Transgenic method based on RMCE technology
The invention discloses a transgenic method based on the RMCE technology. The transgenic method comprises the following steps: establishing a carrier by taking Rosa26 carrier as a framework and adding with Lox2272 locus or Attp70 locus; after correct sequencing of the carrier, electrotransferring to...
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| creator | WANG ZHENGLONG ZHENG DUNWU YU XIAOFENG LI FEIFENG SU CUI |
| description | The invention discloses a transgenic method based on the RMCE technology. The transgenic method comprises the following steps: establishing a carrier by taking Rosa26 carrier as a framework and adding with Lox2272 locus or Attp70 locus; after correct sequencing of the carrier, electrotransferring to an ES cell, and screening by adopting a culture medium containing antibiotics, thus obtaining a positive ES cell; verifying again by PCR or Southern Blot; transferring plasmids with Flp recombinase into the positive ES cell, thus obtaining a new positive ES cell; and administrating the positive ES cell after successful verification into the blastaea of a mouse through micro-injection, and then transferring into a surrogate mother mouse, thus obtaining an RMCE tool mouse with a fluorescent mark. The transgenic method is obtained by integrating RMCE and recombinase technology, thus having the advantages of the RMCE technology, and also being capable of realizing fixed-point implantation of genes, and moreover, the the test cycle is short, the efficiency is high, and the toxicity caused by random insertion of the genes is also reduced. |
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The transgenic method comprises the following steps: establishing a carrier by taking Rosa26 carrier as a framework and adding with Lox2272 locus or Attp70 locus; after correct sequencing of the carrier, electrotransferring to an ES cell, and screening by adopting a culture medium containing antibiotics, thus obtaining a positive ES cell; verifying again by PCR or Southern Blot; transferring plasmids with Flp recombinase into the positive ES cell, thus obtaining a new positive ES cell; and administrating the positive ES cell after successful verification into the blastaea of a mouse through micro-injection, and then transferring into a surrogate mother mouse, thus obtaining an RMCE tool mouse with a fluorescent mark. The transgenic method is obtained by integrating RMCE and recombinase technology, thus having the advantages of the RMCE technology, and also being capable of realizing fixed-point implantation of genes, and moreover, the the test cycle is short, the efficiency is high, and the toxicity caused by random insertion of the genes is also reduced.</description><language>eng</language><subject>AGRICULTURE ; ANIMAL HUSBANDRY ; BEER ; BIOCHEMISTRY ; CARE OF BIRDS, FISHES, INSECTS ; CHEMISTRY ; COMPOSITIONS THEREOF ; CULTURE MEDIA ; ENZYMOLOGY ; FISHING ; FORESTRY ; HUMAN NECESSITIES ; HUNTING ; METALLURGY ; MICROBIOLOGY ; MICROORGANISMS OR ENZYMES ; MUTATION OR GENETIC ENGINEERING ; NEW BREEDS OF ANIMALS ; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS ; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR ; SPIRITS ; TRAPPING ; VINEGAR ; WINE</subject><creationdate>2015</creationdate><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://worldwide.espacenet.com/publicationDetails/biblio?FT=D&date=20151118&DB=EPODOC&CC=CN&NR=105063087A$$EHTML$$P50$$Gepo$$Hfree_for_read</linktohtml><link.rule.ids>230,308,780,885,25562,76317</link.rule.ids><linktorsrc>$$Uhttps://worldwide.espacenet.com/publicationDetails/biblio?FT=D&date=20151118&DB=EPODOC&CC=CN&NR=105063087A$$EView_record_in_European_Patent_Office$$FView_record_in_$$GEuropean_Patent_Office$$Hfree_for_read</linktorsrc></links><search><creatorcontrib>WANG ZHENGLONG</creatorcontrib><creatorcontrib>ZHENG DUNWU</creatorcontrib><creatorcontrib>YU XIAOFENG</creatorcontrib><creatorcontrib>LI FEIFENG</creatorcontrib><creatorcontrib>SU CUI</creatorcontrib><title>Transgenic method based on RMCE technology</title><description>The invention discloses a transgenic method based on the RMCE technology. The transgenic method comprises the following steps: establishing a carrier by taking Rosa26 carrier as a framework and adding with Lox2272 locus or Attp70 locus; after correct sequencing of the carrier, electrotransferring to an ES cell, and screening by adopting a culture medium containing antibiotics, thus obtaining a positive ES cell; verifying again by PCR or Southern Blot; transferring plasmids with Flp recombinase into the positive ES cell, thus obtaining a new positive ES cell; and administrating the positive ES cell after successful verification into the blastaea of a mouse through micro-injection, and then transferring into a surrogate mother mouse, thus obtaining an RMCE tool mouse with a fluorescent mark. The transgenic method is obtained by integrating RMCE and recombinase technology, thus having the advantages of the RMCE technology, and also being capable of realizing fixed-point implantation of genes, and moreover, the the test cycle is short, the efficiency is high, and the toxicity caused by random insertion of the genes is also reduced.