Cameron, R. Andrew and Oliveri, Paola and Wyllie, Jane and Davidson, Eric H. (2004) cis-Regulatory activity of randomly chosen genomic fragments from the sea urchin. Gene Expression Patterns, 4 (2). pp. 205-213. ISSN 1567-133X http://resolver.caltech.edu/CaltechAUTHORS:20121024-164516716
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In order to determine the frequency and variety of cis-regulatory elements that function during embryonic development of Strongylocentrotus purpuratus, we constructed a GFP expression vector in which to test the activity of randomly chosen genomic DNA fragments that includes a promiscuous basal promoter from the endo16 gene. This vector was demonstrated to serve as a cis-regulatory element trap. We used it to carry out an initial test for the occurrence of elements that would promote GFP expression in this genome. In the screen reported here 108 different randomly chosen DNA fragments (av. 3.8 kb) were inserted in the vector, and each was injected into >200 zygotes. Surprisingly, 13% of the fragments tested yielded detectable levels of GFP expression in the recipient embryos. Specific patterns observed included expression in endoderm, in aboral ectoderm, and in pigment cells. The majority of active constructs expressed GFP in all spatial domains of the embryo. Elements with detectable cis-regulatory activity in the embryo occur in the sample screened, on the average, about every 30 kb, and the genome must include many thousands of such elements. On further analysis one isolate was shown to contain a gut specific element as well as one that controls expression in the secondary mesenchyme cells.
|Additional Information:||© 2003 Elsevier B.V. Received 6 June 2003; Revised 28 August 2003; Accepted 28 August 2003; Available online 4 November 2003. We thank Dr James Wallace, University of Washington and Dr David Mathog, Division of Biology, California Institute of Technology, for aid with computer searches. Dr M. Ina Arnone generated the EpGFP vector that served as the parent plasmid from which our vector was constructed. We thank Jonathan Rast for thoughtful comments on the manuscript. This work was supported by grants from NIH RR 06591 and Human Frontier Science Program RG0790. These authors contributed equally to this work.|
|Subject Keywords:||cis-Regulatory element; Transcription regulation; Gene transfer; Sea urchin|
|Official Citation:||R.Andrew Cameron, Paola Oliveri, Jane Wyllie, Eric H. Davidson, cis-Regulatory activity of randomly chosen genomic fragments from the sea urchin, Gene Expression Patterns, Volume 4, Issue 2, March 2004, Pages 205-213, ISSN 1567-133X, 10.1016/j.modgep.2003.08.007. (http://www.sciencedirect.com/science/article/pii/S1567133X0300173X)|
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|Deposited By:||Aucoeur Ngo|
|Deposited On:||25 Oct 2012 14:45|
|Last Modified:||25 Oct 2012 14:45|
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