Download Applications of Chimeric Genes and Hybrid Proteins by Thorner J. (ed.), Emr S.D. (ed.), Abelson J.N. (ed.) PDF

By Thorner J. (ed.), Emr S.D. (ed.), Abelson J.N. (ed.)

The severely acclaimed laboratory average for greater than 40 years, tools in Enzymology is without doubt one of the so much hugely revered guides within the box of biochemistry. seeing that 1955, every one quantity has been eagerly awaited, usually consulted, and praised by means of researchers and reviewers alike. Now with greater than three hundred volumes (all of them nonetheless in print), the sequence comprises a lot fabric nonetheless correct today-truly a vital book for researchers in all fields of lifestyles sciences.

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Coli ALKALINEPHOSPHATASEGENE FUSIONS 35 expression of the downstream gene lacY. This permits an enrichment of interesting nonpolar, Lac-down mutations with specific effects on the fusion protein from the much larger class of nonsense mutations that simply abolish expression of both lacZ and lacY. 3° Conclusion Despite the usefulness of new reporter systems such as GFP, and the many epitope-tagging systems, the Lac system remains a powerful genetic tool both in studying gene regulation and in generating mutants in diverse complex biological systems.

Fluorescent Antibody Labeling for Flow Cytometry Strains and Reagents. , FACScan or FACScalibur); FACS analysis tubes: 12 x 75-mm round-bottom tubes. Phosphate-buffered saline (PBS); antibody (Ab)-staining buffer [5% bovine serum albumin (BSA) in PBS]; midlog cultures of bacteria; Ab 20 R. H. Valdivia and S. Falkow, Curr. Opin. Microbiol. 1, 359 (1998). ~ . ~, e ~ ~ ~ • . ~ ~ . -o~ . ~ . o~ ~ O o~=~ ~ . ~ ,- ~ ~ ~ . ~ . ~ . ~ ~ ~ ' ~ : ~ . ~ . ~ ~ ~ ~ ~ ~ o ~ . ~ ;~,~m m ~ ~ ~ ~ •'- o • ~ ~ 4= ~ ,_~ ;...

1 mM ZnC12, and 1 mM iodoacetamide in 13 × 100-mm glass tubes. Include a blank without cells. 1% sodium dodecyl sulfate and 50 /~1 chloroform, vortex, and incubate at 37 ° for 5 min to permeabilize cells. Place tubes on ice for 5 min to cool. 0) to each individual tube, agitate, and place in 37 ° water bath. Note time. 0, 1 M KH2PO4, and place the tube in an ice-water bath to stop the reaction. Note time. Measure OD550 and OD420for each sample, using an assay mixture without cells as a blank. 75 x OD55o)] x 1000 Units activity = time (min) × OD6oo X volume cells (ml) Acknowledgments We thank J.

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