Table 1.
Trojan exons, fly lines, and MiMIC insertions used or generated
| Transgene | Trojan Exon Plasmid | Genes/MiMIC Inserts for Fly Lines Made |
Developmentally Lethal2 |
Poor Viability3 |
|---|---|---|---|---|
| Gal4 | pBS-KS-attB2-SA(0)-T2A-Gal4-Hsp70 | RdlMI02620, ChaMI04508 | CCAP MI01341 (no recombinants) | |
| pBS-KS-attB2-SA(1)-T2A-Gal4-Hsp70 | GluRIIEMI01909, ShawMI01735, CCAPRMI05804, HR46MI04877, jebMI03124 | HR46MI00362, E75MI04895, abMI01579 | jebMI03124 | |
| pBS-KS-attB2-SA(2)-T2A-Gal4-Hsp70 | GluRIIBMI03631, dVGlutMI004979, Gad1MI09277 | Gad1MI09277 | ||
| pBS-KS-attB2-SA(0)-T2A-Gal41 | ||||
| pBS-KS-attB2-SA(1)-T2A-Gal41 | HR46MI04877, CCAPRMI05804 | E75MI04895, crcMI02300 | ||
| pBS-KS-attB2-SA(2)-T2A-Gal41 | dVGlutMI004979 | |||
| pC-(loxP2-attB2-SA(0)-T2A-Gal4-Hsp70) (i.e. “Singlet Donor”) | ||||
| pC-(loxP2-attB2-SA(1)-T2A-Gal4-Hsp70) (i.e. “Singlet Donor”) | ||||
| pC(loxP2-attB2-SA(2)-T2A-Gal4-Hsp70) (i.e. “Singlet Donor”) | ||||
| pC-(lox2-attB2-SA-T2A-Gal4-Hsp70)3 (i.e. “Triplet Donor”) | amonMI00899, RdlMI02957, Gycβ100B/STOPSMI01568 | CCAP MI01341 (no recombinants) | amonMI00899 | |
| pT-GEM(attP2-SA(0)-T2A-Gal4-Hsp70-lox2-3XP3-RFP) | pburs-Intron 1 | |||
| pT-GEM(attP2-SA(1)-T2A-Gal4-Hsp70-lox2-3XP3-RFP) | ||||
| pT-GEM(attP2-SA(2)-T2A-Gal4-Hsp70-lox2-3XP3-RFP) | ||||
| 3XGal80 | pBS-KS-attB2-SA(0)-T2A-3XGAL80-HSP70 | |||
| pBS-KS-attB2-SA(1)-T2A-3XGAL80-HSP70 | ||||
| pBS-KS-attB2-SA(2)-T2A-3XGAL80-HSP70 | dvGlutMI004979 | |||
| Gal4DBD | pBS-KS-attB2-SA(0)-T2A-GAL4DBD-Hsp70 | cacMI02836, ChaMI04508 | ||
| pBS-KS-attB2-SA(1)-T2A-GAL4DBD-Hsp70 | ||||
| pBS-KS-attB2-SA(2)-T2A-GAL4DBD-Hsp70 | dVGlutMI004979 | |||
| dVP16AD | pBS-KS-attB2-SA(0)-T2A-dVP16AD-Hsp70 | |||
| pBS-KS-attB2-SA(1)-T2A-dVP16AD-Hsp70 | ShawMI01735 | |||
| pBS-KS-attB2-SA(2)-T2A-dVP16AD-Hsp70 | ||||
| p65AD | pBS-KS-attB2-SA(0)-T2A-P65AD-Hsp70 | amonMI00899 | ||
| pBS-KS-attB2-SA(1)-T2A-P65AD-Hsp70 | ShawMI01735, RdlMI02957 | |||
| pBS-KS-attB2-SA(2)-T2A-P65AD-Hsp70 | Gad1MI09277 | |||
| QF2 | pBS-KS-attB2-SA(0)-T2A-QF2-Hsp70 | ChaMI04508 | amonMI00899 | |
| pBS-KS-attB2-SA(1)-T2A-QF2-Hsp70 | ||||
| pBS-KS-attB2-SA(2)-T2A-QF2-Hsp70 | dvGlutMI004979, Gad1MI09277 | |||
| LexA:QFAD | pBS-KS-attB2-SA(0)-T2A-LexA:QFAD-Hsp70 | ChaMI04508 | ||
| pBS-KS-attB2-SA(1)-T2A-LexA:QFAD-Hsp70 | ||||
| pBS-KS-attB2-SA(2)-T2A-LexA:QFAD-Hsp70 | dvGlutMI004979, Gad1MI09277 |
1
These Trojan Gal4 vectors lack the Hsp70 pA and are designed to retain a gene’s native transcription termination signals (e.g. when miRNA binding sites are present in the 3′ UTR). For two genes analyzed, we found no detectable differences in the expression patterns obtained with these constructs compared with those obtained with constructs containing the Hsp70 pA. We recommend use of the latter constructs to insure robust transcription termination unless mRNA downregulation is a known or suspected problem.
2
Lines for which recombinants died developmentally (or, as for CCAPMI01341, no recombinants were recovered)
3
Lines in which animals had impaired viability