The T315I mutation was introduced by site-directed mutagenesis.
Subsequently, a R419L mutation was introduced into purA to reduce the flux from IMP to AMP.
TA4135, into which this mutation was introduced, showed a same tendency of tolerance to other alcohols (ethanol and 1-butanol).
T-type currents recorded from tsA-201 cells expressing the rCav3.2 (+25) channel isoform were significantly increased when the GAERS mutation was introduced (Fig. 6D and E).
When the mutation was introduced into the encapsulated D39 strain (serotype 2), it led to attenuated virulence in mouse models either by intranasal colonization or by intraperitoneal infection.
When a mutation was introduced into the putative catalytic site for the xylanase domain (E280S), the ferulic acid esterase activity increased threefold, which suggests that the two catalytic domains for Xyn10D-Fae1A are functionally coupled.
In brief, the S89G mutation was introduced in exon 6 using an overlap extension-PCR method.
This mutation was introduced by overlapping two fragments amplified by primer sets mrarg5F and mrarg5iR and mrarg5if and mrarg5R.
The mutation was introduced into pET22b/ArgRS using the site-directed mutagenesis and purified these proteins with the same method as described previously.
Nucleotide sequencing of all PCR fragments cloned was carried out at BMR Genomics (Padova, Italy) to check whether any mutation was introduced.
Whole-genome resequencing revealed that no mutation was introduced into the plasmid, which was consistent with the unchanged MDH activity (Fig. 1b).
