Plasmids for recombination-based screening
dc.contributor.author | Stewart, Gordon D. | en_US |
dc.contributor.author | Hauser, Michael A. | en_US |
dc.contributor.author | Kang, Harold | en_US |
dc.contributor.author | McCann, Damien P. | en_US |
dc.contributor.author | Osemlak, Matgorzata M. | en_US |
dc.contributor.author | Kurnit, David M. | en_US |
dc.contributor.author | Hanzlik, Andrzej J. | en_US |
dc.date.accessioned | 2006-04-10T14:34:50Z | |
dc.date.available | 2006-04-10T14:34:50Z | |
dc.date.issued | 1991-09-30 | en_US |
dc.identifier.citation | Stewart, Gordon D., Hauser, Michael A., Kang, Harold, McCann, Damien P., Osemlak, Matgorzata M., Kurnit, David M., Hanzlik, Andrzej J. (1991/09/30)."Plasmids for recombination-based screening." Gene 106(1): 97-101. <http://hdl.handle.net/2027.42/29122> | en_US |
dc.identifier.uri | http://www.sciencedirect.com/science/article/B6T39-47PNXWK-RG/2/c74cdede44b38ffb5bf755ec375d6385 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/29122 | |
dc.identifier.uri | http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=1834525&dopt=citation | en_US |
dc.description.abstract | To facilitate recombination-based screening, we constructed the ColE1-based plasmid, [pi]G4, that confers chloramphenicol resistance, contains a polylinker with multiple unique restriction enzyme recognition sequences, and contains the genetic marker, supF. To facilitate recombination-based screening followed by rapid DNA sequencing, we inserted the selectable marker, supF, into each of 20 high-copy-number (hcn) pUC-derived NoC plasmids that were designed for multiplex DNA sequencing. To facilitate recombination-based screening of common cDNA libraries that often contain ColE1 sequences, we constructed a supF-carrying plasmid whose replication was driven from an R6K replicon that does not share sequence homology with ColE1. Furthermore, we incorporated a useful polylinker and increased the copy number of this plasmid to create the 4.4-kb hcn plasmid, pMAD1. Thus, these plasmids allow: (1) background-free transformation of cells by a supF plasmid carrying an antibiotic-resistance marker; (2) simultaneous performance of the recombination-based assay and DNA sequencing; and (3) screening bacteriophage cDNA libraries that contain ColE1 sequences by recombination with a supF plasmid that is not homologous to ColE1 derivatives. | en_US |
dc.format.extent | 512085 bytes | |
dc.format.extent | 3118 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.language.iso | en_US | |
dc.publisher | Elsevier | en_US |
dc.title | Plasmids for recombination-based screening | en_US |
dc.type | Article | en_US |
dc.rights.robots | IndexNoFollow | en_US |
dc.subject.hlbsecondlevel | Molecular, Cellular and Developmental Biology | en_US |
dc.subject.hlbsecondlevel | Genetics | en_US |
dc.subject.hlbsecondlevel | Biological Chemistry | en_US |
dc.subject.hlbtoplevel | Science | en_US |
dc.subject.hlbtoplevel | Health Sciences | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Departments of Pediatrics and Human Genetics, Howard Hughes Medical Institute, University of Michigan Medical Center, Ann Arbor, MI 48109-0650, U.S.A. | en_US |
dc.contributor.affiliationum | Departments of Pediatrics and Human Genetics, Howard Hughes Medical Institute, University of Michigan Medical Center, Ann Arbor, MI 48109-0650, U.S.A. | en_US |
dc.contributor.affiliationum | Departments of Pediatrics and Human Genetics, Howard Hughes Medical Institute, University of Michigan Medical Center, Ann Arbor, MI 48109-0650, U.S.A. | en_US |
dc.contributor.affiliationum | Departments of Pediatrics and Human Genetics, Howard Hughes Medical Institute, University of Michigan Medical Center, Ann Arbor, MI 48109-0650, U.S.A. | en_US |
dc.contributor.affiliationum | Departments of Pediatrics and Human Genetics, Howard Hughes Medical Institute, University of Michigan Medical Center, Ann Arbor, MI 48109-0650, U.S.A. | en_US |
dc.contributor.affiliationum | Departments of Pediatrics and Human Genetics, Howard Hughes Medical Institute, University of Michigan Medical Center, Ann Arbor, MI 48109-0650, U.S.A. | en_US |
dc.contributor.affiliationum | Departments of Pediatrics and Human Genetics, Howard Hughes Medical Institute, University of Michigan Medical Center, Ann Arbor, MI 48109-0650, U.S.A. | en_US |
dc.identifier.pmid | 1834525 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/29122/1/0000161.pdf | en_US |
dc.identifier.doi | http://dx.doi.org/10.1016/0378-1119(91)90571-R | en_US |
dc.identifier.source | Gene | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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