36 research outputs found
Characterisation of a Desmosterol Reductase Involved in Phytosterol Dealkylation in the Silkworm, Bombyx mori
Most species of invertebrate animals cannot synthesise sterols de novo and many that feed on plants dealkylate phytosterols (mostly C29 and C28) yielding cholesterol (C27). The final step of this dealkylation pathway involves desmosterol reductase (DHCR24)-catalysed reduction of desmosterol to cholesterol. We now report the molecular characterisation in the silkworm, Bombyx mori, of such a desmosterol reductase involved in production of cholesterol from phytosterol, rather than in de novo synthesis of cholesterol. Phylogenomic analysis of putative desmosterol reductases revealed the occurrence of various clades that allowed for the identification of a strong reductase candidate gene in Bombyx mori (BGIBMGA 005735). Following PCR-based cloning of the cDNA (1.6 kb) and its heterologous expression in Saccharomyces cerevisae, the recombinant protein catalysed reduction of desmosterol to cholesterol in an NADH- and FAD- dependent reaction
Non-origami DNA for functional nanostructures: From structural control to advanced applications
Base-pair mapping by chemical force microscopy on nucleobase self-assembled monolayers
Self-assembled monolayers (SAMs) of double-chain disulfide derivatives of nucleobases (adenine and thymine) were formed on Au substrates in order to measure complementary hydrogen bonding by chemical force microscopy at the interfaces. Surface plasmon resonance measurements indicated that the formation of the nucleobase SAMs on Au surface was completed within 80 min. To measure adhesion force by atomic force microscopy (AFM), Au- coated AFM tips were modified with the nucleobase SAMs, too. SAM-modified An substrates micro-patterned on quartz substrates were prepared for adhesion force mapping. The adhesion force between the complementary nucleobases is larger than that of the non-complementary combination. Electrochemical detection using a redox-intercalator was demonstrated for hybridization of single-stranded polynucleic acid with the nucleobase SAMs modified on Au electrodes. (C) 2002 Elsevier Science B.V. All rights reserved