TABLE 1.
Molecular analyses of D1EP473 w− excision lines
| Linea | D1 proximal + Pry4 PCR | D1 distal + Plac1 PCR | D1 proximal + D1 distal PCRb | Southernc |
|---|---|---|---|---|
| D1EP473 | 685 bp | 503 bp | 794 bp | ∼9.7 kb/∼1.7 kb |
| 1A | − | − | + | ∼1.3 kb/∼1.7 kb |
| 1B, 1C, 1D, 1F | − | − | + | ∼1.7 kb |
| 1E | − | − | +/∼0.85 kb | ∼1.7 kb doublet |
| 2A | + | − | + | ∼3.2 kb/∼1.7 kb |
| 2B, 2C | − | − | +/∼0.85 kb | ∼1.7 kb doublet |
| 3A | − | − | +/∼0.85 kb | ∼1.7 kb doublet |
| 4A | − | − | + | ∼1.7 kb |
| 70-1, 70-2, 70-3 | − | − | + | ∼1.7 kb |
| 70-5 | + | + | + | ∼9.7 kb/∼1.7 kb |
| 70-6 | − | − | +/∼0.85 kb | ∼1.7 kb doublet |
| 70-7 | − | − | + | ∼1.1 kb/∼1.7 kb |
| 70-9 | − | − | + | ∼1.7 kb |
The PCR primer sets are as illustrated in Figure 1. The presence of a PCR product of size expected for the original D1EP473/TM3, Sb strain (first row) is denoted by a +, the absence of a product by a −, and a product of other size by the estimated size.
Lines that may not be independent isolates are grouped together in a row. Analyses were performed on balanced lines, due to extraneous lethal mutations.
The PCR products derived from one or both homologs. When the intact P{EP} element is present, size limitations preclude amplification of a product from that homolog. However, the wild-type D1 locus of the balancer chromosome yielded a 794-bp product.
The D1 proximal + D1 distal PCR product was used as a probe of BamHI-digested genomic DNA, as illustrated for representative lines in Figure 4. The band(s) derived from one or both homologs. The intensity of the 1.7-kb band for lines 1A, 2A, 4A, 70-5, and 70-7 appeared approximately half that of the 1.7-kb band for all other w− lines.