbred SHR , Rats, Inbred WKY , Rats, Sprague-DawleyABSTRACT
The activity of 10 enzymes separated by acrylamide disc gel electrophoresis of leaf and stem extracts from Dianthus grown under summer and winter conditions was studied. While banding was constant and highly reproducible under each environment, differences between the 3 cultivars and between the tissues were evident. No significant differences in the isozyme patterns of glutamate dehydrogenase, 6-phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase, malate dehydrogenase, and catalase were observed between the 2 environments. Loss of activity was observed under winter conditions with amylase and lactate dehydrogenase and loss of certain isozymic components was evident with acid phosphatase and esterase. Prominent changes were observed in peroxidase isozymes, the hardy cultivars developing additional isozymic components under winter conditions. Only minor changes in the total protein banding were seen. The enzymes showed considerable stability in those tissues killed by the freezing conditions.
ABSTRACT
Simplified but highly reproducible extraction and electrophoretic procedures have been developed for plant stem and leaf proteins using recent chemical and technical advances. The method is applied to the separation of the basic, water-soluble proteins found in stem and leaf tissues of 3 Dianthus clones. While most of the highly reproducible protein bands appear in all 3 selections, and many are also common to both leaf and stem tissue, others are characteristic for the variety or tissue.
