Inversions from the Bloomington Duplication Project
Kim Cook, Megan Deal, Jennifer Deal, Russ Garton, Rachel Andrade, Thom Kaufman & Kevin Cook
Bloomington Drosophila Stock Center
Indiana University
In this note, we will document the construction of a large series of chromosomal inversions on a C(1;Y) ("attached-XY") chromosome. We are using them in screens for Dp(1;Y) chromosomes, which we will describe later. We have used two slightly different approaches to generate the C(1;Y)N12, In(1)BSC chromosomes.
In the first approach, P{RS5} insertions distributed along the X chromosome were placed on C(1;Y)N12 by meiotic recombination to create a series of attached-XY chromosomes bearing single transposon insertions. P{RS3}CB-5805-3 was placed distal to the P{RS5} insertions by meiotic recombination to create a series of attached-XY chromosomes carrying two transposon insertions. By heat shocking females carrying one of these C(1;Y)N12, P{RS3}CB-5805-3 P{RS5} chromosomes and an autosomal P{70FLP} insertion, we could recover white-eyed progeny with a chromosome in which the P{RS5} and P{RS3} insertions had rearranged internally to delete FRT-flanked exons in the white gene markers. A second round of FLP-mediated recombination generated chromosomal inversions that reconstituted a white gene marker to give progeny with red eyes. All inversions were verified by polytene cytology.
The use of P{RS5} and P{RS3} to recover chromosomal aberrations was described in Golic & Golic, 1996 (FBrf0091061) and the isolation of the insertions we have used was described in Ryder et al., 2004 (FBrf0179424).
The following gives the name, distal insertion, proximal insertion, Release 5 coordinates and estimated cytological breakpoints of each inversion:
In(1)BSC24 P{RS3r}CB-5805-3 P{RS5r}5-HA-1961  X:387562 ;3266986 1B5;3D2
In(1)BSC3 P{RS3r}CB-5805-3 P{RS5r}5-SZ-3142  X:387562 ;3583172 1B5;3E3
In(1)BSC4 P{RS3r}CB-5805-3 P{RS5r}5-SZ-3655  X:387562 ;4825473 1B5;4D7
In(1)BSC5 P{RS3r}CB-5805-3 P{RS5r}5-SZ-4068  X:387562 ;5641035 1B5;5B6
In(1)BSC6 P{RS3r}CB-5805-3 P{RS5r}5-SZ-3429  X:387562 ;5882812 1B5;5D1
In(1)BSC8 P{RS3r}CB-5805-3 P{RS5r}5-HA-1616  X:387562 ;7090126 1B5;7A3
In(1)BSC9 P{RS3r}CB-5805-3 P{RS5r}5-SZ-3206  X:387562 ;8087225 1B5;7D18
In(1)BSC10 P{RS3r}CB-5805-3 P{RS5r}5-SZ-4103  X:387562 ;8924088 1B5;8C3
In(1)BSC25 P{RS3r}CB-5805-3 P{RS5r}5-HA-1967  X:387562 ;9580686 1B5;8F9
In(1)BSC11 P{RS3r}CB-5805-3 P{RS5r}5-SZ-4112  X:387562 ;10638967 1B5;9E1
In(1)BSC12 P{RS3r}CB-5805-3 P{RS5r}5-SZ-3419  X:387562 ;11347991 1B5;10B14
In(1)BSC13 P{RS3r}CB-5805-3 P{RS5r}5-SZ-4084  X:387562 ;11901120 1B5;11A1
In(1)BSC26 P{RS3r}CB-5805-3 P{RS5r}5-HA-1857  X:387562 ;12797208 1B5;11D1
In(1)BSC23 P{RS3r}CB-5805-3 P{RS5r}5-SZ-4109  X:387562 ;13536116 1B5;12A9
In(1)BSC14 P{RS3r}CB-5805-3 P{RS5r}5-SZ-4073  X:387562 ;14720102 1B5;12F4
In(1)BSC27 P{RS3r}CB-5805-3 P{RS5r}5-HA-1831  X:387562 ;15392986 1B5;13C5
In(1)BSC16 P{RS3r}CB-5805-3 P{RS5r}5-SZ-3670  X:387562 ;15985699 1B5;14A9
In(1)BSC17 P{RS3r}CB-5805-3 P{RS5r}5-HA-1737  X:387562 ;16549850 1B5;14F5
