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Nucleic Acids Research, 1990, Vol. 18, No. 4 725-731
© 1990

GENOME STRUCTURE AND MAPPING

Derivation of clones from the choroideremia locus by preparative field inversion gel electrophoresis

T.J.R. van de Pol, F.P.M. Cremers*, R.M. Brohet, B. Wieringa and H.-H. Ropers

Department of Human Genetics, Radboud Hospital, University of Nijmegen PO Box 9101, 6500HB Nijmegen, The Netherlands

*To whom correspondence should be addressed

Received December 14, 1989. Revised January 17, 1990. Accepted January 17, 1990.

By making use of preparative field inversion gel electrophoresis, we have constructed a lambdaZAP library that is highly enriched for sequences from the chorolderemia locus. In vivo excision of pBluescript SK(–) constructs from lambdaZAP obviates the subcloning of DNA inserts and allows for rapid processIng of several hundred recombinants. From 625 kb Sf/1 fragment we isolated 7 clones that were physically mapped using microdeletlons associated with the disease. One of these clones is located within, or Just telomeric to, the chorolderemia gene and detects two restriction fragment length polymorphisms (RFLPs). Another clone detects a RFLP which maps centromeric to the disease locus. Together these probes should Improve the reliability of linkage analysis in choroideremia families and should pave the way for the Isolation of the choroideremia gene.


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