In order to construct a physical map of the Hor1 locus of barley (Hordeum vulgare) high molecular weight DNA was prepared from leaf mesophyll protoplasts. Seventeen different restriction endonucleases containing CpG or CpXpG motifs in their recognition sequences were tested and ten proved useful for
Resolution ofAcanthamoeba castellaniichromosomes by pulsed field gel electrophoresis and construction of the initial linkage map
โ Scribed by David L. Rimm; Thomas D. Pollard; Philip Hieter
- Publisher
- Springer
- Year
- 1988
- Tongue
- English
- Weight
- 746 KB
- Volume
- 97
- Category
- Article
- ISSN
- 0009-5915
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โฆ Synopsis
Pulsed field gel electrophoresis has been used to resolve chromosome-sized DNA molecules in fungi and parasites but has not yet been used successfully to examine the chromosomes of other lower eukaryotes used extensively for biochemical research such as Acanthamoeba, Physarum, and Dictyostelium. Here we show an electrophoretic karyotype of the protozoan Acanthamoeba castellanii using orthogonal field alternating gel electrophoresis (OFAGE). There are about 20 small chromosomes ranging in size from 220 kb to greater than 2 Mb. We have assembled initial linkage groups assigning all of the cloned Acanthamoeba genes to chromosome-sized DNA molecules. Actin, suggested to have three or more non-allelic genes, maps to at least eight distinct chromosome bands. Two myosin II genes localize to two different chromosomal bands while myosin IB and 18S rRNA map to unresolved larger chromosomes.
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