For each session, we used traffic density, the most spatially dispersed indicator, to draw a stratified-random sample (without replacement) of six sites (e.g., randomly allocate 3 'high' and 'low' traffic density sites per session).
The plasmid pSKDuet16 contains an additional two mutations of HM to LG at the front of GB1 due to the replacement of NdeI site by AvrII site, compared with the plasmids derived from pJJDuet30 [8].
The mutations designed included replacement of cleavage sites with LVPRGS, a well-known thrombin substrate sequence, and insertion of this sequence near cleavage sites with appropriate mutations of the cleavage site.
The DNA base excision repair (BER) pathway is involved in maintaining DNA integrity through the removal and replacement of abasic sites (Evans et al, 2000).
The alkylation resulting from mutational replacement of active site residues or substitution of structurally diverse substrates was investigated in other sesquiterpene synthases possessing varying levels of product complexity.
Transformation of the CORE6 cassette into yeast and homologous recombination at the target site results in the replacement of the target site by the CORE6 cassette.
This particular phenomenon was induced by replacement of the Zn sites, instead of the O ones, with Sb atoms (Sb_Zn) to form a complex with two V_Zn, which is the Sb_Zn-2V_Zn complex.
To construct expression plasmids of cat1+ and its point mutants, cat1+ ORF amplified by PCR using a genomic library (NBRP) as a template was cloned into pREP273-EGFP, which was generated by the replacement of multi-cloning sites together with the nmt41 promoter (Basi et al., 1993) and 3HA fragment from pSLF273 (NBRP) and the insertion of EGFP ORF in NotI and BglII sites.
Contrary to this prediction we find that the rate of substitution at replacement sites remains greater (median dN = 0.01) than that observed across the UCP2 (median dN = 0.003, p-value < 0.001) and UCP3 (median dN = 0.004, p-value = 0.0365) phylogenies.
It remains to be seen, therefore, whether the reported reactivation by zinc is due to replacement of an active site metal, binding of zinc to an alternative site such as the polyhistidine expression tag, or an experimental artifact.
