{"title":"S1内切酶处理含单链部分的双链DNA后，T4 DNA连接酶催化末端连接的效率","authors":"Kazuo Shishido, Tadahiko Ando","doi":"10.1016/0005-2787(81)90035-6","DOIUrl":null,"url":null,"abstract":"<div><p>Covalently closed-circular, superhelical SV40 DNA was used in all experiments. <em>Eco</em>RI endonuclease- and <em>Hpa</em>II enonuclease-generated unit-length linear duplex DNAs were digested with S<sub>1</sub> endonuclease under the conditions where single-stranded DNA 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 <em>Hpa</em>I endonuclease digestion and the matching up of <em>Eco</em>RI-generated sticky end with <em>Escherichia coli</em> DNA polymerase I (Klenow fraction). However, the ligation efficiency of the S<sub>1</sub>-treated DNAs increased up to same level as the flush-ended DNA upon treatment with <em>E. coli</em> DNA polymerase I. Similar results were obtained in the case of S<sub>1</sub>-generated unit-length linear duplex DNA. S<sub>1</sub> does cleave both strands of superhelical DNA at unbasepaired sites.</p></div>","PeriodicalId":100164,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","volume":"656 1","pages":"Pages 123-127"},"PeriodicalIF":0.0000,"publicationDate":"1981-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0005-2787(81)90035-6","citationCount":"10","resultStr":"{\"title\":\"Efficiency of T4 DNA ligase-catalyzed end joining after S1 endonuclease treatment on duplex DNA containing single-stranded portions\",\"authors\":\"Kazuo Shishido, Tadahiko Ando\",\"doi\":\"10.1016/0005-2787(81)90035-6\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Covalently closed-circular, superhelical SV40 DNA was used in all experiments. <em>Eco</em>RI endonuclease- and <em>Hpa</em>II enonuclease-generated unit-length linear duplex DNAs were digested with S<sub>1</sub> endonuclease under the conditions where single-stranded DNA 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 <em>Hpa</em>I endonuclease digestion and the matching up of <em>Eco</em>RI-generated sticky end with <em>Escherichia coli</em> DNA polymerase I (Klenow fraction). However, the ligation efficiency of the S<sub>1</sub>-treated DNAs increased up to same level as the flush-ended DNA upon treatment with <em>E. coli</em> DNA polymerase I. Similar results were obtained in the case of S<sub>1</sub>-generated unit-length linear duplex DNA. S<sub>1</sub> does cleave both strands of superhelical DNA at unbasepaired sites.</p></div>\",\"PeriodicalId\":100164,\"journal\":{\"name\":\"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis\",\"volume\":\"656 1\",\"pages\":\"Pages 123-127\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1981-11-27\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/0005-2787(81)90035-6\",\"citationCount\":\"10\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/0005278781900356\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0005278781900356","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Efficiency of T4 DNA ligase-catalyzed end joining after S1 endonuclease treatment on duplex DNA containing single-stranded portions
Covalently closed-circular, superhelical SV40 DNA was used in all experiments. EcoRI endonuclease- and HpaII enonuclease-generated unit-length linear duplex DNAs were digested with S1 endonuclease under the conditions where single-stranded DNA 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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