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The knockout obtained was non-polar.
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Since Southern blot analysis revealed that the knockout lines obtained had multiple insertions of the construct (data not shown), one knockout line (Ppα-dox-2) with null Ppα-DOX activity was selected.
Genes coding for DUF-HIC proteins were over-represented in the lethal or medium-dependent knockouts obtained since five of the six genes involved belong to this class (p = 0.009; Fisher's exact test).
BY4742 (MATα his3 Δ1 leu2 Δ0 lys2 Δ0 ura3 Δ0) was used for transceptor work as wild-type and gap1 Δ and pho84 Δ mutants made in BY4742 were from the yeast knockout collection obtained from Open Biosystems, with either gene deleted by KanMX.
Figure 4 illustrates a dendrogram of the production targets, in which the similarity between two targets has been calculated by the membership-based Jaccard similarity coefficient (Levanodowsky and Winter, 1971) of the reaction knockouts sets obtained by FastPros-based OptKnock screening.
All knockout strains obtained in the present study were confirmed by PCR experiments.
We obtained the knockout fitness data from Segre et al. (2005) and inspected the epistasis status of the three WGD pair groups (CF, NF, and SF).
The obtained knockout mutant was designated as T. aotearoense LA1002.
Importantly, biallelic gene knockout is obtained at high frequency by only one round of targeting using a single marker.
We first tested murine embryonic fibroblasts (MEFs) from src-/-yes-/-fyn-/ triple knockout mice (obtained from ATCC).
During the process of making deletion mutants in hrdC in the other two strain backgrounds, we also noticed a low homologous recombination frequency to obtain the knockout strains.
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