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Open image in new window Figure 2 DraIII digests star site sequence in asymmetrical pattern.
First, CATNNNGTG was identified as a strong star site for DraIII.
A larger 22.6-kb plasmid DNA pXba was used to verify the predicted star site.
(B) DraIII star site in pUC19 was predicted to be the CATGTTGTG site.
(C and D) DraIII digests a star site sequence in an asymmetrical manner.
pXba contains three DraIII canonical sites (nt 5816, 16551 and 17404 respectively) and one predicted star site CATGTTGTG (nt 21910).
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It is difficult to map the DraIII star sites in λ DNA because of its large size (48.5-kb) and the large number of possible star sites.
A smaller 2.7-kb pUC19 DNA was used to determine DraIII star sites.
Open image in new window Figure 1 Determination ofDraIII Fidelity Index FII) and star sites.
Because star activity is relatively weak, the star sites of only a few enzymes have been defined.
According to this hypothesis, REases showing high star activity, such as DraIII, may have a lower thermodynamic barrier to adopt the cleavage competent conformation with some star sites.
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