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By Goeddel D.V., Emr S.D., Gold L. (eds.)

The tools offered during this quantity will permit the reader to layout powerful innovations for the expression of cloned genes and cDNAs and should turn out important in fixing the vast majority of expression difficulties one is probably going to come across.

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12, 3937 (1984). 66 B. E. Schoner, R. M. Belagaje, and R. G. Schoner, Proc. Natl. Acad. Sci. A. 83, 8506 (1986). 67 H. A. de Boer and R. A. Kastelein, in "Maximizing Gene Expression" (W. Reznikoffand L. ), p. 225. Butterworth, Stoneham, Massachusetts, 1986. 6s M. Bulmer, Nature (London) 325, 728 (1987). 69 M. Robinson, R. Lilley, S. Little, J. S. Emtage, G. Yarranton, P. Stephens, A. Millican, M. Eaton, and G. Hymphreys, Nucleic Acids Res. 12, 6663 (1984). 70 L. Holm, Nucleic Acids Res. 14, 3075 (1986).

196, 85 (1987). 31 j. D. Gralla, A. J. Carpousis, and J. E. Stefano, Biochemistry 19, 5864. 32 A. J. Carpousis, J. E. Stefano, and J. D. Gralla, J. Mol. Biol. 157, 619 (1982). 33 A. J. Carpousis and J. D. Gralla, J. Mol. Biol. 183, 165 (1985). D. E. Johnston and W. R. MeClure, in "RNA Polymerase" (R. Losick and M. ), p. 413. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1976. [4] E. coli PROMOTER FUNCTION 43 within this region? 5 In another case, it was shown that a transcript starting with guanosine produced fewer cycling products than a neady identical transcript initiated with adenosine, a2 Cycling reactions are likely ended when tr is released, contacts with the promoter are broken, and a downstream elongation complex is formed.

Mol. Biol. 195, 949 (1987). 60 y. Flashner and J. Gralla, Proc. Natl. Acad. Sci. A. 85, 8968 (1988). 6t M. Mossing and T. Record, Science 233, 889 (1986). 62 H. Kramer, M. Amouyal, A. Nordheim, and B. M011er-Hill, EMBO J. 7, 547 (1988). 50 EXPRESSION IN g . coli [4] tive internal operator and thus are probably not fully repressible. Therefore, for tightest repression, one probably wants a tight DNA-binding protein that is capable of forming tight higher order structures when interacting with a suitably spaced operator element of optimal sequence.

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