Self-assembling protein arrays on DNA chips by auto-labeling fusion proteins with a single DNA address.
The high-throughput deposition of recombinant proteins on chips, beads or biosensor devices would be greatly facilitated by the implementation of self-assembly concepts. DNA-directed immobilization via conjugation of proteins to an oligonucleotide would be preeminently suited for this purpose. Here, we present a unique method to attach a single DNA address to proteins in one step during the purification from the E. coli lysate by fusion to human O6-alkylguanine-DNA-alkyltransferase (SNAP-tag) and the Avitag. Use of the conjugates in converting a DNA chip into a protein chip by self assembly is demonstrated.
Jongsma
Maarten A
MA
Plant Research International, Wageningen, The Netherlands. maarten.jongsma@wur.nl
Litjens
Ralph H G M
RH
D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D005240:Feasibility Studies; D015335:Molecular Probes / Q000737:chemistry*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000737:chemistry; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D050766:Qb-SNARE Proteins / Q000737:chemistry; D050767:Qc-SNARE Proteins / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000737:chemistry*
D015335:Molecular Probes; D050766:Qb-SNARE Proteins; D050767:Qc-SNARE Proteins; D011993:Recombinant Fusion Proteins; C100493:SNAP23 protein, human; D019853:O(6)-Methylguanine-DNA Methyltransferase