In theory, linear substrates may be as efficient substrates as their circular counterparts but the outcome of vector incorporation into the genomic FRT site may differ.
By trapping the first and last acyl-thioesterase intermediates in the catalytic cycle as DAP conjugates, we provide structural insight into how conformational changes in thioesterase domains of such nonribosomal peptide synthetases control the oligomerization and cyclization of linear substrates.
lin, linear substrate DNA; oc, open-circular products; scc, supercoiled closed-circular products; mult, dimeric and higher-order multimeric products.
Our attribution of this effect to ks-1→p (and not kcl or klig) is supported by previous observations that cleavage (kcl) and ligation (klig) are similar in linear and supercoiled substrates.
Non-viral vectors were immobilized to substrates in linear patterns using microfluidic techniques, and cells cultured on the surface had localized gene expression within the cell population.
The relative importance of soil layer, geographic region, total carbon, CAZy composition and community structure in linear models for each substrate was assessed using the R package relaimpo using the primary and secondary axes of NMDS as measures of community structure and CAZy composition.
Transposon excision and in vitro transposition occurred only when the transposase acted on its own IR sequence; we observed minimal cross-recognition of the transposon ends in cleavage assays using linear DNA substrates or plasmid substrates.
Finally, we investigated the effects of rate laws with non-linear substrate dependencies in the absence of cofactors.
This poses problems for the definition of reactant pairs based on compound similarity (see Section 2.2), particularly if a substrate is in linear form and the corresponding product in ring form.
A comparison to related enzymes suggests that this form of substrate conformational change likely represents a common feature of Schiff base formation in enzymes acting on linear ketone substrates.
