fly-genetics-homework (2).doc

1Read the paper provided, and describe how the following experiments are accomplished:(1) To replace wildtype Wg with membrane-tethereda. Inject the posterior region of pre-cellular white1118embryos with a plasmid encoding Cas9 and another plasmid encoding a wingless specific targete gRNA, along with the pTVCherry wingless and a plasmid encoding FLP and I-Scel.b. The resulting adults were backcrossed to white1118 flies.c. Screening the subsequent F2 progeny for red eyes. The genotypes of these flies are as follows:The above three steps are as follows:d. Cre-mediated excision to produce wgKO：e. Confirm faithful expression of the reintegrate cDNA at the targeted locus：Insert a wild-type Wg cDNA into the attP site, along with mini-white as a genetic marker, and the resulting flies (wgKO; Wg) were indistinguishable from wild-type flies, confirming faithful expression of the reintegrated cDNAf. The reintegration of the NRTWg cDNA in wgKO to produce wgKO；NRT-Wg(2) To generate wg-/- clones in wildtype background or wg-/- clones in tethered wg backgrounda. To generate wg-/- clones in wildtype backgroundFRT40 wgCX4FRT40 wgCX4FRT40 ubiquitin-GFPFRT40 ubiquitin-GFPFRT40 ubiquitin-GFPFlipaseFRT40 wgCX4FRT40 ubiquitin-GFPFRT40 wgCX4FRT40 ubiquitin-GFPMitosisFRT40 wgCX4FRT40 wgCX4FRT40 ubiquitin-GFPFRT40 ubiquitin-GFPb. To generate wg-/- clones in tethered wg background FRT40 wgCX4FRT40 wgCX4FRT40 ubiquitin-GFP NRT-WgFRT40 ubiquitin-GFP NRT-WgFlipaseFRT40 wgCX4FRT40 ubiquitin-GFP NRT-WgFRT40 wgCX4FRT40 ubiquitin-GFP NRT-WgmitosisFRT40 wgCX4FRT40 wgCX4FRT40 ubiquitin-GFP NRT-WgFRT40 ubiquitin-GFP NRT-Wgc. To generate flies with wild-type wg in the body and tethered wg in the wings, or flies with tethered wg in the body and wildtype wg in the wings(a). To generate wgKO; WTBody; NRTWgDiscswgFRT Wg FRT NRTWgvestigial-gal4, UAS-Flpa.wgKO; WTBody; NRTWgDiscs (b). To generate wgKO; NRTWgBody;WTDiscswgFRT NRTWg FRT Wg vestigial-gal4, UAS-FlpwgKO; NRTWgBody;WTDiscs P esg-Gal4 / Cyo; Gal80ts UAS-IR / TM6 2. Design a simple and efficient genetic screen for tumor suppressors in the Drosophila digestive tract. 18Wild type F1 : esg-Gal4 / UAS-IR ; Gal80tsShift to 29 for 7 d 解剖F1 肠子，经固定、脱水、染色、制片，最后在荧光显微镜下观察肠道细胞的组成情况，如果有tumour产生，则说明相应的基因是tumour suppressor.Please send your homework in WORD or PDF format to and the email subject should be written as “Genetics Homework”. If you handwrite your answers or use schematic drawing in your answers and dig