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As shown in Figure 4B and C, both constructs were modified by Ubc13 Mms2 Rad5.
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These constructs were modified by using QuickChange II Site-Directed Mutagenesis Kit (STRATAGENE, La Jolla, CA) to generate a point mutation at −358 from G to A. The primers used were 5'-CTTACTGTCTGCTCAGGAGCTTCCTCTTGGCCCCG, and 5'- CGGGGCCAAGAGGAAGCTCCTGAGCAGACAGTAAG.
However, the two constructs were modified with very different efficiencies.
As the pyramid identified needs, some constructs were modified to convey a positive meaning ('fatigue' was for instance modified to 'energy'energy
The donor constructs were modified to include two NotI sites so that a straightforward digestion step could be used to reduce plasmid-transposon junction background.
The GST-CNB-1 and TAX-6 bacterial expression constructs were modified from the GST-CNB-1 and GST-TAX-6 plasmids reported before (Bandyopadhyay et al., 2002).
N-terminally 6×-His tagged wild-type (WT) HγD-Crys expression constructs were modified via site-directed mutagenesis to introduce Y A, F A, and C S substitutions as described previously.
Specifically, the TyrTCR construct was modified to express murine TCR constant regions and human TCR variable regions [34].
The murine TyrTCR expression construct was modified for optimized surface expression and improved tetramer reactivity (R. Koya, unpublished observations).
The construct was modified to contain a FLAG tag (DYKDDDDK).
Additionally, the 2.7hPLPZ construct was modified to generate two plasmids, each having a unique BstBI site.
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