Efficiency of T4 DNA ligase-catalyzed end joining after S1 endonuclease treatment on duplex DNA containing single-stranded portions.
Biochim Biophys Acta
; 656(1): 123-7, 1981 Nov 27.
Article
in En
| MEDLINE
| ID: mdl-6171303
ABSTRACT
Covalently closed-circular, superhelical SV4O DNA was used in all experiments. EcoRI endonuclease- and HpaII endonuclease-generated unit-length linear duplex DNAs were digested with S1 endonuclease under the conditions where single-stranded CNA was completely converted into the acid-soluble form. These were subjected to an end-to-end joining test with T4 DNA ligase. The ligation efficiency was significantly lower than that of the flush-ended linear duplex DNAs which were generated by both HpaI endonuclease digestion and the matching up of EcoRI-generated sticky end with Escherichia coli DNA polymerase I (Klenow fraction). However, the ligation efficiency of the S1-treated DNAs increased up to same level as the flush-ended DNA upon treatment with E. coli DNA polymerase I . Similar results were obtained in the case of S1 -generated unit-length linear duplex DNA. S1 does cleave both strands of superhelical DNA at unbasepaired sites.
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Database:
MEDLINE
Main subject:
DNA Ligases
/
Polynucleotide Ligases
/
DNA, Superhelical
/
DNA, Viral
/
Deoxyribonucleases, Type II Site-Specific
/
Endonucleases
Language:
En
Journal:
Biochim Biophys Acta
Year:
1981
Type:
Article