By Melvin I. Simon, Jeremy Thorner, Scott D. Emr, John N. Abelson
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Additional info for Applications of Chimeric Genes and Hybrid Proteins Part A: Gene Expression and Protein Purification
Vortex the tubes briefly and then centrifuge at 4500g for 10 min. 9. Transfer the supernatant to sterile screw-cap tubes (taking care to avoid transferring any of the chloroform) and store in the dark at 4°. Production of Single-Copy Lac Fusion ~t Lysogens Single-copy Lac fusion-containing lysogens may be conveniently placed at two different sites in the bacterial chromosome. The most common choice of sites is at the wild-type ~ attachment site (att) located at 17' on  CONSTRUCTING lac FUSIONS IN g.
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.
METHODS IN ENZYMOLOGY, VOL. 326 Copyright © 2000 by Academic Press All rights of reproduction in any form reserved. 00 12 GENE FUSIONS [21 article m a y be found elsewhere. 4'6-8 In addition, we will attempt to preserve some of the " L a c Z lore" that is disappearing with the increased use of other protein tags and reporter systems. In particular, we will discuss the use of indicator and selector media. P r o d u c t i o n of LacZ F u s i o n s T h r e e separate partially overlapping nomenclatures exist to describe lac fusions.
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