Sequence-specific binding of DNA and RNA to immobilized Nickel ions

Branislav Nastasijevic, Nicole A. Becker, Susan E. Wurster, L James Maher III

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Immobilization of divalent Nickel cations provides a tool for affinity purification of proteins containing hexahistidine tags. During experiments to generate single-stranded DNA aptamers to immobilized proteins we inadvertently identified DNA sequences with affinity for Nickel-nitrilotriacetic acid (Ni 2+-NTA) magnetic beads. Analysis of these aptamers revealed that affinity for the Ni 2+-NTA support requires only single-stranded sequences with multiple adenosine residues. Bound nucleic acids can be eluted with imidazole. A single-stranded dA 20 affinity tag (but not other homopolymer sequences) is sufficient for immobilization of double-stranded DNA PCR products on Ni 2+-NTA magnetic beads. Addition of an rA 20 sequence to an RNA transcript allowed its affinity capture on Ni 2+-NTA magnetic beads, suggesting an approach for purification of poly(A) mRNA.

Original languageEnglish (US)
Pages (from-to)420-425
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume366
Issue number2
DOIs
StatePublished - Feb 8 2008

Fingerprint

Immobilized Nucleic Acids
Nickel
RNA
Ions
DNA
His-His-His-His-His-His
Immobilization
Purification
Nucleotide Aptamers
Immobilized Proteins
Single-Stranded DNA
DNA sequences
Divalent Cations
Homopolymerization
Adenosine
Nucleic Acids
Cations
nickel nitrilotriacetic acid
Polymerase Chain Reaction
Messenger RNA

Keywords

  • Adenine
  • Aptamer
  • IMAC
  • In vitro selection
  • Nickel
  • Paramagnetic beads

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Sequence-specific binding of DNA and RNA to immobilized Nickel ions. / Nastasijevic, Branislav; Becker, Nicole A.; Wurster, Susan E.; Maher III, L James.

In: Biochemical and Biophysical Research Communications, Vol. 366, No. 2, 08.02.2008, p. 420-425.

Research output: Contribution to journalArticle

Nastasijevic, Branislav ; Becker, Nicole A. ; Wurster, Susan E. ; Maher III, L James. / Sequence-specific binding of DNA and RNA to immobilized Nickel ions. In: Biochemical and Biophysical Research Communications. 2008 ; Vol. 366, No. 2. pp. 420-425.
@article{dc3eff01897447938e0c03f53f817ab3,
title = "Sequence-specific binding of DNA and RNA to immobilized Nickel ions",
abstract = "Immobilization of divalent Nickel cations provides a tool for affinity purification of proteins containing hexahistidine tags. During experiments to generate single-stranded DNA aptamers to immobilized proteins we inadvertently identified DNA sequences with affinity for Nickel-nitrilotriacetic acid (Ni 2+-NTA) magnetic beads. Analysis of these aptamers revealed that affinity for the Ni 2+-NTA support requires only single-stranded sequences with multiple adenosine residues. Bound nucleic acids can be eluted with imidazole. A single-stranded dA 20 affinity tag (but not other homopolymer sequences) is sufficient for immobilization of double-stranded DNA PCR products on Ni 2+-NTA magnetic beads. Addition of an rA 20 sequence to an RNA transcript allowed its affinity capture on Ni 2+-NTA magnetic beads, suggesting an approach for purification of poly(A) mRNA.",
keywords = "Adenine, Aptamer, IMAC, In vitro selection, Nickel, Paramagnetic beads",
author = "Branislav Nastasijevic and Becker, {Nicole A.} and Wurster, {Susan E.} and {Maher III}, {L James}",
year = "2008",
month = "2",
day = "8",
doi = "10.1016/j.bbrc.2007.11.169",
language = "English (US)",
volume = "366",
pages = "420--425",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Sequence-specific binding of DNA and RNA to immobilized Nickel ions

AU - Nastasijevic, Branislav

AU - Becker, Nicole A.

AU - Wurster, Susan E.

AU - Maher III, L James

PY - 2008/2/8

Y1 - 2008/2/8

N2 - Immobilization of divalent Nickel cations provides a tool for affinity purification of proteins containing hexahistidine tags. During experiments to generate single-stranded DNA aptamers to immobilized proteins we inadvertently identified DNA sequences with affinity for Nickel-nitrilotriacetic acid (Ni 2+-NTA) magnetic beads. Analysis of these aptamers revealed that affinity for the Ni 2+-NTA support requires only single-stranded sequences with multiple adenosine residues. Bound nucleic acids can be eluted with imidazole. A single-stranded dA 20 affinity tag (but not other homopolymer sequences) is sufficient for immobilization of double-stranded DNA PCR products on Ni 2+-NTA magnetic beads. Addition of an rA 20 sequence to an RNA transcript allowed its affinity capture on Ni 2+-NTA magnetic beads, suggesting an approach for purification of poly(A) mRNA.

AB - Immobilization of divalent Nickel cations provides a tool for affinity purification of proteins containing hexahistidine tags. During experiments to generate single-stranded DNA aptamers to immobilized proteins we inadvertently identified DNA sequences with affinity for Nickel-nitrilotriacetic acid (Ni 2+-NTA) magnetic beads. Analysis of these aptamers revealed that affinity for the Ni 2+-NTA support requires only single-stranded sequences with multiple adenosine residues. Bound nucleic acids can be eluted with imidazole. A single-stranded dA 20 affinity tag (but not other homopolymer sequences) is sufficient for immobilization of double-stranded DNA PCR products on Ni 2+-NTA magnetic beads. Addition of an rA 20 sequence to an RNA transcript allowed its affinity capture on Ni 2+-NTA magnetic beads, suggesting an approach for purification of poly(A) mRNA.

KW - Adenine

KW - Aptamer

KW - IMAC

KW - In vitro selection

KW - Nickel

KW - Paramagnetic beads

UR - http://www.scopus.com/inward/record.url?scp=37448998768&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=37448998768&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2007.11.169

DO - 10.1016/j.bbrc.2007.11.169

M3 - Article

C2 - 18068116

AN - SCOPUS:37448998768

VL - 366

SP - 420

EP - 425

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 2

ER -