Faculty Opinions recommendation of Editing Transgenic DNA Components by Inducible Gene Replacement in Drosophila melanogaster.

2016 ◽  
Author(s):  
Brian McCabe
Keyword(s):  
Drosophila Melanogaster ◽  
Gene Replacement ◽  
Dna Components ◽  
Inducible Gene

scholarly journals Editing Transgenic DNA Components by Inducible Gene Replacement inDrosophila melanogaster

Genetics ◽  
2016 ◽  
Vol 203 (4) ◽  
pp. 1613-1628 ◽  
Author(s):  
Chun-Chieh Lin ◽  
Christopher J. Potter
Keyword(s):  
Gene Replacement ◽  
Dna Components ◽  
Inducible Gene

The intron-containing hsp82 gene of the dimorphic pathogenic fungus Histoplasma capsulatum is properly spliced in severe heat shock conditions

1991 ◽  
Vol 11 (11) ◽  
pp. 5624-5630
Author(s):  
G Minchiotti ◽  
S Gargano ◽  
B Maresca
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Histoplasma Capsulatum ◽  
Pathogenic Fungus ◽  
Temperature Sensitive ◽  
General Phenomenon ◽  
High Degree ◽  
Beta Tubulin Gene ◽  
Inducible Gene

We have isolated and characterized a heat-inducible gene, hsp82, from the dimorphic pathogenic fungus Histoplasma capsulatum, which is a filamentous mold at 25 degrees C and a unicellular yeast at 37 degrees C. This gene, which has a high degree of homology with other members of the hsp82 gene family, is split into three exons and two introns of 122 and 86 nucleotides, respectively. Contrary to what has been demonstrated in Drosophila melanogaster, Saccharomyces cerevisiae, and other organisms, hsp82 mRNA in H. capsulatum is properly spliced during the severe heat conditions of 37 to 40 degrees C in the temperature-sensitive Downs strain. Splicing accuracy was also observed at 42 degrees C in the temperature-tolerant G222B strain, which showed no evidence of accumulation of primary transcripts. Furthermore, the intron containing the beta-tubulin gene is also properly spliced at the upper temperature range, suggesting that the lack of a block in splicing may be a general phenomenon in this organism.

scholarly journals P-element-induced interallelic gene conversion of insertions and deletions in Drosophila melanogaster.

1993 ◽  
Vol 13 (11) ◽  
pp. 7006-7018 ◽  
Author(s):  
D M Johnson-Schlitz ◽  
W R Engels
Keyword(s):  
Drosophila Melanogaster ◽  
Gene Replacement ◽  
P Element ◽  
Element Mobility ◽  
P Elements ◽  
Insertions And Deletions ◽  
Rapid Spread ◽  
White Locus ◽  
In Cis ◽  
P Transposon

We studied the process by which whd, a P-element insertion allele of the Drosophila melanogaster white locus, is replaced by its homolog in the presence of transposase. These events are interpreted as the result of double-strand gap repair following excision of the P transposon in whd. We used a series of alleles derived from whd through P-element mobility as templates for this repair. One group of alleles, referred to collectively as whd-F, carried fragments of the P element that had lost some of the sequences needed in cis for mobility. The other group, whd-D, had lost all of the P insert and had some of the flanking DNA from white deleted. The average replacement frequencies were 43% for whd-F alleles and 7% for the whd-D alleles. Some of the former were converted at frequencies exceeding 50%. Our data suggest that the high conversion frequencies for the whd-F templates can be attributed at least in part to an elevated efficiency of repair of unexpanded gaps that is possibly caused by the closer match between whd-F sequences and the unexpanded gap endpoints. In addition, we found that the gene substitutions were almost exclusively in the direction of whd being replaced by the whd-F or whd-D allele rather than the reverse. The template alleles were usually unaltered in the process. This asymmetry implies that the conversion process is unidirectional and that the P fragments are not good substrates for P-element transposase. Our results help elucidate a highly efficient double-strand gap repair mechanism in D. melanogaster that can also be used for gene replacement procedures involving insertions and deletions. They also help explain the rapid spread of P elements in populations.

scholarly journals Comparison of targeted-gene replacement frequencies in Drosophila melanogaster at the forked and white loci.

