| dc.contributor |
David G . Gorenstein |
|
| dc.contributor |
Bruce A . Luxon |
|
| dc.contributor |
William D . Willis |
|
| dc.contributor |
L .Lilian Chan |
|
| dc.creator |
Zhao , Xu |
|
| dc.date |
2008 -06 -17 |
|
| dc.date.accessioned |
2011 -04 -27T21 :37 :54Z |
|
| dc.date.available |
2011 -04 -27T21 :37 :54Z |
|
| dc.date.issued |
2011 -04 -27T21 :37 :54Z |
|
| dc.identifier |
http : / /etd .utmb .edu /theses /available /etd -07252007 -082849 / |
|
| dc.identifier.uri |
http : / /hdl .handle .net /2249 .1 /35722 |
|
| dc.description |
Nucleic acid research has expanded the way we can intervene with biological systems . Especially , oligonucleotide agents (ODN or aptamers ) are believed to affect cell function via complementary recognition or binding to specific proteins by forming tertiary structure . This opens new ways in therapeutics and diagnostics . The phosphoro - mono - /di - thioate substitutions in the backbone (termed thioaptamer ) grants ODN nuclease resistance and higher binding affinity .
A bead -based combinatorial library , in which every bead contains a unique species of aptamers , provides a promising platform for selection of aptamers and thioaptamers . To successfully screen the bead -based library , 2D layout of beads in gel and on bead screening model is proposed . To develop the 2D layout of beads and its corresponding functional assays , a model system is first established : NF -kappa B proteins were expressed , purified and characterized . Thioaptamer XBY6 , which specifically targets NF -kappa B protein , and its natural origin , I -kappa B were synthesized and verified . Thioaptamer purification using FPLC and HPLC was also investigated , and several 5 -funtionalized thioaptamers were successfully purified . Electrophoretic mobility shifting assay (EMSA ) has been used to verify XBY6 binding , and ELISA assay has been used to verify I -kappa B binding towards human recombinant NF -kappa B protein . Preliminary study of bead in 2D gel showed applicability of bead -based selection and thus on bead functional assays were developed . Both double strand one species library with I -kappa B sequence and a 212=4096 different species beads library were constructed and verified . The library was then tested using on -bead EMSA like assay and ELISA assay . Both assays showed encouraging results for 2D layout selection and further enhancement of visualization (signal /noise improvement ) is discussed .
The project suggested that 2D layout of beads in gel (PAGE ) is well suited for parallel high -throughput selection of thioaptamers and aptamers , thus paving a new way for drug discovery and future therapeutics and diagnostics . |
|
| dc.format |
application /pdf |
|
| dc.language |
en |
|
| dc.publisher |
UTMB |
|
| dc.rights |
restricted |
|
| dc.rights |
I hereby certify that , if appropriate , I have obtained and attached hereto a written permission statement from the owner (s ) of each third party copyrighted matter to be included in my thesis , dissertation , or project report , allowing distribution as specified below . I certify that the version I submitted is the same as that approved by my advisory committee . I hereby grant to University of Texas Medical Branch or its agents the non -exclusive license to archive and make accessible , under the conditions specified below , my thesis , dissertation , or project report in whole or in part in all forms of media , now or hereafter known . I retain all other ownership rights to the copyright of the thesis , dissertation or project report . I also retain the right to use in future works (such as articles or books ) all or part of this thesis , dissertation , or project report . |
|
| dc.source |
http : / /etd .utmb .edu /theses /available /etd -07252007 -082849 / |
|
| dc.subject |
Human Biological Chemistry and Genetics |
|
| dc.title |
2D Layout of Bead Based Thioaptamer /Aptamer Selection Platform for Therapeutics and Diagnostics |
|
| dc.type |
text |
|