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Ribosome-mediated polymerization of backbone-extended monomers into polypeptides is challenging due to their poor compatibility with the translation apparatus, which evolved to use α- L -amino acids. Moreover, mechanisms to acylate (or charge) these monomers to transfer RNAs (tRNAs) to make aminoacyl-tRNA substrates is a bottleneck. Here, we rationally design non-canonical amino acid analogs with extended carbon chains (γ-, δ-, ε-, and ζ-) or cyclic structures (cyclobutane, cyclopentane, and cyclohexane) to improve tRNA charging. We then demonstrate site-specific incorporation of these non-canonical, backbone-extended monomers at the N- and C- terminus of peptides using wild-type and engineered ribosomes. This work expands the scope of ribosome-mediated polymerization, setting the stage for new medicines and materials.
Ribosome-mediated polymerization of backbone-extended monomers into polypeptides is challenging due to their poor compatibility with the translation apparatus, which evolved to use α- L -amino acids. Moreover, mechanisms to acylate (or charge) these monomers to transfer RNAs (tRNAs) to make aminoacyl-tRNA substrates is a bottleneck. Here, we rationally design non-canonical amino acid analogs with extended carbon chains (γ-, δ-, ε-, and ζ-) or cyclic structures (cyclobutane, cyclopentane, and cyclohexane) to improve tRNA charging. We then demonstrate site-specific incorporation of these non-canonical, backbone-extended monomers at the N- and C- terminus of peptides using wild-type and engineered ribosomes. This work expands the scope of ribosome-mediated polymerization, setting the stage for new medicines and materials.
Fatigue crack initiation and propagation behaviour of CFRP have been of great importance because such composites are often used in engineering components that are subjected to continuous cyclic loading. The objective of this thesis work was to investigate the damage characteristics of the fatigue properties of CFRP composites by the modification of the polymer matrix with nanoclay addition. Carbon fibre reinforced epoxy was produced via vacuum assisted resin infusion moulding method (VARIM) with nanoclay concentrations of 0wt%, 1wt%, 3wt% and 5wt%. Tension-tension fatigue tests were conducted at loading levels of 90%, 75% and 60%. The frequency that was used was 3Hz with R value of 0.1. The results showed that at nanoclay percentages of 0wt%, 1wt% and 3wt% there was a consistent trend, where the number of cycles increased in fatigue loading percentages of 90%, 75% and 60%. At 5wt% nanoclay percentage the number of fatigue cycles dropped significantly at the 90% fatigue loading. The brittle nature of the 5wt% laminate became dominate and the sample fractured early at low fatigue cycle numbers. At the 75% fatigue loading, the number of cycles increased and at 60% fatigue loading the 5wt% nanoclay sample exceeded the number of cycles of all the nanoclay percentages by 194%. This was due to the intercalated arrangement of the nanoclays favouring the slow rate of surface temperature increase, during fatigue testing, at low fatigue cycle loading. The Crack Density analysis was performed and showed that at the same time in the fatigue cycle life, the 1wt% had 55 cracks, 3wt% had 52 cracks and the 5wt% had 50 cracks, for the 60% fatigue loading. This proved that it took longer for the cracks to initiate and propagate through the sample as the nanoclay percentage increased. Impact and hardness testing showed that the 5wt% exhibited brittle behaviour, which contributed to the results above. Scanning electron microscopy examination highlighted that the agglomeration of nanoclays delayed the crack initiation and propagation through the specimen and that the extent of fatigue damage decreased as the nanoclay percentage increased. A fatigue failure matrix was developed and showed that delamination, fibre breakage and matrix failure were the predominate causes for the fatigue failure.
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