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

The seriously acclaimed laboratory common for greater than 40 years, Methods in Enzymology is likely one of the so much hugely revered courses within the box of biochemistry. seeing that 1955, every 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 comprises a lot fabric nonetheless appropriate today--truly a necessary ebook for researchers in all fields of existence sciences

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

The significantly acclaimed laboratory regular for greater than 40 years, tools in Enzymology is likely one of the so much hugely revered courses within the box of biochemistry. in view that 1955, each one quantity has been eagerly awaited, often 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 an important booklet for researchers in all fields of existence sciences

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

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M. S l a u c h a n d T. J. Silhavy, Methods E n z y m o l 204, 213 (1991). ] ........ BSR pR5528 ~ I I pRS552 pRS591 ~i~- I lITlll kan pRS577 lac "ZYA A. f B Mcs orientation pRS550 EcoRl Smal*** BamH! s imm434 CJIndattP imm434clind" lacZYA =. . . . bla ASClac 'ZYA FIG. 1. Adapted from Simons et aL 3 (A) Schematic diagram of plasmid vectors for constructing lac fusions. All of the plasmids are based on a pBR322 backbone and carry a fragment of the 3' end of the tetA gene. In addition, all of the plasmids carry four copies of the rrnB transcriptional terminator (Tl4) upstream (with respect to the lac operon) of the multiple cloning site.

J. Silhavy, and G. M. Weinstock, J. Bacteriol. 162, 1092 (1985). " From E. Bremer, T. J. Silhavy, and G. M. Weinstock, Gene 71, 177 (1988). t From E. Bremer, T. J. Silhavy, J. M. Weisemann, and G. M. Weinstock, J. Bacteriol. 158, 1084 (1984). the Aplac Mu series of hybrid phage vectors. 2"18'19The first example takes advantage of A vectors carrying mutations conditionally defective for multiple replicative functions. These vectors can be propagated in a bacterial host strain containing appropriate suppressor mutations.

W. Davis, H. A. Kirkpatrick, M. A. Goeden, D. J. Rose, B. Mau, and Y. Shao, Science 277, 1453 (1997). tl R. K. Saiki, S. Scharf, F. Faloona, K. B. Mullis, G. T. Horn, H. A. Erlich, and N. Arnheim, Science 230, 1350 (1985). ~2R. F. Moreira and C. J. Noren, Biotechniques 19, 56, 58 (1995). under the control of the strong, inducible placUV5 promoter. Recombination of fusions with low activity onto these A vectors can therefore be detected easily against a background of dark blue plaques. The fact that the inserts in ARS45 and ARS74 are in the opposite orientation with respect to those in ARS88 and ARS91 is not relevant.

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