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
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Base Sequence
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DNA, Protozoan / genetics
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DNA, Protozoan / isolation & purification*
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DNA, Protozoan / metabolism
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DNA, Single-Stranded / genetics
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DNA, Single-Stranded / isolation & purification
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DNA, Single-Stranded / metabolism
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Enzyme-Linked Immunosorbent Assay
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Gold Colloid
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Immunoblotting
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Leishmania infantum / genetics*
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Membrane Glycoproteins / genetics*
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Membrane Glycoproteins / metabolism
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Nucleic Acid Amplification Techniques / methods*
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Oligodeoxyribonucleotides / genetics
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Oligodeoxyribonucleotides / isolation & purification
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Oligodeoxyribonucleotides / metabolism
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Protein Binding
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Protozoan Proteins / genetics*
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Protozoan Proteins / metabolism
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
Substances
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DNA, Protozoan
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DNA, Single-Stranded
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Gold Colloid
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Membrane Glycoproteins
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Oligodeoxyribonucleotides
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Protozoan Proteins
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Recombinant Proteins
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kinetoplastid membrane protein 11, Leishmania donovani