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To make LMB-2 less immunogenic, we introduced a large deletion in domain II and six point mutations in domain III that were previously shown to reduce T cell activation in other RITs.
Loss of proteolytic function was limited to mutations in Domain 1 in the N-terminal third of the EspP passenger.
It is also striking that genetic screens have never identified fidelity mutations in domain 3, which is also proposed to interact with aa-tRNA.
Previous work has demonstrated the importance of domains 1 and 3a of Munc18-1 in closed-conformation syntaxin binding [9] and specific point mutations in domain 1 can inhibit syntaxin binding [11], [29], [30].
However, neither of the two mutations in domain II of Cry8Ka5 is located in these loops.
None of the strains had any mutations in domain II of 23S rRNA.
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Novel chaperone molecules should not only be advantageous in terms of improved enhancing profile, but may also increase the rate of responsive mutations by assisting the folding of enzymes with mutations in domains other than the catalytic site.
Furthermore, a cDNA variant encoding a point mutation in domain 11 (Ile→Thr) was generated (M6P/IGF2R DoM6P/IGF2R
The snpA6 suppressor was shown to involve an E117K mutation in domain 1 eRF1 of a UGA stop codon in ngA1.
76 isolated M. pneumoniae strains and the standard strain FH all showed no gene mutation in domain II of 23S rRNA.
It's worthy to note that the most common mutation in domain V of the 23S rRNA gene, G2576T, was also not detected in our 5 LRSC isolates, whereas, G2603T was detected in all these 5 isolates.
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