Abstract:
: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 Fosmids 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.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Kim UJ,Shizuya H,de Jong PJ,Birren B,Simon MIdoi
10.1093/nar/20.5.1083keywords:
subject
Has Abstractpub_date
1992-03-11 00:00:00pages
1083-5issue
5eissn
0305-1048issn
1362-4962journal_volume
20pub_type
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journal_title:Nucleic acids research
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