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In a large retroviral insertion screen in mice [ 11- 14], we have identified Gpr110 as an oncogene.
A transposon insertion screen to identify mutants defective in SOS induction following nalidixic treatment returned 18 mutants, all of which were recB or recC mutants (Newmark et al. 2005).
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P element insertion screens allow easier identification of mutated genes, but P element mutations are usually not nulls, and on average have weaker embryonic phenotypes than EMS mutations in the same genes.
First, TAGIT includes the lacZ gene to allow the rapid identification of in-frame insertions and significantly reduce the number of insertions screened.
Existing methods are time consuming and expensive since they involve post-insertion screening steps.
Mos-mediated single-copy transgenes were created as previously described, utilizing the ttTi5605 Mos site on chromosome II and the direct-insertion screening method (Frokjaer-Jensen et al., 2008).
However, in a large insertion mutagenesis screen, a zebrafish mutant harboring an insertion in the first intron of cofilin 1 (cfl1) (previously known as cofilin 2 like) was identified and showed no obvious phenotype at one day after fertilization [30].
In a retroviral insertion mutagenesis screen in the mouse, we identified GPR110 as an oncogene.
We began with a transposon insertion mutagenesis screen (Burns et al. 1994; Kumar and Snyder 2002) in which we tested 9000 independent transformants for improved or impaired growth compared with the spt10 Δ parent (Materials and Methods).
PROGINS insertion was screened by electrophoresis.
Insertion was screened by PCR and fluorescence visual inspection.
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