Cre-mediated recombination at the murine whey acidic protein (MWAP) locus /
The mouse whey acidic protein (WAP) gene has been targeted in
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| Format: | Thesis Book |
| Language: | English |
| Published: |
[Place of publication not identified] :
[publisher not identified] ;
1996.
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| Subjects: | |
| Online Access: | http://proxy.library.tamu.edu/login?url=http://proquest.umi.com/pqdweb?did=739667691&sid=1&Fmt=2&clientId=2945&RQT=309&VName=PQD |
| Summary: | The mouse whey acidic protein (WAP) gene has been targeted in mouse embryonic stem (ES) cells. A replacement-type targeting vector, pWPNT, which contained a loxP-flanked neomycin phosphotransferase-thymidine kinase (neo-tk) cassette inserted into exon 4, had 3.Okb of 5' homology and 1.5kb of 3' homology with WAP. Targeted ES cell colonies were identified by Southern analysis; out of 199 colonies analyzed, 51 were positive (1:4). A cre-encoding plasmid, pBS 185, was electroporated into a targeted cell line to cause the deletion of the neo-tk cassette, leaving behind a single loxP tag. Modified ES cell colonies were identified by polymerase chain reaction (PCR); 44 out of 50 colonies (88%) had undergone cre-mediated deletion. A deletion cell line was electroporated with a cre-encoding plasmid and loxP- containing Neo plasmid for site-specific insertion into the chromosomally positioned loxP site. The frequency of the event was 23% (11 of 48) of that obtained with random integration as confirmed by Southern blot analysis. This demonstrates the feasibility of using the cre-loxP system for site-specific integration into modified loxP- tagged ES cells. |
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| Item Description: | Vita. "Major Subject: Veterinary Anatomy". Library's copy imperfect: page 139 containing Vita is wanting. |
| Physical Description: | xii, 138 leaves : illustrations ; 28 cm. Issued also on microfiche from University Microfilms Inc. |
| Bibliography: | Includes bibliographical references: pages 118-134. |