Development of a protein microarray using sequence-specific DNA binding domain on DNA chip surface.
A protein microarray based on DNA microarray platform was developed to identify protein-protein interactions in vitro. The conventional DNA chip surface by 156-bp PCR product was prepared for a substrate of protein microarray. High-affinity sequence-specific DNA binding domain, GAL4 DNA binding domain, was introduced to the protein microarray as fusion partner of a target model protein, enhanced green fluorescent protein. The target protein was oriented immobilized directly on the DNA chip surface. Finally, monoclonal antibody of the target protein was used to identify the immobilized protein on the surface. This study shows that the conventional DNA chip can be used to make a protein microarray directly, and this novel protein microarray can be applicable as a tool for identifying protein-protein interactions.
Choi
Yoo Seong
YS
School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Republic of Korea.
Pack
Seung Pil
SP
Yoo
Young Je
YJ
D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D004247:DNA