By P. Michael Conn
Neuroendocrine Peptide method
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Inayama, S. Nakanishi, and S. Numa, Nature (London) 195, 202 (1982). 6 U. Gubler, P. Seeburg, B. J. Hoffman, L. P. Gage, and S. Udenfriend, Nature (London) 295, 206 (1982). 7 M. Comb, P. H. Seeburg, J* Adelman, L. Eiden, and E. Herbert, Nature (London) 295, 663 (1982). 8 H. Kakidani, Y. Furutani, H. Takahashi, M. Noda, Y. Morimoto, T. Hirose, M. Asai, S. Inayama, S. Nakanishi, and S. Numa, Nature CLondon) 298, 245 (1982). NEUROENDOCRINE PEPTIDE METHODOLOGY Copyright © 1989 by Academic Press, Inc.
The purpose of integrating exogenous genes into the genome of a host cell is to create an “ immortal” hybrid cell line in order to facilitate long-term studies on the same clone. At this stage in the development of the tech nique, however, it is not possible to directly control the gene copy num ber (“ dosage” ) of the introduced gene or its position in the host genome. Typically, a number of independent clones are generated which contain a range of gene copies. Insertion into one or more cellular chromosomes permits study of recombination and positional effects on eukaryotic gene expression.
N. Wilcox, C. E. Gee, and J. L. Roberts, this series, Vol. 124, p. 510. 4 J. F. McKelvy, J. E. Krause, and J. D. White, this series, Vol. 103, p. 511. 5 M. Dean, R. A. Levine, W. Ran, M. S. Kindy, G. E. Sonenshein, and J. Campisi, J. Biol. Chem. 261, 9161 (1986). 6 R. Bravo, M. Neuberg, J. Burckhardt, J. Almendral, R. Wallich, and R. ) 48, 251 (1987). NEUROENDOCRINE PEPTIDE METHODOLOGY Copyright © 1989 by Academic Press, Inc. All rights of reproduction in any form reserved. 38 I. GENE EXPRESSION AND in Situ HYBRIDIZATION side triphosphate, and newly synthesized RNA is then isolated.