Chemistry Reference
In-Depth Information
7.5.1.1
Selectivity and Efficiency of Site-selective DNA Scission
In the oligonucleotide additives used, a monophosphate group is bound to either the
3
0
- or the 5
0
-end via a (CH
2
)
12
linker (Figure 7.3). When these additives are combined
with the substrate DNA, gap-structures are formed at a predetermined site in the
Figure 7.3 DNA substrates and oligonucleotide additives used for
site-selective scission of single-stranded DNA.
Figure 7.4 (a) Polyacrylamide gel electrophoresis patterns for the hydrolysis
of DNA
(S5)
(
32
P-labelled at the 5
0
-end) by Ce(
IV
)/EDTA complex in the presence
of various additive oligonucleotides at pH 7.0 and 37
8
C. Lane 1, control
without both additive DNAs and Ce(
IV
)/EDTA; lane 2, control in the absence of
the additives [only with Ce(
IV
)/EDTA]; lane 3, DNA
(L)
-H/H-DNA
(R)
with Ce(
IV
)/
EDTA; lane 4, DNA
(L)
-L
12
-P/H-DNA
(R)
with Ce(
IV
)/EDTA; lane 5, DNA
(L)
-H/P-
L
12
-DNA
(R)
with Ce(
IV
)/EDTA; lane 6, DNA
(L)
-L
12
-P/P-L
12
-DNA
(R)
with Ce(
IV
)/
EDTA; M, the markers (authentic oligonucleotides having 3
0
-OH termini).
Reaction conditions: [DNA
(S5)
] = 1.0
M
,
[NaCl] = 100m
M
, and [Ce(
IV
)/EDTA] = 1.0m
M
at pH 7.0 (7.5m
M
Hepes buffer)
and 37
8
C. (b) Magnified versions of the main part in lanes 4-6.
l
M
, [each of the additive DNAs] = 2.0
l
