[Simple method of cloning eukaryote DNA: production of certain new ribosomal genes of Drosophila].
We propose a simple method which allows to receive a collection of clones containing recombinant plasmids. It is based on the ligation of the longer fragment of pBR332 formed by EcoRI and BamH1 with eukaryotic DNA (from Drosophila melanogaster embryo in this case) partially cleaved with EcoRI and BamHI. This approach gave us 10(4) colonies from 1 microgram of Drosophila DNA and 0.1 microgram of the BamHI--EcoRI "vector". About 0.5% of all clones carried the fragments of ribosomal genes with insertions in the 26S gene. Ribosomal genes lacking insertions did not enter the collection due to some peculiarities in their restriction map. The sites of cleavage are mapped in eight recombinant plasmide for HindIII, BamHI and EcoRI. These maps show that some insertions within 26S gene have not been cloned earlier. The mean length of cloned fragments is 11.8 kilobases, the mean number of EcoRI and BamHI restriction sites are 1.2 and 1.0, respectively. The electrophoretical screening of plasmids using cetyl trimethyl ammonium bromide was developed.
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant; D004331:Drosophila melanogaster / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D005822:Genetic Vectors; D010957:Plasmids; D012270:Ribosomes / Q000032:analysis*
D004274:DNA, Recombinant; D004247:DNA; D004262:DNA Restriction Enzymes
Prostoĭ metod klonirovaniia DNK éukariot: poluchenie nekotorykh novykh ribosomnykh genov drozofily.