First, TAGIT includes the lacZ gene to allow the rapid identification of in-frame insertions and significantly reduce the number of insertions screened.
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.
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).
Since NHEJ is a fairly common phenomenon in Kluvyveromyces marxianus, random gene insertion and screening for activity might be a good strategy for host engineering [ 23].
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).
Mutants identified to be hypercytotoxic in vitro were more likely to be avirulent (12 of 28 strains, 43%) as compared to transposon insertion mutations screened at random for virulence in which 4 6% of insertion mutations were avirulent [21].
PROGINS insertion was screened by electrophoresis.
Insertion was screened by PCR and fluorescence visual inspection.
All P-element insertion lines screened were GS (two UAS sequences in opposite directions [60]) or NP (Gal4 enhancer trap; [61]).
