Target Detection Assay (TDA): a versatile procedure to determine DNA blinding sites as demonstrated on SP1 protein (original) (raw)
Journal Article
,
Basel Institute for Immunology, Grenzacherstraβe 487
CH-4005 Basel, Switzerland
Search for other works by this author on:
Basel Institute for Immunology, Grenzacherstraβe 487
CH-4005 Basel, Switzerland
Search for other works by this author on:
PDF Cite
Hans-jürgen Thiesen, Christian Bach, Target Detection Assay (TDA): a versatile procedure to determine DNA blinding sites as demonstrated on SP1 protein, Nucleic Acids Research, Volume 18, Issue 11, 11 June 1990, Pages 3203–3209, https://doi.org/10.1093/nar/18.11.3203
Close
Navbar Search Filter Mobile Enter search term Search
Abstract
We developed a rapid method designated TargetDetection Assay (TDA) to determine DNA binding sites for putative DNA binding proteins. A purified, functionally active DNA binding protein and a pool of random double-stranded oligonucleotides harbouring PCR primr sites at each end are included the TDA cycle which consists of four separate steps: a DNA protein incubation step, a protein DNA complex separation step , a DNA elutfon step and a polymerase chain reaction (PCR) DNA amplification step. The stringency of selection can be increased in consecutive TDA cycles. Since tiny amounts of retained DNA can be rescued by PCR, buffer systems, salt concentrations and competitor DNA contents can be varied In order to determine high affinity binding sites for the protein of choice. To test the efficiency of the TDA procedure potential DNA binding sites were selected by the DNA binding protein SP1 from a pool of oligonucleotides with random nucleotides at 12 positions. Target sites selected by recomblnant SP1 closely matched the SP1 consensus site. If DNA recognition sites have to be determined for known, mutated or putative DNA binding proteins, the Target Detection Assay (TDA) is a versatile and rapid technique for consideration.
This content is only available as a PDF.
© 1990 Oxford University Press
I agree to the terms and conditions. You must accept the terms and conditions.
Submit a comment
Name
Affiliations
Comment title
Comment
You have entered an invalid code
Thank you for submitting a comment on this article. Your comment will be reviewed and published at the journal's discretion. Please check for further notifications by email.
Citations
Views
Altmetric
Metrics
Total Views 176
25 Pageviews
151 PDF Downloads
Since 1/1/2017
| Month: | Total Views: |
|---|---|
| January 2017 | 3 |
| March 2017 | 1 |
| April 2017 | 1 |
| July 2017 | 3 |
| August 2017 | 3 |
| October 2017 | 2 |
| November 2017 | 1 |
| December 2017 | 11 |
| January 2018 | 17 |
| February 2018 | 9 |
| March 2018 | 10 |
| April 2018 | 13 |
| May 2018 | 1 |
| July 2018 | 2 |
| February 2019 | 1 |
| May 2019 | 1 |
| November 2019 | 1 |
| December 2019 | 1 |
| February 2020 | 1 |
| September 2020 | 1 |
| December 2020 | 1 |
| January 2021 | 1 |
| August 2021 | 1 |
| September 2021 | 1 |
| November 2021 | 1 |
| January 2022 | 1 |
| February 2022 | 2 |
| April 2022 | 1 |
| June 2022 | 1 |
| August 2022 | 2 |
| September 2022 | 7 |
| October 2022 | 3 |
| November 2022 | 3 |
| December 2022 | 3 |
| January 2023 | 1 |
| February 2023 | 3 |
| March 2023 | 1 |
| May 2023 | 1 |
| July 2023 | 4 |
| August 2023 | 3 |
| October 2023 | 2 |
| November 2023 | 2 |
| December 2023 | 2 |
| January 2024 | 3 |
| February 2024 | 3 |
| March 2024 | 6 |
| April 2024 | 6 |
| May 2024 | 8 |
| June 2024 | 5 |
| July 2024 | 4 |
| August 2024 | 5 |
| September 2024 | 4 |
| October 2024 | 2 |
Citations
202 Web of Science
×
Email alerts
Citing articles via
More from Oxford Academic