1887

Abstract

SUMMARY: A series of plasmids has been constructed that can be used to fuse the β-galactosidase gene () of to chromosomal genes of . Insertion of the gene is facilitated by the use of a selectable chloramphenicol acetyl-transferase () gene. The latter is included, along with the gene, in a single DNA fragment or “cartridge” that can be removed from the plasmid with a variety of different restriction endonucleases. Methods applicable to any cloned gene are described that enable the cartridge to be inserted at specific sites, or at random, directly into the chromosome in a single step. These single-copy chromosomal fusions can be readily transferred, by selection for chloramphenicol resistance, to a temperate phage such as ϕ105, to permit a more extensive genetic analysis of the expression of the target gene. Alternatively, the cartridge and flanking DNA sequences can be transferred into different genetic backgrounds by transformation. These techniques have been used to construct, in a single step, fusions to genes in the sporulation operons and .

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1986-11-01
2021-07-23
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