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List of transformed lines
(fluorescent-tagged protamines)
(Belote & Pitnick Labs)Transformation vectors used:
pW8 = P-element vector with w+ (red eye) marker (Klemenz, et al., 1987)
pB{PUbnlsEGFP} = piggyBac vector with constitutive Ubiquitin-EGFP marker (Handler and Harrel, 2000)
pBac3xP3-EGFPaf = piggyBac vector with eye-speci c 3xP3-EGFP marker (Horn and Wimmer 2000)
D. melanogaster Protamine genes [(Jayaramaiah Raja and Renkawitz-Pohl (2005)]
ProtA = D. melanogaster Protamine A gene (also known as Mst(2)35Ba)
ProtB = D. melanogaster Protamine B gene (also known as Mst(2)35Bb)
If chromosome linkage is known, the chromosome is given at the end in parentheses (e.g., w ; P{w8, ProtA-DsRed-monomer, w+}1A(3) means the transgene is on chromosome 3)
I. D. melanogaster lines
Stocks in w1118 white-eye background:
Prot-RFP lines
- w ; P{w8, ProtA-DsRed-monomer, w+}1A(3)
- w ; P{w8, ProtB-DsRed-monomer, w+}16D(2)
- w ; P{w8, ProtB-DsRed-monomer, w+}50A(3)
Prot-GFP lines
- w ; P{w8, ProtA-EGFP, w+}19A(2)
- w ; P{w8, ProtA-EGFP, w+}19B(3)
- w ; P{w8, ProtB-EGFP, w+}75A(3)
- w ; P{w8, ProtB-EGFP, w+}27A(X)
- w ; P{w8, ProtB-EGFP, w+}27A(X); CyO / Bl
- w ; P{w8, ProtB-EGFP, w+}27A(X); TM3, Sb pp e / TM6B, Hu Tb e ca
Stocks in LHm wild-type background:
Prot-GFP lines
- LHm pBac{Ubnls-EGFP, ProtB-EGFP}6A (3)
- LHm pBac{Ubnls-EGFP, ProtB-EGFP}4A (3)
Prot-RFP line
- LHm BC-6 P{w8, ProtB-DsRed-monomer, w+}50A(3)
Stocks in CantonS wild-type background:
Prot-GFP lines
- CantonS pBac{3xP3-EGFP, ProtB-EGFP}27 (2)
- CantonS pBac{3xP3-EGFP, ProtB-EGFP}16B (3)
II. D. simulans lines
Prot-GFP line
Stock in wild-type background
- w+ ; pBac{3xP3-EGFP, ProtB-EGFP}11B
Prot-RFP line
Stock in w- background
- w ; P{w8, ProtB-DsRed-monomer, w+}3A
III. D. mauritiana lines Stocks in w- background
Prot-GFP lines
- w ; P{w8, ProtB-EGFP, w+}8A
- w ; P{w8, ProtB-EGFP, w+}22A
Prot-RFP line
- w ; P{w8, ProtB-DsRed-monomer, w+}13A
IV. Tribolium castaneum lines
(Droge-Young, E. M., J. M. Belote, G. S. Perez and S. Pitnick (2015) submitted)
All lines in peach (white eye) background;
Tribolium Protamine gene Tc2-4 = TcProt1 = GLEAN 07670 or LOC663849
TcProt1-GFP line
- p ; pBac{3xP3-EGFP, ProtTc2-4-EGFP}20
TcProt1-RFP line
- p ; pBac{3xP3-EGFP, ProtTc2-4-mCherry}4A / +
(This line is kept as heterozygous since homozygous transgenic chromosome is recessive lethal. Must be selected for presence of eye-GFP marker)
References:
Droge-Young, E. M., J. M. Belote, G. S. Perez and S. Pitnick. (2015) Mating system in uences on the mechanisms of postcopulatory sexual selection. Submitted.
Handler, A.M., Harrell, R.A., 2000. Transformation of the Caribbean fruit y with a piggyBac transposon vector marked with polyubiquitin-regulated GFP. Insect Biochem. Mol. Bioi. 31,199-205.
Horn, C., Wimmer, E.A., 2000. A versatile vector set for animal transgenesis. Devel. Genes Evol. 210, 630-637.
Jayaramaiah Raja, S., Renkawitz-Pohl, R., 2005. Replacement by Drosophila melanogaster protamines and Mst77F of histones during chromatin condensa- tion in late spermatids and role of Sesame in the removal of these proteins from the male pronucleus. Mol. Cell. Biol. 26, 6165–6177.
Klemenz, R., Weber, U., and Gehring, W.J. (1987). The white gene as a marker in a new P-element vector for gene transfer in Drosophila. Nucleic Acids Res. 15, 3947–3959.
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Download Resources:
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Belote Drosophila germline transformation protocol
DownloadConstruction of the fluorescently tagged Protamines A and B
DownloadConstruction of the tagged tribolium
DownloadTribolium Germline Transformation protocol
Download -
pB[3xP3EGFPaf] vector map
DownloadpB[PUbnlsEGFP] vector map
DownloadpW8 vector map
Download -
ProtamineA-GFP clone sequence
DownloadProtamineA-RFP clone sequence
DownloadProtamineB-GFP clone sequence
DownloadProtamineB-RFP clone sequence
DownloadTC Protamine1-GFP sequence
DownloadTC Protamine1-RFP clone sequence
Download
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