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Wild-type and mutant sequences were compared to identify putative resistance mechanisms.
The mutant sequences were used in the mutant 3′-UTR constructs.
F3 and F5 mutant sequences were used for Flpe-mediated cassette exchange, and m2 and lox2272 mutant sequences were used for Cre-mediated cassette exchange due to their high incompatibility with wild-type sequences.
Sequencing of cDNA from cells infected with this last mutant demonstrated that the parental mutant sequence was retained and that genotypic revertants to the wild-type as well as new mutant sequences were generated.
All mutant sequences were verified by DNA sequencing.
However, even under selective pressure from monoclonal antibodies, very few escape mutant sequences were observed for M2e [30] suggesting that this region is biologically constrained.
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The mutant sequences are numbered from 1 to 12. HAuCl4, sodium tris-citrate, NaOH and HCl were purchased from Xilong Chemical (Guangdong, China).
Wild type and mutant sequences are shown at the bottom.
The corrected wild-type and mutant sequences are shown in Figure 7.
Primer sets used to produce MLK3 mutant sequences are described in Supplementary Material, Table S2.
The wild-type and mutant sequences are separated by the YIP vector sequences including a selectable marker [ 11].
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