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In the dark, one big group, which showed no reaction at all using prolonged times, is composed of L112C-A132C/Basal mutant except for E113C/Basal mutant.
As shown in Table 1, most of the purified-mutants/Basal mutant except for P107C/, N111C/, G114C/, A124C/, L125C/, W126C/, L128C/, and E134C/Basal mutant formed the typical rhodopsin-like chromophore with A280/A500 ratios between 1.7 and 2.5, with an absorption λmax in the visible range varying from 480 nm to 496 nm.
To further examine whether one of more of the 18 remaining genomic changes observed in the HI mutant (except for the hit locus) are linked to the HI phenotype, we randomly selected one of the 6 isolates in which no hit mutation was found.
Typically, 6 L were expressed for each mutant, except for G117E (12 L).
It has been assumed that pyrimethamine resistance conferred by multiple mutations arose through stepwise selection of the S108N single mutant (except for the A16V/S108T allele).
RT-PCR did not detect any transcripts of MpSIG1 in the mutant except for a 1,493-bp 1,493-bproduct that was not present in the WT productfig. 1 D).
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For mutants L112C-A132C/Basal mutant, excepting E113C/Basal mutant, the lack of reactivity in the dark and the accessibility of cysteine after photoactivation indicate the increases of water/4-PDS accessibility for certain cys-mutants.
Likewise, a hypothetical gene (551.m00236, TGME49_108060) is highly expressed in all mutant lines except for mutants B7 and P11 (blue bars); an example of a candidate gene for future study.
All the mutants except for D150N completely lost their methylation activities.
Interestingly, using COS-1 cells as a eukaryotic expression system, all mutants except for those targeting p.L34 were found to be synthesized and secreted to the cell supernatant, from which they could be purified for further experiments [31],[32].
Soluble sugar levels in leaf sheaths were similar between lse mutants and WT, or were lower in the mutants, except for the fourth leaf sheath of lse2, in which sucrose levels were significantly higher than those of WT in both morning and evening.
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