Applications of Chimeric Genes and Hybrid Proteins Part A: by Melvin I. Simon, Jeremy Thorner, Scott D. Emr, John N. PDF

By Melvin I. Simon, Jeremy Thorner, Scott D. Emr, John N. Abelson

ISBN-10: 0121822273

ISBN-13: 9780121822279

The severely acclaimed laboratory usual for greater than 40 years, Methods in Enzymology is without doubt one of the such a lot hugely revered guides within the box of biochemistry. given that 1955, each one quantity has been eagerly awaited, often consulted, and praised through researchers and reviewers alike. Now with greater than three hundred volumes (all of them nonetheless in print), the sequence includes a lot fabric nonetheless suitable today--truly an important book for researchers in all fields of existence sciences

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Lysates produced in this way contain a heterogeneous population of [2] CONSTRUCTING lac FUSIONS IN E. coli 29 phage particles. The desired specialized transducing phage particles can be selected by following the procedure described previously for targeting lac fusions to the chromosome. The recipient strain should be rec +. Lysogens should be tested by cross-streaking as described, with AcI and Avir as test phage. Most lysogens selected on minimal lactose will have lost the Mu sequences from the opposite side of the original integrated prophage.

In comparison to these newer technologies, flgalactosidase, with its lengthy history (relative to the age of molecular genetics), seems almost old-fashioned. However, it is precisely this extensive history that makes the Lac system so useful. In the hands of an experienced investigator, exquisitely detailed questions can still be addressed with little more than agar plates and toothpicks. It seems likely, therefore, that Lac will remain an often-used weapon in the arsenal of the bacterial geneticist for the foreseeable future.

Conversely, screening pools of random LacZ chromosomal insertions can identify targets of a regulator (either transcriptional or translational). Finally, fl-galactosidase remains useful in the study of translational regulation, although certain caveats must be considered in studying LacZ protein fusions. Rather than revisit techniques that are no longer of significant interest, we will discuss a more limited selection of those applications for which /3-galactosidase is generally most useful. We will also present a limited set of up-to-date protocols for making specific or random lac fusions (both transcriptional and translational).

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Applications of Chimeric Genes and Hybrid Proteins Part A: Gene Expression and Protein Purification by Melvin I. Simon, Jeremy Thorner, Scott D. Emr, John N. Abelson


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