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1) Product Images from "CynD, the Cyanide Dihydratase from Bacillus pumilus: Gene Cloning and Structural Studies"
Article Title: CynD, the Cyanide Dihydratase from Bacillus pumilus: Gene Cloning and Structural Studies
Journal: Applied and Environmental Microbiology
Figure Legend Snippet: Homologous recombination in B . pumilus C1 using p3022. The hybrid plasmid p3022 was made by cloning p3019 into the Xba I and Pst I sites of pE194. This plasmid was designed to be used as a temperature-sensitive shuttle plasmid for Bacillus and E . coli . The bla gene encodes ampicillin resistance for selection in E . coli , and ermC encodes erythromycin resistance for selection in Bacillus . The cynD 539 encodes the partial 539-bp PCR product from the putative cyanide dihydratase of B . pumilus C1. B . pumilus C1 was transformed with p3022 to make strain MB3025. This diagram shows p3022 integration into the cyanide dihydratase gene of B . pumilus C1 by homologous recombination. (A) Recognition of homologous regions of p3022 with the gene encoding cyanide dihydratase. (B) The integrated plasmid p3022 flanked by the gene encoding cyanide dihydratase. The gene encoding replication initiation protein for plasmid replication in Bacillus species, repF , is also shown. The origin of plus-strand synthesis, ori + , is not shown but is located within the repF gene.
Techniques Used: Homologous Recombination, Plasmid Preparation, Clone Assay, Selection, Polymerase Chain Reaction, Transformation Assay
Figure Legend Snippet: Three-dimensional reconstruction of negatively stained micrographs of the cyanide dihydratase from B . pumilus at a resolution of 3.2 nm. Shown are the views down the twofold axis (A and C) and a view perpendicular to the twofold axis (B). The structure is a spiral of two turns from end to end, having 18 subunits.
Techniques Used: Staining
Figure Legend Snippet: Negative-stain electron micrographs of the isolated cyanide dihydratase from B . pumilus at pH 8.0 (A) and 5.4 (B) illustrating the transition from discrete particles to extended rods. (C) Platinum-carbon shadowing of the cyanide dihydratase rods. Regular striations along the length of the rod can be seen in rods lying parallel to the direction of shadowing. The angle of the striations implies that the helical arrangement of subunits is left handed. The location of five of the striations is emphasized by white lines.
Techniques Used: Staining, Isolation