Oligonucleotide circularization by template-directed chemical ligation.
An efficient method for producing the covalent closure of oligonucleotides on complementary templates by the action of BrCN was developed. A rational design of linear precursor oligonucleotides was studied, and the effect of factors such as oligonucleotide concentration and oligomer-template length ratio was evaluated. The efficiency of circularization was shown to correlate well with the secondary structure of the precursor oligomer (as predicted by a simple computer analysis), hairpin-like structures bearing free termini clearly favouring the circularization reaction. A novel idea, consisting of the incorporation of non-nucleotide insertions in the precursor oligomer (namely, 1,2-dideoxy-D-ribofuranose residues), may render this method universal and highly effective. An original set of assays was developed to confirm the circular structure of the covalently closed oligonucleotides.
Dolinnaya
N G
NG
GENSET, Paris, France.
Blumenfeld
M
M
Merenkova
I N
IN
Oretskaya
T S
TS
Krynetskaya
N F
NF
Ivanovskaya
M G
MG
Vasseur
M
M
Shabarova
Z A
ZA
D001483:Base Sequence; D003488:Cyanogen Bromide / Q000737:chemistry; D004270:DNA, Circular / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry; D006860:Hydrogen Bonding; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry*; D013698:Templates, Genetic
D004270:DNA, Circular; D004277:DNA, Single-Stranded; D009841:Oligonucleotides; D003488:Cyanogen Bromide