1996 ◽  
Vol 16 (7) ◽  
pp. 3535-3544 ◽  
Author(s):  
D H Lankenau ◽  
V G Corces ◽  
W R Engels
Keyword(s):  
Drosophila Melanogaster ◽  
Gene Conversion ◽  
Gene Targeting ◽  
Strand Break ◽  
Gene Replacement ◽  
P Element ◽  
White Gene ◽  
Success Rates ◽  
Conversion Frequency

P element-induced gene conversion has been previously used to modify the white gene of Drosophila melanogaster in a directed fashion. The applicability of this approach of gene targeting in Drosophila melanogaster, however, has not been analyzed quantitatively for other genes. We took advantage of the P element-induced forked allele, f(hd), which was used as a target, and we constructed a vector containing a modified forked fragment for converting f(hd). Conversion frequencies were analyzed for this locus as well as for an alternative white allele, w(eh812). Combination of both P element-induced mutant genes allowed the simultaneous analysis of conversion frequencies under identical genetic, developmental, and environmental conditions. This paper demonstrates that gene conversion through P element-induced gap repair can be applied with similar success rates at the forked locus and in the white gene. The average conversion frequency at forked was 0.29%, and that at white was 0.17%. These frequencies indicate that in vivo gene targeting in Drosophila melanogaster should be applicable for other genes in this species at manageable rates. We also confirmed the homolog dependence of reversions at the forked locus, indicating that P elements transpose via a cut-and-paste mechanism. In a different experiment, we attempted conversion with a modified forked allele containing the su(Hw) binding site. Despite an increased sample size, there were no conversion events with this template. One interpretation (under investigation) is that the binding of the su(Hw) product prevents double-strand break repair.

scholarly journals The intron-containing hsp82 gene of the dimorphic pathogenic fungus Histoplasma capsulatum is properly spliced in severe heat shock conditions.

1991 ◽  
Vol 11 (11) ◽  
pp. 5624-5630 ◽  
Author(s):  
G Minchiotti ◽  
S Gargano ◽  
B Maresca
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Histoplasma Capsulatum ◽  
Pathogenic Fungus ◽  
Temperature Sensitive ◽  
General Phenomenon ◽  
High Degree ◽  
Beta Tubulin Gene ◽  
Inducible Gene

We have isolated and characterized a heat-inducible gene, hsp82, from the dimorphic pathogenic fungus Histoplasma capsulatum, which is a filamentous mold at 25 degrees C and a unicellular yeast at 37 degrees C. This gene, which has a high degree of homology with other members of the hsp82 gene family, is split into three exons and two introns of 122 and 86 nucleotides, respectively. Contrary to what has been demonstrated in Drosophila melanogaster, Saccharomyces cerevisiae, and other organisms, hsp82 mRNA in H. capsulatum is properly spliced during the severe heat conditions of 37 to 40 degrees C in the temperature-sensitive Downs strain. Splicing accuracy was also observed at 42 degrees C in the temperature-tolerant G222B strain, which showed no evidence of accumulation of primary transcripts. Furthermore, the intron containing the beta-tubulin gene is also properly spliced at the upper temperature range, suggesting that the lack of a block in splicing may be a general phenomenon in this organism.

scholarly journals A POSSIBLE ROLE OF REPETITIOUS DNA IN RECOMBINATORY JOINING DURING CHROMOSOME REARRANGEMENT IN DROSOPHILA MELANOGASTER

Genetics ◽  
1975 ◽  
Vol 79 (3) ◽  
pp. 467-470
Author(s):  
C S Lee
Keyword(s):  
Drosophila Melanogaster ◽  
In Situ Hybridization ◽  
Strong Correlation ◽  
Chromosome Rearrangement ◽  
Chromosome Breaks ◽  
Recombination Processes ◽  
X Irradiation ◽  
Dna Components

ABSTRACT It is postulated that certain repetitious DNA components play a role in the recombination processes during chromosome rearrangements. When the distribution of silver grain densities after the in situ hybridization of repetitious DNA (Rudkin and Tartof 1973) and the distribution of chromosome breaks due to X-irradiation (Kaufmann 1946) are compared, a strong correlation is found for the euchromatic portion of the D. melanogaster salivary X chromosome. These observations justify the postulate above that certain repetitious DNA provides homologous regions in the DNA of broken chromosome ends necessary for proper recombinatory joining.

Glucocorticoid-inducible gene expression vectors for use in Drosophila melanogaster Schneider 2 cells

2004 ◽  
Vol 13 (2) ◽  
pp. 205-211 ◽  
Author(s):  
J. Poels ◽  
A. Martinez ◽  
M.-M. Suner ◽  
A. De Loof ◽  
S. J. Dunbar ◽  
...  
Keyword(s):  
Gene Expression ◽  
Drosophila Melanogaster ◽  
Expression Vectors ◽  
Inducible Gene Expression ◽  
Inducible Gene

scholarly journals Complex interactions among members of an essential subfamily of hsp70 genes in Saccharomyces cerevisiae.

