1. Lab Meeting
Bo Shi

2. Outlines 1 Dpr and DIP primer testing 2 Confocus imaging
3 Single cell dissociation
4 Transgenetic flies (just starting)
5 Dprs and DIPs recombining flies (just starting)

3. 1 Dpr and DIP primer testing
The primers of 12 Dprs have been tested, and can be used for the next step. (The other 8 or 20 Dprs has been tested before my last lab meeting.)
10 of 11 DIPs also can be used. (exception: CG40378)

4. 2nd round PCR images No. 1 2 3 4 5 6 Genes Dpr1 Dpr 3 Dpr 7 Dpr 8
Dpr 10-new
cDNA
201
248
270
220
457
225
gDNA
N/A
8661
5056
888

5. No. 1 2 3 4 5 6
Genes
Dpr11-new
Dpr15-new
Dpr16-new
Dpr17-new
Dpr 19
Dpr 20
cDNA
242
208
370
273 X
gDNA
826
3500
801
2124

6. No. 1 2 3 4 5 6
Genes
Dpr19-new
Dpr 20-new
DIP-α(1)
DIP-γ(3)
DIP-δ(4)
DIP-ε(5)
cDNA
202
337
288
385
318
311
gDNA
N/A
1332
841
1732
5272

7. No. 1 2 3 4 5 6 Genes DIP-ζ(6) DIP-η(7) DIP-θ(8) DIP-ι(9) cDNA 274 X
CG31814
CG40378
cDNA
274 X
320
253
gDNA
N/A
3225

8. No. 1 2 3 Genes DIP-β(2) cDNA 309 /224 324 X gDNA 15049 N/A 1 2 3
DIP-η(7)-new
CG40378-new
cDNA
309 /224
324 X
gDNA
15049
N/A

9. 2 Confocus imaging ort-c1a-lv Tm20: YW ; 17D01-Flp ; lot-stop-mCD8-GFP+ ;
♂ 122 ;
C1-Gal4, UAS-mCD8-Cherry +
FRT82B, Gal80
FRT82B, Tsc1
Tsc mutation, Tm20

10. 3 Single cell dissociation
FH1
♀ YW ;
17D01-Flp
Cyo ; +
Tm2 or Tm6B X
♂ YW ;
ort-c1a-lv
lot-stop-mCD8-GFP
ort-c1a-lv
F1: YW ;
17D01-Flp ;
lot-stop-mCD8-GFP +

11. 2nd round PCR images mCD8 1 2 3 4 5 6 7 8 vGlut 1 2 3 4 5 6 7 8
Primers
cDNA
gDNA
mCD8
266
vGlut
339
807

12. 4 Transgenetic flies Rh3-syb-spGFP1-10 Rh4-syb-spGFP1-10
UAS-YFP-sp1-10cyto

13. 5 Dprs and DIPs recombining flies
57B1-57B2
FRT42
Dpr20 3L
61D2-61D2
FRT2A
Dpr3 2L
22C1-22C3
FRT40
CG14010
25F1-25F1
CG31708
29F8-30A1

14. Future Work 1 Complete CG40378 primers design and test.
2 Test DIPs and Dprs expression in the medulla.
3 Test DIPs and Dprs expression in Tm20, Dm8 & Tm5C.
4 Inditify which Ach receptor expresses in Tm20.
5 Screen some adhesion molecule mutations in Tm20.

15. 6 Continue to pick Transgenetic flies
7 Continue to recombine Dprs and DIPs flies with FRT flies
8 Obtain the specificity of Tm20 in the line of
ort-c1a-lv
F1: YW ;
17D01-Flp ;
lot-stop-mCD8-GFP +

17. Transgenetic flies X YW
F1: 1♂ red eye fly X
YW; ;
Cyo
Tm6B
Tm2 B1 3♀
If transgne is on the ChX, all females are red eyes and males are white eyes in the F2 geneation.
If Transgene is on the on the Ch2, Ch3 or Ch4, then:
F2: 1♂ red eye fly X
YW ; ;
Cyo
Tm6B
Tm2 B1 3♀

18. 3 Single cell dissociation
17Dol-Flp +
ort-cla-lv
lot-stop-mCD8-GFP
♀ YW; ; X
♂ YW; ;
Cyo
Tm2 or Tm6B
Cyo
Tm2 or Tm6B ;
♂ 122 ;
C1-Gal4, UAS-mCD8-Cherry +
FRT82B, Gal80
FRT82B, Tsc1