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in Wiley InterScience (www.interscience.wiley.com).We developed a method for coupling protein to its coding DNA on magnetic microbeads using emulsion PCR and cell-free protein synthesis in emulsion. A PCR mixture containing streptavidin-coated microbeads was compartmentalized by water-in-oil (w/o) emulsion with estimated 0.5 template molecules per droplet. The template molecules were amplified and immobilized on beads via bead-linked reverse primers and biotinylated forward primers. After amplification, the templates were sequentially labeled with streptavidin and biotinylated anti-glutathione S-transferase (GST) antibody. The pool of beads was then subjected to cellfree protein synthesis compartmentalized in another w/o emulsion, in which templates were coupled to their coding proteins. We mixed two types of DNA templates of Histidine6 tag (His6)-fused and FLAG tag-fused GST in a ratio of 1:1,000 (His6: FLAG) for use as a model DNA library. After incubation with fluorescein isothiocyanate (FITC)-labeled antiHis6 (C-term) antibody, the beads with the His6 gene were enriched 917-fold in a singleround screening by using flow cytometry. A library with a theoretical diversity of 10 6 was constructed by randomizing the middle four residues of the His6 tag. After a two-round screening, the randomized sequences were substantially converged to peptide-encoding sequences recognized by the anti-His6 antibody.

SummaryThe inhibitory effect of dietary taurine on cholesterol gallstone formation was studied using male mice of Jcl: ICR strain. Mice were provided three kinds of semi-purified diet; a cholesterol-free diet (standard), a lithogenic diet containing 0.5% cholesterol and 0.25% sodium cholate (C-CA), and a lithogenic diet supplemented with 5% taurine. The changes of total cholesterol in serum, cholesterol mass in the liver and the gall bladder, and gallstone incidence were studied as a function of time. Gallstone formation was observed only in the mice fed on the C-CA diet for more than 3 weeks. The changes of serum cholesterol concentration were not consistent with gallstone formation. The choles terol mass of the liver in taurine-supplemented mice decreased after the 3rd week. Cholesterol content of the gall bladder increased with choles terol gallstone formation. Therefore, the inhibitory effect of dietary taurine on cholesterol gallstone formation may be related to the decrease in cholesterol content of the liver.

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