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D Ish-Horowicz

Publications and source records attributed to D Ish-Horowicz.

64 records · Page 4Linked to original sources
PubMed1981

Rapid and efficient cosmid cloning.

We present a procedure for cosmid cloning that allows rapid and efficient cloning of individual DNA fragments of between 32kb and 45kb. By appropriate treatment of the cloning vector, pJb8, we make left-hand and right-hand vector ends that are incapable of self-ligation but which accept dephosporylated insert DNA fragments. The inserted fragments are generated by partial digestion with MboI or Sau3A and are dephosphorylated to prevent ligation and insertion of non-contiguous fragments. The method eliminates the need to size the insert DNA fragments and prevents formation of clones containing short or multiple inserts. 1 microgram of target Drosophila DNA gives about 5 x 10(5) clones, with an average insert size of 38kb. We also describe a rapid and efficient method for preparing plasmid and cosmid DNA.

Animals↗
PubMed1981

Evolution of the 87A and 87C heat-shock loci in Drosophila.

Several species related to Drosophila melanogaster have two loci containing sequences which code for the major heat-shock protein of molecular weight 70,000 (hsp70). These sequences are arranged as two pairs of inverted repeats. The structure of the 87C locus in D. melanogaster seems to have arisen through DNA insertion. Hsp70-coding sequences at the two loci are not evolving independently. Gene conversion between hsp70 genes has apparently occurred both within and between loci.

Animals↗
PubMed1981

Genetic characterization of the region between 86F1,2 and 87B15 on chromosome 3 of Drosophila melanogaster.

The region between 86F1,2 and 87B15 on chromosome 3 of Drosophila melanogaster, which contains about 27 polytene chromosome bands including the 87A7 heat-shock locus, has been screened for EMS-induced visible and lethal mutations. We have recovered 268 lethal mutations that fall into 25 complementation groups. Cytogenetic localization of the complementation groups by deficiency mapping is consistent with the notion that each band encodes a single genetic function. We have also screened for mutations at the 87A7 heat shock locus, using a chromosome that has only one copy of the gene encoding the 70,000 dalton heat-shock protein (hsp70). No lethal or visible mutations at 87A7 were identified from 10,719 mutagenized chromosomes, and no female-sterile mutations at 87A7 were recovered from the 1,520 chromosomes whose progeny were tested for female fertility. We found no evidence that a functional hsp70 gene is required for development under laboratory conditions.

Animals↗
PubMed1979

Genetic and molecular analysis of the 87A7 and 87C1 heat-inducible loci of D. melanogaster.

Two different types of heat-inducible sequences are found at the cytogenetic loci 87A7 and 87C1 of D. melanogaster. One of these codes for the 70,000 dalton heat shock protein (hsp 70) and is found at both loci. The other type of sequence (alpha beta) codes for an RNA of unknown function and is found only at 87C1. We have completed a study of the organization of the two loci, using deficiencies that delete one or other locus, and have estimated the number of the hsp 70 genes at each locus. Thus in at least three strains of files there are a total of five coding sequences, three at 87C1 and two at 87A7. Restriction mapping of the coding regions at the two loci reveals that each of the two cytogenetic loci has its own characteristic coding sequence. The overall organization of the two loci appears to differ considerably. The alpha beta and hsp 70 heat-induced sequences at 87C1 are closely linked and are contained within two Eco RI restriction fragments.

Animals↗
PubMed1979

Deletion mapping of two D. melanogaster loci that code for the 70,000 dalton heat-induced protein.

Using deficiencies in D. melanogaster that lack either the 87A or 87C heat-induced puffs, we have shown that the 70,000 dalton heat-induced protein (hsp 70) is encoded at both these loci. Embryos deleted for one of the two loci retain the ability to make hsp 70 after heat shock, but deleting both loci eliminates synthesis of hsp 70. Thus both loci encode hsp 70 and can be active following heat shock. We have analyzed the proteins made by embryos lacking either 87A or 87C, and have compared the 87A- and 87C-coded hsp 70 by isoelectric focusing and tryptic peptide fingerprinting. The hsp 70 made by the two loci is very similar, although a variant tryptic peptide appears to be encoded only at 87C. Using deficiencies with slightly different breakpoints, we have mapped the 87A locus to band 87A7, the site of the 87A heat-induced puff. The 87C locus maps within 87C1.

Animals↗
PubMed1977

Deletions of two heat-activated loci in Drosophila melanogaster and their effects on heat-induced protein synthesis.

Deficiencies that delete two heat-induced puffs in Drosophila melanogaster have been isolated. Two deficiencies delete the puff arising from 87C1, and one deletes the two puffs at 87A and 87C1. Homozygotes for these deletions can be recognized by their abnormal, transparent malpighian tubules. The influence on heat-induced protein synthesis of deleting one or both of these puffs has been examined. Unexpectedly, deleting only the 87C1 puff has no apparent effect. Deleting both the 87A and 87C1 puffs eliminates synthesis of a 71,000 dalton protein. We map a coding locus for the 71,000 dalton heat-induced protein to 87A7-B3, a seven band region which also contains a heat-induced puff. The roles of the 87A and 87C1 puffs in coding for heat-induced proteins are discussed in the light of these results.

Animals↗