Use of RecA protein to enrich for homologous genes in a genomic library
- Stanford Univ. School of Medicine, CA (USA)
- Univ. of California, Los Angeles (USA)
- Yale Univ. School of Medicine, New Haven, CT (USA)
RecA protein-coated probe has been utilized to enrich genomic digests for desired genes in order to facilitate cloning from genomic libraries. Using a previously cloned HLA-B27 gene as the recA-coated enrichment probe, the authors obtained a mean 108x increase in the ratio of specific to nonspecific plaques in lambda libraries screened for B27 variant alleles of estimated 99% homology to the probe. Class I genes of lesser homology were less enriched. Loss of genomic DNA during the enrichment procedure can, however, restrict application of this technique whenever starting genomic DNA is very limited. Nevertheless, the impressive reduction in cloning effort and material makes recA enrichment a useful new tool for cloning homologous genes from genomic DNA.
- OSTI ID:
- 6810753
- Journal Information:
- Nucleic Acids Research; (UK), Vol. 16:16; ISSN 0305-1048
- Country of Publication:
- United States
- Language:
- English
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