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Analysis of novel alleles of brother of tout‐velu , the drosophila ortholog of human EXTL3 using a newly developed FRT42D ovo D chromosome
Author(s) -
Lujan Ernesto,
Bornemann Douglas J.,
Rottig Carmen,
Bayless Brian A.,
Stocker Hugo,
Hafen Ernst,
Arora Kavita,
Warrior Rahul
Publication year - 2016
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.22981
Subject(s) - biology , genetics , germline , in ovo , mutant , somatic cell , mitotic crossover , chromosome , mutation , germline mutation , allele , phenotype , gene , microbiology and biotechnology , embryo
The FLP/FRT system permits rapid phenotypic screening of homozygous lethal mutations in the context of a viable mosaic fly. Combining this system with ovo D 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 ( neo R ) transgenes are in common use. Parallel ovo D lines were developed using w + mW.hs FRT insertions on the X and chromosomes 2R and 3L, as well as ry + , neo R FRT insertions on 2L and 3R. Consequently, mutations isolated on the X, 2R and 3L chromosomes in a ry + , neo R 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 ovo D line for the ry + , neo R 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.