Stable propagation of cosmid sized human DNA inserts in an F factor based vector

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Abstract
Instability ofcomplex mammalian genomic DNAinserts iscommonplace incosmidlibraries constructed in conventional multicopy vectors. Todevelop ameans toconstruct stable libraries, wehavedeveloped alow copynumber cosmid vector based ontheE.coll Ffactor replicon (Fosmid). Wehavetested relative stability of humanDNAinserts inFosmids andintwoconventional multicopy vectors (Lawrist 16andSupercos) by comparing thefrequency ofchanges inrestriction patterns oftheinserts after propagating randomly picked humangenomic clones based onthese vectors. We foundthattheclones basedonFosmidvector undergo detectable changesata greatly reduced frequency. We alsoobserved thatsequences that undergo drastic rearrangements anddeletions during propagation ina conventional vector werestably propagated whenrecloned asFosmids. Theresults indicate thatFosmidsystemmay be useful for constructing stable libraries fromcomplex genomes.