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Future research should concentrate on specific indica markers targeting the identified insertion in the Pi33 zone.
The TnPZ locations in these 35 strains were identified by PCR with primers specific for already identified insertion sites, and by direct chromosomal sequencing from element ends when needed.
Using cross-species comparisons and the identified insertion positions, a presence absence matrix was created for approximately 796,000 insertions.
No genes were surrounded on both sides by IS but, since no recent HGT was identified, insertion sequences might have diverged over time sufficiently to impair their identification.
Taking the above observations together we identified insertion biases, gene expression and protein structural constraints as likely limiting factors for the success of protein trap screens.
Though precise parallel insertions cannot be completely ruled out, mapping accuracy is strong evidence that identified insertion positions within a narrow window are likely shared, orthologous insertions acquired from the common ancestor.
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We also identified insertions in hemY (SAOUHSC_01460), a gene involved in the production of protoheme [ 84].
Remarkably, we also identified insertions and deletions in four different amino acid repeats present in this gene (Table 5).
We used the presence or absence of all identified insertions to calculate frequency distributions for all three species.
Using TAIL-PCR we identified insertions in parasites of parent population 2 and 5 and subpopulations of P1, P2 and P5.
We examined the nucleotide sequence corresponding to the exons of the parent gene and identified insertions, deletions, and base pair substitutions.
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