Identification of cis-regulatory sequences in ascidian ribosomal DNA using a rapid filter binding assay.

Degnan, B. M. and Lavin, M. F. (1991) Identification of cis-regulatory sequences in ascidian ribosomal DNA using a rapid filter binding assay.. Gene, 109 2: 249-253. doi:10.1016/0378-1119(91)90616-J


Author Degnan, B. M.
Lavin, M. F.
Title Identification of cis-regulatory sequences in ascidian ribosomal DNA using a rapid filter binding assay.
Journal name Gene   Check publisher's open access policy
ISSN 0378-1119
Publication date 1991-12-30
Sub-type Other
DOI 10.1016/0378-1119(91)90616-J
Volume 109
Issue 2
Start page 249
End page 253
Total pages 5
Place of publication Amsterdam
Publisher Elsevier B.V.
Language eng
Subject 270504 Invertebrate Biology
270200 Genetics
Abstract Using 32P-labelled random primed ribosomal DNA (rDNA) from the ascidian, Herdmania momus, multiple and large-scale filter-binding assays were performed to identify cis-regulatory sequences interacting with H. momus oocyte germinal vesicle protein. A vacublot apparatus was used to isolate DNA-protein complexes, providing a means of filtering multiple binding reactions simultaneously and for isolating sufficient amounts of bound DNA for further investigations. DNA bound to the filter was used to identify unknown cis-elements in the rDNA by Southern-blot analysis. The trapped rDNA hybridized specifically to the intergenic spacer, a region which contains cis-regulatory sequences that interact with rDNA transcription factors in several other species. Gel shift analysis of intergenic spacer fragments and native Southwestern blots confirmed that cis-elements were localized in the rDNA intergenic specer. In principle, this allows for the rapid identification of cis-fegulatory sequences within any large, cloned DNA fragment which interact with nuclear extract.
Keyword Herdmania momus
intergenic spacer
marine invertebrates
tunicates
transcription factor
promoter
vacublot recombinant DNA
Q-Index Code CX
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Other
Collections: Degnan Laboratory
School of Biological Sciences Publications
 
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Created: Thu, 06 Dec 2007, 02:17:35 EST by Laura McTaggart