</description><subject>AGRICULTURE</subject><subject>ANIMAL HUSBANDRY</subject><subject>BEER</subject><subject>BIOCHEMISTRY</subject><subject>CARE OF BIRDS, FISHES, INSECTS</subject><subject>CHEMISTRY</subject><subject>COMPOSITIONS THEREOF</subject><subject>CULTURE MEDIA</subject><subject>ENZYMOLOGY</subject><subject>FISHING</subject><subject>FORESTRY</subject><subject>HUMAN NECESSITIES</subject><subject>HUNTING</subject><subject>METALLURGY</subject><subject>MICROBIOLOGY</subject><subject>MICROORGANISMS OR ENZYMES</subject><subject>MUTATION OR GENETIC ENGINEERING</subject><subject>NEW BREEDS OF ANIMALS</subject><subject>PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS</subject><subject>REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR</subject><subject>SPIRITS</subject><subject>TRAPPING</subject><subject>VINEGAR</subject><subject>WINE</subject><fulltext>true</fulltext><rsrctype>patent</rsrctype><creationdate>2015</creationdate><recordtype>patent</recordtype><sourceid>EVB</sourceid><recordid>eNrjZNAKKUrMK05PzctMVshNLcnIT1FISixOTVHIz1MI8nV2VShJTc7Iy8_JT6_kYWBNS8wpTuWF0twMim6uIc4euqkF-fGpxQWJyal5qSXxzn6GBqYGZsYGFuaOxsSoAQD5KSh7</recordid><startdate>20151118</startdate><enddate>20151118</enddate><creator>WANG ZHENGLONG</creator><creator>ZHENG DUNWU</creator><creator>YU XIAOFENG</creator><creator>LI FEIFENG</creator><creator>SU CUI</creator><scope>EVB</scope></search><sort><creationdate>20151118</creationdate><title>Transgenic method based on RMCE technology</title><author>WANG ZHENGLONG ; ZHENG DUNWU ; YU XIAOFENG ; LI FEIFENG ; SU CUI</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-epo_espacenet_CN105063087A3</frbrgroupid><rsrctype>patents</rsrctype><prefilter>patents</prefilter><language>eng</language><creationdate>2015</creationdate><topic>AGRICULTURE</topic><topic>ANIMAL HUSBANDRY</topic><topic>BEER</topic><topic>BIOCHEMISTRY</topic><topic>CARE OF BIRDS, FISHES, INSECTS</topic><topic>CHEMISTRY</topic><topic>COMPOSITIONS THEREOF</topic><topic>CULTURE MEDIA</topic><topic>ENZYMOLOGY</topic><topic>FISHING</topic><topic>FORESTRY</topic><topic>HUMAN NECESSITIES</topic><topic>HUNTING</topic><topic>METALLURGY</topic><topic>MICROBIOLOGY</topic><topic>MICROORGANISMS OR ENZYMES</topic><topic>MUTATION OR GENETIC ENGINEERING</topic><topic>NEW BREEDS OF ANIMALS</topic><topic>PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS</topic><topic>REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR</topic><topic>SPIRITS</topic><topic>TRAPPING</topic><topic>VINEGAR</topic><topic>WINE</topic><toplevel>online_resources</toplevel><creatorcontrib>WANG ZHENGLONG</creatorcontrib><creatorcontrib>ZHENG DUNWU</creatorcontrib><creatorcontrib>YU XIAOFENG</creatorcontrib><creatorcontrib>LI FEIFENG</creatorcontrib><creatorcontrib>SU CUI</creatorcontrib><collection>esp@cenet</collection></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext_linktorsrc</fulltext></delivery><addata><au>WANG ZHENGLONG</au><au>ZHENG DUNWU</au><au>YU XIAOFENG</au><au>LI FEIFENG</au><au>SU CUI</au><format>patent</format><genre>patent</genre><ristype>GEN</ristype><title>Transgenic method based on RMCE technology</title><date>2015-11-18</date><risdate>2015</risdate><abstract>The invention discloses a transgenic method based on the RMCE technology. The transgenic method comprises the following steps: establishing a carrier by taking Rosa26 carrier as a framework and adding with Lox2272 locus or Attp70 locus; after correct sequencing of the carrier, electrotransferring to an ES cell, and screening by adopting a culture medium containing antibiotics, thus obtaining a positive ES cell; verifying again by PCR or Southern Blot; transferring plasmids with Flp recombinase into the positive ES cell, thus obtaining a new positive ES cell; and administrating the positive ES cell after successful verification into the blastaea of a mouse through micro-injection, and then transferring into a surrogate mother mouse, thus obtaining an RMCE tool mouse with a fluorescent mark. The transgenic method is obtained by integrating RMCE and recombinase technology, thus having the advantages of the RMCE technology, and also being capable of realizing fixed-point implantation of genes, and moreover, the the test cycle is short, the efficiency is high, and the toxicity caused by random insertion of the genes is also reduced.</abstract><oa>free_for_read</oa></addata></record> |
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| subjects | AGRICULTURE ANIMAL HUSBANDRY BEER BIOCHEMISTRY CARE OF BIRDS, FISHES, INSECTS CHEMISTRY COMPOSITIONS THEREOF CULTURE MEDIA ENZYMOLOGY FISHING FORESTRY HUMAN NECESSITIES HUNTING METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING NEW BREEDS OF ANIMALS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR SPIRITS TRAPPING VINEGAR WINE |
| title | Transgenic method based on RMCE technology |
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