Download e-book for iPad: Applications of Chimeric Genes and Hybrid Proteins Part A: by Melvin I. Simon, Jeremy Thorner, Scott D. Emr, John N.

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

ISBN-10: 0121822273

ISBN-13: 9780121822279

The seriously acclaimed laboratory commonplace 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. considering the fact that 1955, each one quantity has been eagerly awaited, often consulted, and praised via researchers and reviewers alike. Now with greater than three hundred volumes (all of them nonetheless in print), the sequence comprises a lot fabric nonetheless appropriate today--truly a vital ebook for researchers in all fields of lifestyles sciences

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Additional info for Applications of Chimeric Genes and Hybrid Proteins Part A: Gene Expression and Protein Purification

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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. In comparison to these newer technologies, flgalactosidase, with its lengthy history (relative to the age of molecular genetics), seems almost old-fashioned.

This is accomplished by using UV irradiation to stimulate excision of the A prophage by illegitimate recombination. (D) Illegitimate recombination releases the h prophage and some of the DNA flanking it. If transmission of the fusion is selected (by selecting for lac activity, for example), the sequence upstream of the lac operon can mediate transfer of the fusion to the chromosome by homologous recombination. Because transmission of the sequence to the left is under selection, it is less likely that additional DNA to the right of the prophage will also be packaged into the h phage particle.

Walsh-Weller,Methods Mol. Biol. 98, 121 (1998). 18 GENE FUSIONS [21 prefer to use combinations that produce lysogens in which the fusion is linked to a drug resistance marker. This facilitates the movement of the fusions from one strain to another by generalized transduction. Paradoxically, marked fusions can complicate strain construction because of the loss of the ability to use the marker on the A phage to move other mutations or maintain plasmids. For illustrative purposes, we will consider a case in which the lysogen is not marked.

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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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