Dissection of a Replication Origin of Xenopus DNA
A previously cloned 503-base pair (bp) EcoRI segment of genomic DNA from Xenopus laevis selected for enhancement of replication of its vector plasmid was moved to the EcoRI site of pBR322. This plasmid designated pJCC31 and five other clones, which were made by cleaving the 503-bp segment in relatio...
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| Veröffentlicht in: | Proc. Natl. Acad. Sci. U.S.A.; (United States) 1982-09, Vol.79 (18), p.5572-5576 |
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| creator | Chambers, Jasemine Choy Watanabe, Shinichi Taylor, J. Herbert |
| description | A previously cloned 503-base pair (bp) EcoRI segment of genomic DNA from Xenopus laevis selected for enhancement of replication of its vector plasmid was moved to the EcoRI site of pBR322. This plasmid designated pJCC31 and five other clones, which were made by cleaving the 503-bp segment in relation to a dispersed repeated sequence and subcloning, were compared with pBR322 for replication by microinjection into Xenopus eggs. The replication measured by incorporation of a32P-labeled nucleotide as well as semiconservative segregation and dilution of N6-methyladenine at the EcoRI sites showed pJCC31 to be about 15 times as efficient as pBR322. The next most efficient subclone, pJCC31-2, contains an insert with a complete 320-bp dispersed repeated sequence bracketed by an 8-bp direct repeat. This observation, along with our previous report that repeated sequences of the Alu family in the human genome enhanced replication of the vector plasmid nearly as much as that of the presumptive Xenopus origin, leads to the hypothesis that members of a subset of the short dispersed repeated sequences in vertebrates function as origins for chromosomal replication. Preliminary studies also show that the presumptive Xenopus origin contains a RNA polymerase promoter that increases the transcription of the plasmid when it is microinjected into Xenopus oocytes. |
| doi_str_mv | 10.1073/pnas.79.18.5572 |
| format | Article |
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Herbert</creator><creatorcontrib>Chambers, Jasemine Choy ; Watanabe, Shinichi ; Taylor, J. Herbert ; Florida State Univ., Tallahassee</creatorcontrib><description>A previously cloned 503-base pair (bp) EcoRI segment of genomic DNA from Xenopus laevis selected for enhancement of replication of its vector plasmid was moved to the EcoRI site of pBR322. This plasmid designated pJCC31 and five other clones, which were made by cleaving the 503-bp segment in relation to a dispersed repeated sequence and subcloning, were compared with pBR322 for replication by microinjection into Xenopus eggs. The replication measured by incorporation of a32P-labeled nucleotide as well as semiconservative segregation and dilution of N6-methyladenine at the EcoRI sites showed pJCC31 to be about 15 times as efficient as pBR322. The next most efficient subclone, pJCC31-2, contains an insert with a complete 320-bp dispersed repeated sequence bracketed by an 8-bp direct repeat. This observation, along with our previous report that repeated sequences of the Alu family in the human genome enhanced replication of the vector plasmid nearly as much as that of the presumptive Xenopus origin, leads to the hypothesis that members of a subset of the short dispersed repeated sequences in vertebrates function as origins for chromosomal replication. Preliminary studies also show that the presumptive Xenopus origin contains a RNA polymerase promoter that increases the transcription of the plasmid when it is microinjected into Xenopus oocytes.