Analysis of novel alleles of brother of tout-velu, the drosophila ortholog of human EXTL3 using a newly developed FRT42D ovoD chromosome

• Published in: Genesis (New York, N.Y. : 2000) N.Y. : 2000), 2016-11, Vol.54 (11), p.573-581
• Main Authors: Lujan, Ernesto, Bornemann, Douglas J., Rottig, Carmen, Bayless, Brian A., Stocker, Hugo, Hafen, Ernst, Arora, Kavita, Warrior, Rahul
• Format: Article
• Language: English
• Subjects: botv; brother of tout-velu; Chromosomes; Drosophila FRT42D ovoD; EXTL3; germline clonal analysis; heparan sulfate proteoglycan; HSPG; Mutation; N-acetylglucosamine transferase-I/II
• ISSN: 1526-954X; 1526-968X
• DOI: 10.1002/dvg.22981

The FLP/FRT system permits rapid phenotypic screening of homozygous lethal mutations in the context of a viable mosaic fly. Combining this system with ovoD dominant female‐sterile transgenes enables efficient production of embryos derived from mutant germline clones lacking maternal contribution from a gene of interest. Two distinct sets of FRT chromosomes, carrying either the mini‐white (w + mW.hs), or rosy (ry+) and neomycin (neoR) transgenes are in common use. Parallel ovoD lines were developed using w + mW.hs FRT insertions on the X and chromosomes 2R and 3L, as well as ry+, neoR FRT insertions on 2L and 3R. Consequently, mutations isolated on the X, 2R and 3L chromosomes in a ry+, neoR FRT background, are not amenable to germline clonal analysis without labor‐intensive recombination onto chromosome arms containing a w + mW.hs FRT. Here we report the creation of a new ovoD line for the ry+, neoR FRT insertion at position FRT42D on chromosome 2R, through induced recombination in males. To establish the developmental relevance of this reagent we characterized the maternal‐effect phenotypes of novel brother of tout‐velu alleles generated on an FRT42D chromosome in a somatic mosaic screen. We find that an apparent null mutation that causes severe defects in somatic tissues has a much milder effect on embryonic patterning, emphasizing the necessity of analyzing mutant phenotypes at multiple developmental stages.