A set of P-element transformation vectors permitting the simplified generation of fluorescent fusion proteins inDrosophila melanogaster

Transcriptional regulation of the s38 chorion gene was studied using P element-mediated germline transformation. A 5·27 kb DNA fragment containing the s38 gene and 5′- and 3′-flanking sequences, was tested for its ability to be transcribed with correct developmental specificity. Five single-insert transformed lines were generated by microinjection of this DNA fragment cloned into a marked P element transformation vector. In each line, the transformed gene was transcribed according to the precise developmental pattern followed by the native s38 gene. The 1·3 kb at the 5′ end of this tested fragment was fused to the E. coli lac z gene. This fragment was also capable of initiating transcription of E. coli lac z RNA with the developmental profile of the native s38 gene. In vitro deletion studies are underway to determine which sequences in the 1·3 kb fragment are necessary for regulating the developmental expression of the gene.

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Correct Developmental and Tissue Specific Expression of a Cloned Alcohol Dehydrogenase Gene Introduced into the Drosophila Germ Line by P Element Transformation

Transfer ‖ Expression of Eukaryotic Genes ◽

10.1016/b978-0-12-284650-2.50009-2 ◽

1984 ◽

pp. 23-37

Author(s):

JAMES W. POSAKONY ◽

DAVID A. GOLDBERG ◽

TOM MANIATIS

Keyword(s):

Alcohol Dehydrogenase ◽

Germ Line ◽

P Element ◽

Specific Expression ◽

Tissue Specific ◽

Tissue Specific Expression ◽

Dehydrogenase Gene ◽

P Element Transformation

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P element transformation

The Dictionary of Genomics, Transcriptomics and Proteomics ◽

10.1002/9783527678679.dg09273 ◽

2015 ◽

pp. 1-1

Keyword(s):

P Element ◽

P Element Transformation

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Genetic and molecular characterization of a variegating hsp70-lacZ fusion gene in the euchromatic 31B region of Drosophila melanogaster

Genome ◽

10.1139/g01-038 ◽

2001 ◽

Vol 44 (4) ◽

pp. 698-707

Author(s):

Patrick Morcillo ◽

Ross J MacIntyre

Keyword(s):

Drosophila Melanogaster ◽

Heat Shock ◽

Fusion Gene ◽

Position Effect ◽

P Element ◽

Lacz Fusion ◽

Position Effect Variegation ◽

Salivary Gland Cells ◽

P Element Transformation

A hsp70lacZ fusion gene introduced into Drosophila melanogaster at the euchromatic 31B region by P-element transformation displayed a variegated expression with respect to the lacZ fusion protein in the salivary gland cells under heat-shock conditions. The variegation is also reflected by the chromosome puffing pattern. Subsequent transposition of the 31B P element to other euchromatic positions restored wild-type activity, that is, a nonvariegated phenotype. A lower developmental temperature reduced the amount of expression under heat-shock conditions, similar to genes undergoing position-effect variegation (PEV). However, other modifiers of PEV did not affect the expression pattern of the gene. These results show a novel euchromatic tissue-specific variegation that is not associated with classical heterochromatic PEV.Key words: Drosophila, euchromatic position effect, heat shock construct.

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The promoter of DNA puff gene II/9-1 of Sciara coprophila is inducible by ecdysone in late prepupal salivary glands of Drosophila melanogaster.

Cell Regulation ◽

10.1091/mbc.2.11.875 ◽

1991 ◽

Vol 2 (11) ◽

pp. 875-888 ◽

Cited By ~ 18

Author(s):

B Bienz-Tadmor ◽

H S Smith ◽

S A Gerbi

Keyword(s):

Drosophila Melanogaster ◽

Salivary Glands ◽

Dna Amplification ◽

P Element ◽

Control Of Gene Expression ◽

Evolutionarily Conserved ◽

Dna Puffs ◽

P Element Transformation ◽

Transgenic Drosophila

DNA puffs occur in Sciarid salivary gland chromosomes; they are sites of DNA amplification and intense transcription and they appear to encode secreted structural proteins needed for pupation. In this report we have used P-element transformation of Drosophila to study regulation of a Sciara DNA puff gene. We found that a 718-bp promoter fragment of DNA puff gene II/9-1 from Sciara coprophila directs expression of the bacterial reporter gene CAT in late prepupal salivary glands of transgenic Drosophila melanogaster. The identical tissue and analogous stage specificity indicate that some aspects of the ecdysone response are evolutionarily conserved between Drosophila and Sciara. When transgenic salivary glands are cultured in vitro, CAT activity is rapidly induced by ecdysone, suggesting direct control of gene expression by the ecdysone receptor. Putative stage-specific factors limit expression of the chimeric Sciara-CAT gene in transgenic Drosophila to late prepupae but not to third instar larvae when ecdysone titers are also high.

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New yeast/E. coli/Drosophila triple shuttle vectors for efficient generation of Drosophila P element transformation constructs

Gene ◽

10.1016/j.gene.2012.09.058 ◽

2012 ◽

Vol 511 (2) ◽

pp. 300-305 ◽

Cited By ~ 9

Author(s):

Achim Paululat ◽

Jürgen J. Heinisch

Keyword(s):

P Element ◽

Shuttle Vectors ◽

E Coli ◽

Efficient Generation ◽

New Yeast ◽

P Element Transformation

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Regulation of P-Element Transposase Activity in Drosophila melanogaster by hobo Transgenes That Contain KP Elements

Genetics ◽

10.1093/genetics/161.1.205 ◽

2002 ◽

Vol 161 (1) ◽

pp. 205-215 ◽

Cited By ~ 1

Author(s):

Michael J Simmons ◽

Kevin J Haley ◽

Craig D Grimes ◽

John D Raymond ◽

Joseph C L Fong

Keyword(s):

Drosophila Melanogaster ◽

P Element ◽

Drosophila Genome ◽

Maternal Transmission ◽

Alternate Splicing ◽

Hsp70 Promoter ◽

P Elements ◽

Naturally Occurring ◽

Element Activity ◽

Transformation Vectors

Abstract Fusions between the Drosophila hsp70 promoter and three different incomplete P elements, KP, SP, and BP1, were inserted into the Drosophila genome by means of hobo transformation vectors and the resulting transgenic stocks were tested for repression of P-element transposase activity. Only the H(hsp/KP) transgenes repressed transposase activity, and the degree of repression was comparable to that of a naturally occurring KP element. The KP transgenes repressed transposase activity both with and without heat-shock treatments. Both the KP element and H(hsp/KP) transgenes repressed the transposase activity encoded by the modified P element in the P(ry+, Δ2-3)99B transgene more effectively than that encoded by the complete P element in the H(hsp/CP)2 transgene even though the P(ry+, Δ2-3)99B transgene was the stronger transposase source. Repression of both transposase sources appeared to be due to a zygotic effect of the KP element or transgene. There was no evidence for repression by a strictly maternal effect; nor was there any evidence for enhancement of KP repression by the joint maternal transmission of H(hsp/KP) and H(hsp/CP) transgenes. These results are consistent with the idea that KP-mediated repression of P-element activity involves a KP-repressor polypeptide that is not maternally transmitted and that KP-mediated repression is not strengthened by the 66-kD repressor produced by complete P elements through alternate splicing of their RNA.

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