Physical mapping of 32 genetic markers on the PAO1 chromosome Free

Abstract

The chromosome was fractionated with the enzymes and and genomic fragments were separated by PFGE and used for mapping a collection of 40 genes. This permitted the localization of 8 genes previously mapped and of 32 genes which had not been mapped. We showed that a careful search of databases and identification of sequences that were homologous to known genes could be used to design and synthesize DNA probes for the mapping of homologues by Southern hybridization with genomic fragments, resulting in definition of the locations of the and genetic markers. In addition, a combination of distinct DNA sources were utilized as radioactively labelled probes, including specific restriction fragments of the cloned genes ( DNA fragments prepared by PCR, and single-stranded DNA prepared from phagemid libraries that had been randomly sequenced. We used a PCR approach to clone fragments of the putative and genes. Random sequencing of DNA from phagemid libraries and database searching permitted the cloning of sequences from the and gene homologues. The described genomic methods permit the rapid mapping of the genome without linkage analysis.

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1996-01-01
2024-03-29
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