Genetic analysis of dig-1
| Genotype . | n . | PVQ axon flip-over (% animals) . | Misplaced PVQ soma (% animals) . | n . | Misplaced amphid soma (% animals) . |
|---|---|---|---|---|---|
| Wild type | 399 | 8.3 | 0 | 150 | 0 |
| Complementation test with dig-1 | |||||
| dig-1(ky188) / dig-1(n1321) | 69 | n.d. | 46 | 44 | 64 |
| + / dig-1(ky188) | 23 | n.d. | 4.3 | 21 | 0 |
| + / dig-1(n1321) | − | n.a. | n.a. | 51 | 2 |
| dig-1(ky188) | 209 | 30.6 | 35.7 | 116 | 48 |
| dig-1(n1321) | 308 | 13 | 35.7 | 138 | 91 |
| Analysis over deficiency* | |||||
| dig-1(ky188) / nDf16 | 72 | 31 | 41 | 74 | 67 |
| dig-1(nu345) / nDf16 | 70 | 42 | 45.7 | 85 | 72.9 |
| + / nDf16 | 39 | 7.7 | 0 | 44 | 2.3 |
| dig-1(ky188) | 209 | 30.6 | 35.7 | 116 | 48 |
| dig-1(nu345) | 299 | 12 | 39.8 | 125 | 72 |
| Cosmid rescue | |||||
| dig-1(ky188); otEx2651 [K07E12,pRF4]† | 226 | 5.8 | 31.9 | − | n.a. |
| dig-1(ky188); otEx2110 [K07E12, R05H11,pRF4]‡ | 51 | 7.8 | 19.6 | − | n.a. |
| dig-1(ky188); rhEx40 [K07E12, R05H11, pRF4] | 183 | 8.7 | 6 | 170 | 17.6 |
| dig-1(nu336); rhEx40 [K07E12, R05H11, pRF4] | 206 | 2 | 8.7 | 127 | 16.5 |
| dig-1(n1480); rhEx40 [K07E12, R05H11, pRF4] | 332 | 7.8 | 5.1 | 68 | 13.2 |
| dig-1(ky188) | 209 | 30.6 | 35.7 | 116 | 48 |
| dig-1(nu336) | 212 | 11.3 | 44.4 | 74 | 66 |
| dig-1(n1480) | 174 | 6.3 | 41.4 | 101 | 52 |
| RNAi analysis | |||||
| rrf-3; RNAi (L4440 control)§ | 115 | 7 | 0 | 135 | 2.2 |
| rrf-3; RNAi(dig-1)§ | 155 | 31.5 | 17.3 | 205 | 76.6 |
| ky188; RNAi (dig-1) | 121 | 28.1 | 40.1 | 144 | 56 |
| nu345; RNAi (dig-1) | 163 | 35 | 49 | 153 | 69.2 |
| Genotype . | n . | PVQ axon flip-over (% animals) . | Misplaced PVQ soma (% animals) . | n . | Misplaced amphid soma (% animals) . |
|---|---|---|---|---|---|
| Wild type | 399 | 8.3 | 0 | 150 | 0 |
| Complementation test with dig-1 | |||||
| dig-1(ky188) / dig-1(n1321) | 69 | n.d. | 46 | 44 | 64 |
| + / dig-1(ky188) | 23 | n.d. | 4.3 | 21 | 0 |
| + / dig-1(n1321) | − | n.a. | n.a. | 51 | 2 |
| dig-1(ky188) | 209 | 30.6 | 35.7 | 116 | 48 |
| dig-1(n1321) | 308 | 13 | 35.7 | 138 | 91 |
| Analysis over deficiency* | |||||
| dig-1(ky188) / nDf16 | 72 | 31 | 41 | 74 | 67 |
| dig-1(nu345) / nDf16 | 70 | 42 | 45.7 | 85 | 72.9 |
| + / nDf16 | 39 | 7.7 | 0 | 44 | 2.3 |
| dig-1(ky188) | 209 | 30.6 | 35.7 | 116 | 48 |
| dig-1(nu345) | 299 | 12 | 39.8 | 125 | 72 |
| Cosmid rescue | |||||
| dig-1(ky188); otEx2651 [K07E12,pRF4]† | 226 | 5.8 | 31.9 | − | n.a. |
| dig-1(ky188); otEx2110 [K07E12, R05H11,pRF4]‡ | 51 | 7.8 | 19.6 | − | n.a. |
| dig-1(ky188); rhEx40 [K07E12, R05H11, pRF4] | 183 | 8.7 | 6 | 170 | 17.6 |
| dig-1(nu336); rhEx40 [K07E12, R05H11, pRF4] | 206 | 2 | 8.7 | 127 | 16.5 |
| dig-1(n1480); rhEx40 [K07E12, R05H11, pRF4] | 332 | 7.8 | 5.1 | 68 | 13.2 |
| dig-1(ky188) | 209 | 30.6 | 35.7 | 116 | 48 |
| dig-1(nu336) | 212 | 11.3 | 44.4 | 74 | 66 |
| dig-1(n1480) | 174 | 6.3 | 41.4 | 101 | 52 |
| RNAi analysis | |||||
| rrf-3; RNAi (L4440 control)§ | 115 | 7 | 0 | 135 | 2.2 |
| rrf-3; RNAi(dig-1)§ | 155 | 31.5 | 17.3 | 205 | 76.6 |
| ky188; RNAi (dig-1) | 121 | 28.1 | 40.1 | 144 | 56 |
| nu345; RNAi (dig-1) | 163 | 35 | 49 | 153 | 69.2 |
All strains [except + / dig-1(n1321)] contain the oyls 14(sra-6::gfp) transgene in the background to visualize PVQ anatomy. Amphid neuron cell body placement was assayed by Dil staining.
CX2914 nDf16/dpy-17 unc-32 hermaphrodites produce progeny of the expected phenotypes in skewed proportions, revealing haplo-insufficiency of some gene(s) deleted by nDf16 (41% dead embryos, 24% wild-type and 35% Dpy Unc found in the brood of seven nDf16/dpy-17 unc-32 animals). ky188; oyls14 homozygous males were mated into nDf16/dpy-17 unc-32 hermaphrodites. Most F1 cross progeny were ky188/dpy-17 unc-32, with few ky188/nDf16 produced (due to haplo-insufficiency). Singled ky188/nDf16 F1 cross progeny (no Dpy Unc worms in the F2 brood), were examined as 3-4 day old adults. The entire F2 brood was inspected for the presence of abnormal larvae and the proportion of dead embryos was counted. To increase the number of observations, numerous F2 worms were singled from ky188/nDf16 F1 mothers, and those yielding dead embryos in the F3 generation were examined at the neuroanatomical level. The analysis was performed in the same way for nu345;oyls14, and the oyls14 control.
Data for one rescuing line is shown. 2/3 lines rescued ky188defects. Cosmid K07E12 was injected at ~10 ng/μL along with pRF4 (150 ng/μL) as injection marker.
Data for one rescuing line is shown. 4/5 lines rescued ky188defects. Cosmids were injected at ~10 ng/μL along with pRF4 (150 ng/μL)as injection marker.
RNAi was performed in a genetically sensitized, rrf-3 mutant background (Simmer et al.,2002).