Kim, Ung-Jin and Shizuya, Hiroaki and de Jong, Pieter J. and Birren, Bruce and Simon, Melvin I. (1992) Stable propagation of cosmid sized human DNA inserts in an F factor based vector. Nucleic Acids Research, 20 (5). pp. 1083-1085. ISSN 0305-1048 http://resolver.caltech.edu/CaltechAUTHORS:KIMnar92
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Instability of complex mammalian genomic DNA inserts is commonplace in cosmid libraries constructed in conventional multicopy vectors. To develop a means to construct stable libraries, we have developed a low copy number cosmid vector based on the E.coli F factor replicon (Fosmid). We have tested relative stability of human DNA inserts in Fosmlds and in two conventional multicopy vectors (Lawrist 16 and Supercos) by comparing the frequency of changes In restriction patterns of the inserts after propagating randomly picked human genomic clones based on these vectors. We found that the clones based on Fosmid vector undergo detectable changes at a greatly reduced frequency. We also observed that sequences that undergo drastic rearrangements and deletions during propagation In a conventional vector were stably propagated when recloned as Fosmids. The results indicate that Fosmid system may be useful for constructing stable libraries from complex genomes.
|Additional Information:||© 1992. Received December 2, 1991; Accepted January 5, 1992. This work was supported by a grant from the Department of Energy, DE-FG03-89 ER60891 to M.I.S. Work by P.J. de Jong was supported by U.S. Department of Energy contract No.W-7405-ENG-48.|
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|Deposited On:||29 Oct 2007|
|Last Modified:||26 Dec 2012 09:45|
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