In(1)BSC19 P{RS3r}CB-5805-3 P{RS5r}5-HA-1561  X:387562 ;17576847 1B5;16C1
In(1)BSC20 P{RS3r}CB-5805-3 P{RS5r}5-HA-1134  X:387562 ;18400974 1B5;17C1
In(1)BSC21 P{RS3r}CB-5805-3 P{RS5r}5-SZ-3651  X:387562 ;19087625 1B5;18A7
In(1)BSC22 P{RS3r}CB-5805-3 P{RS5r}5-SZ-3656  X:387562 ;19781188 1B5;19A2
In(1)BSC28 P{RS3r}CB-5805-3 P{RS5r}5-HA-1907  X:387562 ;21961315 1B5;20C3
In the second approach, the proximal and distal insertions were first converted from P{RS3} or P{RS5} constructs with intact white gene markers to P{RS3r} or P{RS5r} constructs lacking intact white genes by heat shocking them in the presence of an autosomal P{hsFLP} insertion. Pairs of P{RS3r} and P{RS5r} insertions were placed in trans in females and these females were mated to males carrying an autosomal P{70FLP} insertion. Progeny were heat shocked as larvae and those carrying chromosomes with the P{RS5r} and P{RS3r} insertions placed in cis by meiotic recombination were recognized from their red eye clones resulting from somatic inversion events reconstituting a white gene marker. Because FLP recombinase also induced germ line recombination, we were able to recover inversion-bearing progeny as red-eyed flies. Once isolated and verified by polytene cytology, the inversions were placed on C(1;Y)N12 by meiotic recombination. This alternative method was much more efficient.
The following gives the name, distal insertion, proximal insertion, Release 5 coordinates and estimated cytological breakpoints of each inversion isolated by this second approach:
In(1)BSC1 P{RS3r}CB-5805-3 P{RS5r}5-HA-1598  X:387562 ;1837325 1B5;2B17
In(1)BSC2 P{RS3r}CB-5805-3 P{RS5r}5-SZ-3121  X:387562 ;2219975 1B5;2F6
In(1)BSC30 P{RS5r}5-SZ-4074 P{RS3r}CB-0332-3  X:417311 ;6589040 1B8;6C7
In(1)BSC32 P{RS3r}CB-5805-3 P{RS5r}5-HA-2919  X:387562 ;13178324 1B5;11E8
In(1)BSC33 P{RS3r}CB-5805-3 P{RS5r}5-HA-1883  X:387562 ;16686364 1B5;15A8
The following compiles all the inversions, their associated progenitor insertions, Release 5 breakpoints and predicted cytologies:
In(1)BSC24 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}5-HA-1961 FBti0029821 P{RS5r}5-HA-1961 FBti0032269  X:387562 ;3266986 1B5;3D2
In(1)BSC3 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}Mnt5-SZ-3142 FBti0029940 P{RS5r}5-SZ-3142 FBti0032388  X:387562 ;3583172 1B5;3E3
In(1)BSC4 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}CG40685-SZ-3655 FBti0030196 P{RS5r}5-SZ-3655 FBti0032644  X:387562 ;4825473 1B5;4D7
In(1)BSC5 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}CG31255-SZ-4068 FBti0030330 P{RS5r}5-SZ-4068 FBti0032778  X:387562 ;5641035 1B5;5B6
In(1)BSC6 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}5-SZ-3429 FBti0030070 P{RS5r}5-SZ-3429 FBti0032518  X:387562 ;5882812 1B5;5D1
In(1)BSC8 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}CG96505-HA-1616 FBti0029615 P{RS5r}5-HA-1616 FBti0032063  X:387562 ;7090126 1B5;7A3
In(1)BSC9 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}sdt5-SZ-3206 FBti0029969 P{RS5r}5-SZ-3206 FBti0032417  X:387562 ;8087225 1B5;7D18
