In situ hybridization of cRNA transcribed from cloned D. melanogaster heat shock sequences to D. hydei chromosomes has shown that the D. hydei locus 2--32 A corresponds to the D. melanogaster locus 87 A/C and the D. hydei locus 2--36 A to the D. melanogaster locus 95 D, while the D. hydei locus 4--81 B corresponds to the D. melanogaster locus 63 BC. No hybridization to D. hydei chromosomes was found with cRNA transcribed from a clone containing the alpha beta sequences encoded by the D. melanogaster locus 87 C. Neither D. melanogaster heat shock RNA nor D virilis heat shock RNA hybridized significantly to the D. hydei heat shock locus 2--48 B. Furthermore, D. hydei heat shock RNA did not hybridize to the cytological homologs of locus 2--48 B found in D. repleta or in D. virilis. D. hydei heat shock. RNA did hybridize to the cytological homologs of locus 2--48 B in D. neohydei and D. eohydei, both of which belong to the hydei subgroup.
We have previously isolated a 500 bp-long cDNA clone, NO9-15, which is derived from a nuclear transcript originating from the heat shock locus 2-48B of Drosophila hydei (Peters et al. 1982). Sequence analysis shows that this clone carries 4 complete copies and 1 partial copy of a 115 bp repeat unit. The repeats are closely homologous with a maximal sequence divergence of about 10%. The sequence does not contain an open reading frame. The genomic organization of heat shock locus 2-48B, as probed with the cloned cDNA sequence NO9-15, is highly polymorphic. Four different allelic arrangements have been found in different inbred strains. A number of genomic clones isolated from region 2-48B, both in phage lambda and in cosmid vectors, all differ in length, mainly due to varying numbers of the NO9-15 repeat unit. These differences are found primarily in the proximal region of the locus. The transcribed region of these clones includes the distal sequence flanking the NO9-15 repeat as well as the NO9-15 repeat itself. An oligo A stretch was found between the distal flanking sequence and the NO9-15 repeat region.
cDNA, copied from nuclear RNA isolated from heat shocked Drosophila hydei cells, has been cloned. From this collection of clones a clone, N09-15, with a 450 bp insert has been isolated that hybridizes in situ to the heat shock locus-2-48B of Drosophila hydei. The N09-15 sequence is present in two different genomic arrangements, as shown by restriction mapping, in our wild type D. hydei population. These genomic arrangements are allelic. Both alleles contain multiple copies of the N09-15 sequence but differ in their lengths and in the distribution of Msp I and Taq I sites.
Kits of five different suppliers, composed according to the Dutch recommendations for determination of the enzyme activity of alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, gamma-glutamyltransferase, alkaline phosphatase, and creatine kinase, were intercompared. Activity concentrations of the enzymes in human sera were measured under defined conditions, evaluated, and related to the actual composition of the kits. Concentrations of all kit components were determined by various analytical techniques. The overall results of the activity measurements and the composition of at least three kits inter-agree well. We found deviations as great as 10% in our analytical evaluation of the kits of the other two suppliers, which can partly be accounted for.
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