|
|
|
|
|
|
|
||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | ||||
Development, Vol 113, Issue 2 385-398, Copyright © 1991 by Company of Biologists
JOURNAL ARTICLES |
DL Livant, BR Hough-Evans, JG Moore, RJ Britten and EH Davidson
Division of Biology, California Institute of Technology, Pasadena 91125.
The object of these experiments was to determine whether competitive titration in vivo of factors required for expression of the CyIIIa.CAT fusion gene would affect expression of the endogenous CyIIIa gene in the same embryos. Earlier work showed that expression of this fusion gene after injection into sea urchin eggs is stoichiometrically reduced when low molar excesses of DNA fragments containing only its regulatory domain are coinjected. In order to compare endogenous (i.e. CyIIIa) and exogenous (i.e. CyIIIa.CAT) expression simultaneously in embryos bearing excess competitor regulatory DNA, we developed, and here describe, a new procedure for generating transgenic sea urchin embryos in which all of the cells in many embryos, and most in others, bear the exogenous DNA. Such large reduction of mosaicism can be achieved by multiple injection of the exogenous DNA fragments into fertilized eggs. Using this method, we demonstrate that at a level of competitor DNA incorporation which reduces CyIIIa.CAT expression by 85%, endogenous CyIIIa mRNA levels are wholly unaffected. Nor is spatial expression of the endogenous CyIIIa gene disturbed. Since the CyIIIa.CAT genes are properly expressed under control of the CyIIIa regulatory sequences, they must participate in the same set of necessary DNA-protein interactions. However, we infer from the results that we report here that the regulatory complexes in the endogenous CyIIIa gene are greatly stabilized relative to those of the exogenous CyIIIa.CAT genes.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati 
Twitter What's this?
This article has been cited by other articles:
|
|
 |
|
  J. Smith, C. Theodoris, and E. H. Davidson A Gene Regulatory Network Subcircuit Drives a Dynamic Pattern of Gene Expression Science, November 2, 2007; 318(5851): 794 - 797. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. J. Dmochowski, J. E. Dmochowski, P. Oliveri, E. H. Davidson, and S. E. Fraser Quantitative imaging of cis-regulatory reporters in living embryos PNAS, October 1, 2002; 99(20): 12895 - 12900. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Li, A. Streit, B. Robertson, and W. B. Wood Evidence for Multiple Promoter Elements Orchestrating Male-Specific Regulation of the her-1 Gene in Caenorhabditis elegans Genetics, May 1, 1999; 152(1): 237 - 248. [Abstract] [Full Text]
|
|
|
|
|
|
L. D. Bogarad, M. I. Arnone, C. Chang, and E. H. Davidson Interference with gene regulation in living sea urchin embryos: Transcription factor Knock Out (TKO), a genetically controlled vector for blockade of specific transcription factors PNAS, December 8, 1998; 95(25): 14827 - 14832. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Arnone, L. Bogarad, A Collazo, C. Kirchhamer, R. Cameron, J. Rast, A Gregorians, and E. Davidson Green Fluorescent Protein in the sea urchin: new experimental approaches to transcriptional regulatory analysis in embryos and larvae Development, January 11, 1997; 124(22): 4649 - 4659. [Abstract] [PDF]
|
|
|
|
|
|
C. Yuh and E. Davidson Modular cis-regulatory organization of Endo16, a gut-specific gene of the sea urchin embryo Development, January 4, 1996; 122(4): 1069 - 1082. [Abstract] [PDF]
|
|
|
|
|
|
C. Kirchhamer and E. Davidson Spatial and temporal information processing in the sea urchin embryo: modular and intramodular organization of the CyIIIa gene cis-regulatory system Development, January 1, 1996; 122(1): 333 - 348. [Abstract] [PDF]
|
|
|
|
|
|
K. Makabe, C. Kirchhamer, R. Britten, and E. Davidson Cis-regulatory control of the SM50 gene, an early marker of skeletogenic lineage specification in the sea urchin embryo Development, January 7, 1995; 121(7): 1957 - 1970. [Abstract] [PDF]
|
|
|
|
|
|
D. Wang, C. Kirchhamer, R. Britten, and E. Davidson SpZ12-1, a negative regulator required for spatial control of the territory-specific CyIIIa gene in the sea urchin embryo Development, January 4, 1995; 121(4): 1111 - 1122. [Abstract] [PDF]
|
|
