Application of dna binding protein tags as means for defined co-immobilization in biosensors

Identificador:  TDX:2675

Handle: https://hdl.handle.net/20.500.11797/TDX2675

Autors:  Aktas, Gülsen Betül

Resum:
The objective of this doctoral thesis is to explore the use of DNA binding proteins in biosensors for signal generation and amplification. The specificity and high affinity that these proteins exhibit for their respective double stranded DNA (dsDNA) sequence was thus exploited in order to develop a protein immobilization technique that allows the controlled co-immobilization of several proteins. The proposed system is based on the utilization of these DNA binding proteins as fusion tags for the target proteins to be immobilized and the use of dsDNA as the template to direct the protein co-immobilization by specific positioning of the target DNA sequences recognized by the DNA binding proteins. The preparation and characterization of two novel universal enzyme-DNA binding protein conjugates is described and their use for signal generation and amplification in biosensors for the detection of nucleic acid and protein targets is demonstrated. In addition, the suitability of these DNA binding proteins to directly immobilize DNA molecules is also shown.