Selection of aptamers against KMP-11 using colloidal gold during the SELEX process.
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.
Moreno
Miguel
M
Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Alcalá, 28871 Alcalá de Henares, Madrid, Spain.
Rincón
Esther
E
Piñeiro
David
D
Fernández
Gerónimo
G
Domingo
Alberto
A
Jiménez-Ruíz
Antonio
A
Salinas
Matilde
M
González
Víctor M
VM
D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D017979:Gold Colloid; D015151:Immunoblotting; D018314:Leishmania infantum / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009838:Oligodeoxyribonucleotides / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D016054:DNA, Protozoan; D004277:DNA, Single-Stranded; D017979:Gold Colloid; D008562:Membrane Glycoproteins; D009838:Oligodeoxyribonucleotides; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C090009:kinetoplastid membrane protein 11, Leishmania donovani