In(1)BSC10 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}CG109625-SZ-4103 FBti0030354 P{RS5r}5-SZ-4103 FBti0032802  X:387562 ;8924088 1B5;8C3
In(1)BSC25 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}5-HA-1967 FBti0029826 P{RS5r}5-HA-1967 FBti0032274  X:387562 ;9580686 1B5;8F9
In(1)BSC11 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}ras5-SZ-4112 FBti0030360 P{RS5r}5-SZ-4112 FBti0032808  X:387562 ;10638967 1B5;9E1
In(1)BSC12 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}CG117275-SZ-3419 FBti0030063 P{RS5r}5-SZ-3419 FBti0032511  X:387562 ;11347991 1B5;10B14
In(1)BSC13 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}5-SZ-4084 FBti0030341 P{RS5r}5-SZ-4084 FBti0032789  X:387562 ;11901120 1B5;11A1
In(1)BSC26 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}CG44075-HA-1857 FBti0029758 P{RS5r}5-HA-1857 FBti0032206  X:387562 ;12797208 1B5;11D1
In(1)BSC23 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}NFAT5-SZ-4109 FBti0030358 P{RS5r}5-SZ-4109 FBti0032806  X:387562 ;13536116 1B5;12A9
In(1)BSC14 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}5-SZ-4073 FBti0030333 P{RS5r}5-SZ-4073 FBti0032781  X:387562 ;14720102 1B5;12F4
In(1)BSC27 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}Gmap5-HA-1831 FBti0029741 P{RS5r}5-HA-1831 FBti0032189  X:387562 ;15392986 1B5;13C5
In(1)BSC16 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}5-SZ-3670 FBti0030206 P{RS5r}5-SZ-3670 FBti0032654  X:387562 ;15985699 1B5;14A9
In(1)BSC17 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}r5-HA-1737 FBti0029693 P{RS5r}5-HA-1737 FBti0032141  X:387562 ;16549850 1B5;14F5
In(1)BSC19 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}CG325565-HA-1561 FBti0029572 P{RS5r}5-HA-1561 FBti0032020  X:387562 ;17576847 1B5;16C1
In(1)BSC20 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}CG64615-HA-1134 FBti0029420 P{RS5r}5-HA-1134 FBti0031869  X:387562 ;18400974 1B5;17C1
In(1)BSC21 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}CG141945-SZ-3651 FBti0030195 P{RS5r}5-SZ-3651 FBti0032643  X:387562 ;19087625 1B5;18A7
In(1)BSC22 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}amn5-SZ-3656 FBti0030197 P{RS5r}5-SZ-3656 FBti0032645  X:387562 ;19781188 1B5;19A2
In(1)BSC28 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}5-HA-1907 FBti0059582 P{RS5r}5-HA-1907 --  X:387562 ;21961315 1B5;20C3
In(1)BSC1 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}CG36005-HA-1598 FBti0029599 P{RS5r}5-HA-1598 FBti0032047  X:387562 ;1837325 1B5;2B17
In(1)BSC2 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}5-SZ-3121 FBti0029923 P{RS5r}5-SZ-3121 FBti0032371  X:387562 ;2219975 1B5;2F6
In(1)BSC30 P{RS5}arg5-SZ-4074 FBti0030334 P{RS5r}5-SZ-4074 FBti0032782 P{RS3}CB-0332-3 FBti0028059 P{RS3r}CB-0332-3 FBti0030509  X:417311 ;6589040 1B8;6C7
In(1)BSC32 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}HDAC45-HA-2919 FBti0059038 P{RS5r}5-HA-2919 --  X:387562 ;13178324 1B5;11E8
In(1)BSC33 P{RS3}CB-5805-3 FBti0028786 P{RS3r}CB-5805-3 FBti0031236 P{RS5}CG47685-HA-1883 FBti0029776 P{RS5r}5-HA-1883 FBti0032224  X:387562 ;16686364 1B5;15A8