Variant d5f encodes a non-functional isoform of chTERT and i10ec, generating a variant targeted by the NMD pathway.
Where available, candidate DNA variant positions were annotated with RNA information by generating a pileup of the DNA variant position in the RNA BAM alignment file.
We envision that this could be achieved by either increasing MOB1 phosphorylation by MST1/2, known to increase LATS1/2-MOB1 interactions[105], or by generating a MOB1-independent active LATS variant.
Hence, we applied either a Chi-square test or Fisher's exact test to each identified variant by generating a two by two table, where the rows are labeled as the forward or the reverse strand, and the columns are labeled as the target allele and other alleles, and each entry of the cell registers the corresponding count.
We began by generating a pool of protein 18-19 variants using error-prone PCR to produce random point mutations that would lead to single-residue changes in the protein [14].
cDNA encoding wild-type human ABCB6 (NM_005689) [ 12] was mutated by PCR to generate a variant bearing a human influenza haemagglutinin (HA) or FLAG tag at its C-terminus.
They further exhibited the crucial role of the tyrosine and glutamine residues (loop 3) by generating mutated variants (Y replaced by A, S or F; Q was replaced by A or E), all of which were inactive or did not bind to the receptor (Koehbach et al., 2013).
The entrance to the active site was confirmed by generating variant G255F which exhibited only 7% of the wild-type's specific activity of product formation, suggesting inhibition of substrate entrance into the active site by the large aromatic residue.
In conclusion, this work allowed introducing random diversity in the gene coding for the cellulase from Streptomyces sp. G12 expressed in E. coli rCelStrep by epPCR, generating a library of 10,000 variants that were screened by an ad hoc strategy of automated HTS for variants with higher cellulolytic activity.
Diminished APOBEC activity and sublethal levels of hypermutation may favor virus evolution, by generating viable variants carrying immune escape or drug resistance mutations.
