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Reference
Citation	Ring, B., Garza, D. (2003.6.10). PBac Constructs from the Project 'Genome Wide Dispersal of Half-P Elements in Drosophila Using PiggyBac.
FlyBase ID	FBrf0159884
Type of publication	Personal communication to FlyBase
Offprint Available	No
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Text of personal communication	From matthewk@XXXXXXXXXXXXXXX Tue Jun 10 20:14:48 2003 Subject: PBac Half-P Constructs To: flybase-updates@XXXXXXXXXXXXXXX Cc: brian.ring@XXXXXXXXXXXXXXX Personal communication from: Brian Ring and Dan Garza, Florida State University, Novartis To: Bloomington Drosophila Stock Center Subject: PBac Constructs from the Project 'Genome Wide Dispersal of Half-P Elements in Drosophila Using PiggyBac' Dated: 23 May 2003 Background: This is the information needed to curate the two piggyBac constructs that are present in the set of PBac insertion lines donated to the Bloomington Stock Center by Brian Ring and Dan Garza. The primary reference is Brian's dissertation, FBrf0152434 (Chapter 4): Ring, Brian C., 'Functional Analysis of the Drosophila Genome Using Transposable Elements'. Florida State University. 2001 pgs. 130-199. Data on the insertion lines themselves will be submitted separately as a computer file. Information communicated: 1) PBac{5HPw⁺} This is a 5' half-P construct marked with w^+mC. The minimal 5' P-element cis acting sequences and 8 bp target site duplication derived from P{lacW} were cloned into the BglII site of Al Handler's pB{Dmw} construct (Handler and Harrell, 1999, Insect Molec. Biol. 8:449--457; FBrf0123037) and transformed into flies. Handler created pB{Dmw} by cloning the w^+mC allele (4.2-kb EcoRI fragment) into the HpaI site of the original piggyBac plasmid p3E1.2 (Cary et al., 1989, Transposon mutagenesis of baculoviruses: analysis of Trichoplusiani transposon IFP2 insertions within the FP-Locus of nuclear polyhedrosis viruses. Virology 161: 8--17). The white mini-gene interrupts the piggyBac transposase open reading frame, but otherwise leaves the piggyBac element intact, with the respective promoters in opposite orientation (per Handler and Harrell, 1999). Insertions of this construct used for the screens: PBac{5HPw⁺}X7 \- This homozygous viable and fertile X chromosome insertion was the progenitor of the A series of insertions. PBac{5HPw⁺}X10 \- This homozygous viable and fertile X chromosome insertion was the progenitor of the B series of insertions. 2) PBac{3HPy⁺} This is a 3' half-P construct marked with y^+mDint. The minimal 3' P-element cis acting sequences and 8 bp target site duplication derived from P{lacW} were cloned into the 3' end of the yellow mini-gene (Geyer and Corces, 1987, Genes Dev. 1: 996--1004; FBrf0046293) in plasmid yellow-BSX, a generous gift from Pam Geyer, to form the intermediate plasmid p3HPy⁺. This DNA was then shuttled into a modified site in p3E1.2 (Cary et al., 1989, see above) to form pPBac{3HPy⁺}, which was transformed into flies. The resulting yellow mini-gene and 3' P-element interrupt the piggyBac gene and reside in opposite orientation with respect to promoters. Because intronic sequences necessary for y⁺ expression in bristles (all types), tarsal claw and arista are not present in y^+mDint, y^- flies transformed with PBac{3HPy⁺} lack y⁺ pigmentation in these structures. Insertions of this construct used for the screens: PBac{3HPy⁺}X0 and PBac{3HPy⁺}X3 \- These homozygous viable and fertile X chromosome insertion were the progenitors of the C series of insertions.
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Language of publication	English
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Language of publication	English
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Data from Reference
Alleles (2)
	w^+mC y^+mDint2
Constructs (2)
	PBac{3HPy⁺} PBac{5HPw⁺}
Genes (3)
	Tni\piggyBac\T w y
Insertions (349)
	All Insertions relating to this reference
Natural transposons (1)
	Tni\piggyBac