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.79.18.5572</identifier><identifier>PMID: 6752953</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>550401 - Genetics- Tracer Techniques ; AMPHIBIANS ; ANIMALS ; AQUATIC ORGANISMS ; Base Composition ; Base Sequence ; BASIC BIOLOGICAL SCIENCES ; BETA DECAY RADIOISOTOPES ; BETA-MINUS DECAY RADIOISOTOPES ; BIOLOGY ; Cells ; CLONING ; Cloning, Molecular ; DAYS LIVING RADIOISOTOPES ; DNA ; DNA - genetics ; DNA REPLICATION ; DNA-Directed RNA Polymerases - metabolism ; EFFICIENCY ; Eggs ; Escherichia coli - genetics ; Female ; FROGS ; Gels ; GENETICS ; ISOTOPE APPLICATIONS ; ISOTOPES ; LABELLING ; LIGHT NUCLEI ; NUCLEI ; Nucleic Acid Conformation ; Nucleic Acid Hybridization ; NUCLEIC ACID REPLICATION ; NUCLEIC ACIDS ; ODD-ODD NUCLEI ; Oocytes ; Oocytes - metabolism ; ORGANIC COMPOUNDS ; PHOSPHORUS 32 ; PHOSPHORUS ISOTOPES ; Plasmids ; Radioactive decay ; RADIOISOTOPES ; Replication origin ; RNA ; TRACER TECHNIQUES ; Transcription, Genetic ; VERTEBRATES ; Xenopus ; Xenopus laevis ; Yeasts</subject><ispartof>Proc. 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Herbert</creatorcontrib><creatorcontrib>Florida State Univ., Tallahassee</creatorcontrib><title>Dissection of a Replication Origin of Xenopus DNA</title><title>Proc. Natl. Acad. Sci. U.S.A.; (United States)</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>A previously cloned 503-base pair (bp) EcoRI segment of genomic DNA from Xenopus laevis selected for enhancement of replication of its vector plasmid was moved to the EcoRI site of pBR322. This plasmid designated pJCC31 and five other clones, which were made by cleaving the 503-bp segment in relation to a dispersed repeated sequence and subcloning, were compared with pBR322 for replication by microinjection into Xenopus eggs. The replication measured by incorporation of a32P-labeled nucleotide as well as semiconservative segregation and dilution of N6-methyladenine at the EcoRI sites showed pJCC31 to be about 15 times as efficient as pBR322. The next most efficient subclone, pJCC31-2, contains an insert with a complete 320-bp dispersed repeated sequence bracketed by an 8-bp direct repeat. This observation, along with our previous report that repeated sequences of the Alu family in the human genome enhanced replication of the vector plasmid nearly as much as that of the presumptive Xenopus origin, leads to the hypothesis that members of a subset of the short dispersed repeated sequences in vertebrates function as origins for chromosomal replication. Preliminary studies also show that the presumptive Xenopus origin contains a RNA polymerase promoter that increases the transcription of the plasmid when it is microinjected into Xenopus oocytes.</description><subject>550401 - Genetics- Tracer Techniques</subject><subject>AMPHIBIANS</subject><subject>ANIMALS</subject><subject>AQUATIC ORGANISMS</subject><subject>Base Composition</subject><subject>Base Sequence</subject><subject>BASIC BIOLOGICAL SCIENCES</subject><subject>BETA DECAY RADIOISOTOPES</subject><subject>BETA-MINUS DECAY RADIOISOTOPES</subject><subject>BIOLOGY</subject><subject>Cells</subject><subject>CLONING</subject><subject>Cloning, Molecular</subject><subject>DAYS LIVING RADIOISOTOPES</subject><subject>DNA</subject><subject>DNA - genetics</subject><subject>DNA REPLICATION</subject><subject>DNA-Directed RNA Polymerases - metabolism</subject><subject>EFFICIENCY</subject><subject>Eggs</subject><subject>Escherichia coli - genetics</subject><subject>Female</subject><subject>FROGS</subject><subject>Gels</subject><subject>GENETICS</subject><subject>ISOTOPE