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G M Guild

Publications and source records attributed to G M Guild.

28 records · Page 2Linked to original sources
PubMed1986

Molecular analysis of the ecdysterone-inducible 2B5 "early' puff in Drosophila melanogaster.

The 2B5 region of the X-chromosome in Drosophila melanogaster plays a developmentally important role in the ecdysterone-triggered response of the late third instar salivary gland. Using a combination of transposon-tagging and chromosomal walking techniques, we have isolated 231 kb of contiguous genomic DNA sequences corresponding to this region. We have more precisely aligned this DNA to the 2B1,2 to 2B5-6 interval of the cytogenetic map by locating the position of three well-characterized chromosomal breakpoints by in situ hybridization and genomic DNA blotting experiments. Labeled cDNA, synthesized from poly(A)+ RNA isolated from hormone-induced salivary gland and imaginal disc tissues and hybridized to the cloned DNA, demonstrated that the ecdysterone-inducible sequences mapped to DNA segments corresponding to the 2B3,4 to 2B5-6 interval. Although some of these sequences were inducible in only one tissue type, many were found to be inducible in both salivary glands and imaginal discs. RNA blotting experiments have detected a major 4.5-kb RNA which is hormone inducible in the larval salivary gland and whose quantitative induction is not inhibited by cycloheximide. Thus, the 4.5-kb RNA represents at least one product from the ecdysterone-responsive 2B5 "early' puff.

Animals↗
PubMed1986

Poly(A) shortening of coregulated transcripts in Drosophila.

The 71E ecdysterone-regulated puff of Drosophila melanogaster contains a cluster of six coregulated "late" genes which are expressed in the prepupal salivary gland. The resulting transcripts exhibit a decrease in their length during the 12-hr period in which they accumulate. Using the enzyme ribonuclease H, we show that this size decrease is a result of a progressively shorter poly(A) tract and suggest that these transcripts undergo an active sequential shortening of their poly(A) tracts in prepupal salivary glands. It is interesting to note that this shortening precedes the complete loss of these transcripts from the RNA population.

Animals↗
PubMed1984

Larval salivary gland secretion proteins in Drosophila. Identification and characterization of the Sgs-5 structural gene.

The 90BC locus on the polytene chromosomal map of Drosophila melanogaster contains the structural gene for a third-instar, salivary gland-specific, polyadenylated RNA (the group V RNA). This also belongs to the intermolt puff set whose dispersed and co-ordinately regulated members are (1) transcriptionally active in the salivary gland during the third-instar developmental stage and (2) comprise (at least in part) the structural genes for a set of salivary gland secretion proteins. Previous developmental studies of the group V intermolt gene (located cytogenetically within the 90B3-8 interval) suggest that it controls the expression of a salivary gland secretion protein. By analyzing different D. melanogaster laboratory stocks for variation in group V gene expression, we have been able to correlate the presence of the group V RNA with the salivary gland secretion protein P4. In vitro translation experiments show that the salivary gland messenger RNA population derived from a stock that fails to synthesize the group V RNA does not direct the synthesis of a polypeptide similar in molecular weight to protein P4. In addition, cloned genomic DNA segments complementary to the group V RNA are capable of arresting the in vitro translation of this protein. Comparative two-dimensional fractionation of cysteine-labeled, protease-generated peptides shows that (1) the in vitro translation product arrested by group V gene DNA is biochemically very similar to or identical with the salivary gland secretion protein P4, and (2) protein P4 is equivalent to the salivary gland secretion protein previously designated SGS-5. Since designations of the latter type have been employed in naming the genetic loci that represent the structural genes for the salivary gland secretion protein gene set, the group V gene (previous designation) represents the SGS-5 structural gene and its appropriate genetic designation should now be Sgs-5.

Animals↗
PubMed1984

Molecular analysis of a developmentally regulated gene which is expressed in the larval salivary gland of Drosophila.

The polytene genome of the larval salivary gland of Drosophila undergoes dramatic alterations in localized transcriptional activity late in third-instar development. One of these changes involves the 20-OH ecdysone-mediated and coordinate repression of a dispersed set of intermolt puff sites. The DNA from one of these loci (that located within the 90BC interval) has been isolated by molecular cloning techniques. This DNA was shown to be present one time per haploid Drosophila genome and to contain a gene (designated the Group V gene) which is developmentally regulated. The Group V transcript was shown to accumulate only during third-instar larval development and to be located exclusively in the salivary gland. These results are consistent with the hypothesis that the Group V gene codes for a salivary gland glue protein.

Animals↗
PubMed1980

A new high-capacity cosmid vector and its use.

The construction of a cosmid, MUA-3, designed for the convenient cloning of eukaryotic DNA segments up to 48 kb in length is described. The cosmid contains all of the plasmid pBR322 with approx. 400 bases of lambda DNA, including the cohesive end site, inserted at the pBR322 PstI endonuclease recognition site. Methods for using this vector to construct several types of Drosophila melanogaster genomic DNA libraries are given, and libraries made by these methods are characterized. A sheared Drosophila DNA-EcoRI linker library is shown to stably maintain average Drosophila DNA inserts of over 40 kb and up to 48 kb, and the efficiency of producing clones by a partial restriction and ligation method is shown to be over 3 X 10(5) clones/microgram of Drosophila DNA.

Bacteriophage lambda↗
PubMed1977

The significance and nature of defective interfering viruses.

Deletions in viral genomes appear to be a common occurrence in the replication of all DNA and RNA viruses which have been adequately studied. Such defective genomes can replicate in the presence in the same cell of a helper virus as long as the deletion does not involve the initiation site for genome replication. Coinfection of a cell with defective and "normal" infectious virus leads to reduction in the yield of the latter. The nature of DI viruses and genomes found in Sindbis virus-infected vertebrate cells during "undiluted passage" series is discussed. This procedure leads to the accumulation of progressively shorter viral RNA genomes with internal deletions. The enrichment is limited to genome lengths which are integral fractions (1/2, 1/3, 1/4, etc.) of the complete genome, and these are also found in viral particles released at the corresponding passage levels. It is believed that the selective accumulation of these fragments is governed by constraints of assembly which demand that one full genome equivalent be packaged in a released particle. In contrast to vertebrate cells, cultured mosquito cells do not seem to produce or "recognize" DI particles. Possible implications for the epidemiology of arthropod-transmitted alphaviruses are presented.

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