Selection of aptamers against KMP-11 using colloidal gold during the SELEX process

Biochem Biophys Res Commun. 2003 Aug 22;308(2):214-8. doi: 10.1016/s0006-291x(03)01352-4.

Abstract

SELEX procedure is a methodology in which single stranded oligonucleotides are selected from a wide variety of sequences based on their interaction with a target molecule. We have designed a novel SELEX methodology using colloidal gold to select high affinity single stranded DNA aptamers against Leishmania infantum KMP-11. Kinetoplastid membrane protein-11 (KMP-11) is a major component of the cell membrane of kinetoplastid parasites. Although its function is not known, the fact that KMP-11 is a cytoskeleton-associated protein suggests that it may be involved in mobility or in some other aspects of the flagellar structure. We have isolated a single stranded DNA aptamer population that binds specifically to L. infantum KMP-11. This population has been characterized in a series of in vitro experiments suggesting that it may be used as a powerful tool to further investigate the role of KMP-11 during Leishmania development and/or as a diagnostic tool in Leishmania infection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA, Protozoan / genetics
  • DNA, Protozoan / isolation & purification*
  • DNA, Protozoan / metabolism
  • DNA, Single-Stranded / genetics
  • DNA, Single-Stranded / isolation & purification
  • DNA, Single-Stranded / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Gold Colloid
  • Immunoblotting
  • Leishmania infantum / genetics*
  • Membrane Glycoproteins / genetics*
  • Membrane Glycoproteins / metabolism
  • Nucleic Acid Amplification Techniques / methods*
  • Oligodeoxyribonucleotides / genetics
  • Oligodeoxyribonucleotides / isolation & purification
  • Oligodeoxyribonucleotides / metabolism
  • Protein Binding
  • Protozoan Proteins / genetics*
  • Protozoan Proteins / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • DNA, Protozoan
  • DNA, Single-Stranded
  • Gold Colloid
  • Membrane Glycoproteins
  • Oligodeoxyribonucleotides
  • Protozoan Proteins
  • Recombinant Proteins
  • kinetoplastid membrane protein 11, Leishmania donovani