(F ) A Hh reporter assay was used to measure signaling in Smo−/− cells transiently transfected with constructs encoding various mSmo point mutants and then treated (48 hr) with Shh, SAG (100 nM) or 20(S )-OHC (10 μM).
As we had already established that the FKH domain was dispensable for binding to c-Rel, we based the various deletion and point mutants on Foxp3ΔFKH (Fig. 2).
By using the Multi Site-Directed Mutasenesis Kit (Medical Biology Laboratemplate a template with wild-type hTFIIEα cDNA plasmid or a hTFIIH p62 cDNA mutant in which a NdeI site was disrupted by changing the third nucleotide of the 45th histidine codon of wild-type hTFIIH p62 cDNA, T to C, various oligonucleotide-mediated point mutants were created (Kunkel et al, 1987).
In order to define the molecular determinants of NR2B that govern con-G selectivity, we evaluated the ability of con-G to inhibit NMDAR ion channels expressed in human embryonic kidney (HEK 293 cells transfected with NR1, in combination with various NR2A/2B chimeras and point mutants, by electrophysiology using cells voltage-clamped in the whole-cell configuration.
Various studies using alanine point mutants and synthetic peptide scaffolds have collectively reported the importance of amino acid residues Q170, K171, E172, G173, W190, T195, and L196 of CCR5 EC2 in 2D7 recognition.
Different point mutants analyzed displayed various reduced levels of dNADH-K3Fe CN 6 dNADH-K3Fe CN 6ity.
Surprisingly, whole-genome sequencing analysis of 3 POA-resistant mutants revealed they had various point mutations (M117I, C deletion at nucleotide position 393, V138A mutations) in the panD gene (Table 1).
SEC and GTP binding by BBS1 and ARL6 point mutants.
Null mutants of each of the genes are viable, and have phenotypes similar to those of the point mutants.
Our results with the uvrB5 recF322::Tn3 DrecBC were similar to those we found using point mutants.