APPLICATIONS</subject><subject>ISOTOPES</subject><subject>LABELLING</subject><subject>LIGHT NUCLEI</subject><subject>NUCLEI</subject><subject>Nucleic Acid Conformation</subject><subject>Nucleic Acid Hybridization</subject><subject>NUCLEIC ACID REPLICATION</subject><subject>NUCLEIC ACIDS</subject><subject>ODD-ODD NUCLEI</subject><subject>Oocytes</subject><subject>Oocytes - metabolism</subject><subject>ORGANIC COMPOUNDS</subject><subject>PHOSPHORUS 32</subject><subject>PHOSPHORUS ISOTOPES</subject><subject>Plasmids</subject><subject>Radioactive decay</subject><subject>RADIOISOTOPES</subject><subject>Replication origin</subject><subject>RNA</subject><subject>TRACER TECHNIQUES</subject><subject>Transcription, Genetic</subject><subject>VERTEBRATES</subject><subject>Xenopus</subject><subject>Xenopus laevis</subject><subject>Yeasts</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1982</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1r3DAQxUVoSbdpz4FCy9JDe_JGY0ke6ZBDSPoRCA2UFnITsnacKHgtx5JD-9_Xm90mzaU5Dcz7vfngMbYPfAEcxUHfubRAswC9UArLHTYDbqCopOHP2IzzEgstS_mCvUzpmnNulOa7bLdCVRolZgxOQkrkc4jdPDZzN_9OfRu8u2ucD-Ey3PUvqIv9mOYn345eseeNaxO93tY99vPzpx_HX4uz8y-nx0dnhVdgcmEqp5dCo6trr4lMJYVroCbQygEqKepS1ks0wCsgYQRIEE0DlfS1KSWB2GOHm7n9WK9o6anLg2ttP4SVG37b6IJ9rHThyl7GWytkZWQ1-d9v_DHlYJMPmfyVj103fWsRhNSlmaAP2yVDvBkpZbsKyVPbuo7imCxKQEBUT4KgFEepcAIPNqAfYkoDNfcXA7fryOw6MovGgrbryCbH238fvee3GU36u62-Nv5VHw34-F_ANmPbZvqVJ_LNhrxOOQ4Pl5WIXPwBltaxzA</recordid><startdate>19820901</startdate><enddate>19820901</enddate><creator>Chambers, Jasemine Choy</creator><creator>Watanabe, Shinichi</creator><creator>Taylor, J. Herbert</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>OTOTI</scope><scope>5PM</scope></search><sort><creationdate>19820901</creationdate><title>Dissection of a Replication Origin of Xenopus DNA</title><author>Chambers, Jasemine Choy ; Watanabe, Shinichi ; Taylor, J. Herbert</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c519t-96a8d387abbc8ee9643af1be185a17543b24bd791061e3931413ff164cb924e13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1982</creationdate><topic>550401 - Genetics- Tracer Techniques</topic><topic>AMPHIBIANS</topic><topic>ANIMALS</topic><topic>AQUATIC ORGANISMS</topic><topic>Base Composition</topic><topic>Base Sequence</topic><topic>BASIC BIOLOGICAL SCIENCES</topic><topic>BETA DECAY RADIOISOTOPES</topic><topic>BETA-MINUS DECAY RADIOISOTOPES</topic><topic>BIOLOGY</topic><topic>Cells</topic><topic>CLONING</topic><topic>Cloning, Molecular</topic><topic>DAYS LIVING RADIOISOTOPES</topic><topic>DNA</topic><topic>DNA - genetics</topic><topic>DNA REPLICATION</topic><topic>DNA-Directed RNA Polymerases - metabolism</topic><topic>EFFICIENCY</topic><topic>Eggs</topic><topic>Escherichia coli - genetics</topic><topic>Female</topic><topic>FROGS</topic><topic>Gels</topic><topic>GENETICS</topic><topic>ISOTOPE APPLICATIONS</topic><topic>ISOTOPES</topic><topic>LABELLING</topic><topic>LIGHT NUCLEI</topic><topic>NUCLEI</topic><topic>Nucleic Acid Conformation</topic><topic>Nucleic Acid Hybridization</topic><topic>NUCLEIC ACID REPLICATION</topic><topic>NUCLEIC ACIDS</topic><topic>ODD-ODD NUCLEI</topic><topic>Oocytes</topic><topic>Oocytes - metabolism</topic><topic>ORGANIC COMPOUNDS</topic><topic>PHOSPHORUS 32</topic><topic>PHOSPHORUS ISOTOPES</topic><topic>Plasmids</topic><topic>Radioactive decay</topic><topic>RADIOISOTOPES</topic><topic>Replication origin</topic><topic>RNA</topic><topic>TRACER TECHNIQUES</topic><topic>Transcription, Genetic</topic><topic>VERTEBRATES</topic><topic>Xenopus</topic><topic>Xenopus laevis</topic><topic>Yeasts</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chambers, Jasemine