1987 ◽  
Vol 7 (7) ◽  
pp. 2568-2577 ◽  
Author(s):  
M Werner-Washburne ◽  
D E Stone ◽  
E A Craig
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Drosophila Melanogaster ◽  
Large Family ◽  
Temperature Sensitive ◽  
Gene Products ◽  
Wild Type ◽  
Functional Protein ◽  
Complex Interactions ◽  
Intact Copy ◽  
Inducible Gene

Saccharomyces cerevisiae contains a large family of genes related to hsp70, the major heat shock-inducible gene of Drosophila melanogaster. One subfamily, identified by sequence homology, contains four genes, SSA1, SSA2, SSA3, and SSA4 (formerly YG100, YG102, YG106, and YG107, respectively). Previous studies showed that strains containing mutations in SSA1 and SSA2 are temperature sensitive for growth. SSA4, which is normally heat inducible and not expressed during vegetative growth, is expressed at high levels in ssa1 ssa2 strains at 23 degrees C. We constructed mutations in SSA3 and SSA4 and analyzed strains carrying mutations in the four genes. Strains carrying mutations in SSA3 SSA4 or SSA3 and SSA4 were indistinguishable from the wild type. However, ssa1 ssa2 ssa4 strains were inviable. SSA3, like SSA4, is a heat-inducible gene that is not normally expressed at 23 degrees C. Nevertheless, an intact copy of SSA3 regulated by the constitutive SSA2 promoter was capable of rescuing a ssa1 ssa2 ssa4 strain. This indicates that SSA3 encodes a functional protein and that the SSA1, SSA2, SSA3, and SSA4 gene products are functionally similar.

Complex interactions among members of an essential subfamily of hsp70 genes in Saccharomyces cerevisiae

1987 ◽  
Vol 7 (7) ◽  
pp. 2568-2577 ◽  
Author(s):  
M Werner-Washburne ◽  
D E Stone ◽  
E A Craig
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Drosophila Melanogaster ◽  
Large Family ◽  
Temperature Sensitive ◽  
Gene Products ◽  
Wild Type ◽  
Functional Protein ◽  
Complex Interactions ◽  
Intact Copy ◽  
Inducible Gene

Saccharomyces cerevisiae contains a large family of genes related to hsp70, the major heat shock-inducible gene of Drosophila melanogaster. One subfamily, identified by sequence homology, contains four genes, SSA1, SSA2, SSA3, and SSA4 (formerly YG100, YG102, YG106, and YG107, respectively). Previous studies showed that strains containing mutations in SSA1 and SSA2 are temperature sensitive for growth. SSA4, which is normally heat inducible and not expressed during vegetative growth, is expressed at high levels in ssa1 ssa2 strains at 23 degrees C. We constructed mutations in SSA3 and SSA4 and analyzed strains carrying mutations in the four genes. Strains carrying mutations in SSA3 SSA4 or SSA3 and SSA4 were indistinguishable from the wild type. However, ssa1 ssa2 ssa4 strains were inviable. SSA3, like SSA4, is a heat-inducible gene that is not normally expressed at 23 degrees C. Nevertheless, an intact copy of SSA3 regulated by the constitutive SSA2 promoter was capable of rescuing a ssa1 ssa2 ssa4 strain. This indicates that SSA3 encodes a functional protein and that the SSA1, SSA2, SSA3, and SSA4 gene products are functionally similar.

P-element-induced interallelic gene conversion of insertions and deletions in Drosophila melanogaster

1993 ◽  
Vol 13 (11) ◽  
pp. 7006-7018
Author(s):  
D M Johnson-Schlitz ◽  
W R Engels
Keyword(s):  
Drosophila Melanogaster ◽  
Gene Replacement ◽  
P Element ◽  
Element Mobility ◽  
P Elements ◽  
Insertions And Deletions ◽  
Rapid Spread ◽  
White Locus ◽  
In Cis ◽  
P Transposon

We studied the process by which whd, a P-element insertion allele of the Drosophila melanogaster white locus, is replaced by its homolog in the presence of transposase. These events are interpreted as the result of double-strand gap repair following excision of the P transposon in whd. We used a series of alleles derived from whd through P-element mobility as templates for this repair. One group of alleles, referred to collectively as whd-F, carried fragments of the P element that had lost some of the sequences needed in cis for mobility. The other group, whd-D, had lost all of the P insert and had some of the flanking DNA from white deleted. The average replacement frequencies were 43% for whd-F alleles and 7% for the whd-D alleles. Some of the former were converted at frequencies exceeding 50%. Our data suggest that the high conversion frequencies for the whd-F templates can be attributed at least in part to an elevated efficiency of repair of unexpanded gaps that is possibly caused by the closer match between whd-F sequences and the unexpanded gap endpoints. In addition, we found that the gene substitutions were almost exclusively in the direction of whd being replaced by the whd-F or whd-D allele rather than the reverse. The template alleles were usually unaltered in the process. This asymmetry implies that the conversion process is unidirectional and that the P fragments are not good substrates for P-element transposase. Our results help elucidate a highly efficient double-strand gap repair mechanism in D. melanogaster that can also be used for gene replacement procedures involving insertions and deletions. They also help explain the rapid spread of P elements in populations.

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