Construction and utility of a human chromosome 22-specific Fosmid library.
We have previously demonstrated the capability of the Fosmid vector based on Escherichia coli F-factor replicon to stably propagate cosmid-sized human genomic DNA fragments. Using the Fosmid vector, we have constructed and arrayed a 10 x human chromosome 22-specific library, partly by picking human positive clones from a total Fosmid library constructed using DNA from human-hamster hybrid cell line containing human chromosome 22, and partly by using flow-sorted chromosomal DNA. The clones and physical contig maps of the clones in the library will serve as a valuable resource for detailed analysis of the chromosome by providing reliable materials for high resolution mapping and sequencing. In order to efficiently build physical maps for the chromosomal regions of interest spanning several hundred kilobases to a megabase, it is necessary to rapidly identify subsets of the Fosmid clones from the library that cover such regions. In this report, we describe a method of using random amplification products derived from YAC clones to rapidly identify a subset of Fosmid clones that cover a specific genomic subregion.
Kim
U J
UJ
Division of Biology, California Institute of Technology, Pasadena 91125, USA.
Shizuya
H
H
Sainz
J
J
Garnes
J
J
Pulst
S M
SM
de Jong
P
P
Simon
M I
MI
D000818:Animals; D001483:Base Sequence; D018244:Chromosomes, Artificial, Yeast; D002876:Chromosomes, Bacterial; D002892:Chromosomes, Human, Pair 22; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D005822:Genetic Vectors; D015894:Genome, Human; D006801:Humans; D006822:Hybrid Cells; D017404:In Situ Hybridization, Fluorescence; D008969:Molecular Sequence Data