Choy</creatorcontrib><creatorcontrib>Watanabe, Shinichi</creatorcontrib><creatorcontrib>Taylor, J. Herbert</creatorcontrib><creatorcontrib>Florida State Univ., Tallahassee</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>OSTI.GOV</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proc. Natl. Acad. Sci. U.S.A.; (United States)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chambers, Jasemine Choy</au><au>Watanabe, Shinichi</au><au>Taylor, J. Herbert</au><aucorp>Florida State Univ., Tallahassee</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Dissection of a Replication Origin of Xenopus DNA</atitle><jtitle>Proc. Natl. Acad. Sci. U.S.A.; (United States)</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1982-09-01</date><risdate>1982</risdate><volume>79</volume><issue>18</issue><spage>5572</spage><epage>5576</epage><pages>5572-5576</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>A previously cloned 503-base pair (bp) EcoRI segment of genomic DNA from Xenopus laevis selected for enhancement of replication of its vector plasmid was moved to the EcoRI site of pBR322. This plasmid designated pJCC31 and five other clones, which were made by cleaving the 503-bp segment in relation to a dispersed repeated sequence and subcloning, were compared with pBR322 for replication by microinjection into Xenopus eggs. The replication measured by incorporation of a32P-labeled nucleotide as well as semiconservative segregation and dilution of N6-methyladenine at the EcoRI sites showed pJCC31 to be about 15 times as efficient as pBR322. The next most efficient subclone, pJCC31-2, contains an insert with a complete 320-bp dispersed repeated sequence bracketed by an 8-bp direct repeat. This observation, along with our previous report that repeated sequences of the Alu family in the human genome enhanced replication of the vector plasmid nearly as much as that of the presumptive Xenopus origin, leads to the hypothesis that members of a subset of the short dispersed repeated sequences in vertebrates function as origins for chromosomal replication. Preliminary studies also show that the presumptive Xenopus origin contains a RNA polymerase promoter that increases the transcription of the plasmid when it is microinjected into Xenopus oocytes.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>6752953</pmid><doi>10.1073/pnas.79.18.5572</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Proc. Natl. Acad. Sci. U.S.A.; (United States), 1982-09, Vol.79 (18), p.5572-5576 |
| issn | 0027-8424 1091-6490 |
| language | eng |
| recordid | cdi_pnas_primary_79_18_5572 |
| source | Jstor Complete Legacy; MEDLINE; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | 550401 - Genetics- Tracer Techniques AMPHIBIANS ANIMALS AQUATIC ORGANISMS Base Composition Base Sequence BASIC BIOLOGICAL SCIENCES BETA DECAY RADIOISOTOPES BETA-MINUS DECAY RADIOISOTOPES BIOLOGY Cells CLONING Cloning, Molecular DAYS LIVING RADIOISOTOPES DNA DNA - genetics DNA REPLICATION DNA-Directed RNA Polymerases - metabolism EFFICIENCY Eggs Escherichia coli - genetics Female FROGS Gels GENETICS ISOTOPE APPLICATIONS ISOTOPES LABELLING LIGHT NUCLEI NUCLEI Nucleic Acid Conformation Nucleic Acid Hybridization NUCLEIC ACID REPLICATION NUCLEIC ACIDS ODD-ODD NUCLEI Oocytes Oocytes - metabolism ORGANIC COMPOUNDS PHOSPHORUS 32 PHOSPHORUS ISOTOPES Plasmids Radioactive decay RADIOISOTOPES Replication origin RNA TRACER TECHNIQUES Transcription, Genetic VERTEBRATES Xenopus Xenopus laevis Yeasts |
| title | Dissection of a Replication Origin of Xenopus DNA |
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