1887

Abstract

In this paper, the construction and evaluation of a chromosomal expression platform (CEP), which allows controlled gene expression following ectopic integration into the chromosome of , is described. CEP is based on the well-studied maltosaccharide-inducible system. To facilitate integration at CEP, a plasmid, pCEP, capable of replication in , but not in , was assembled. This plasmid contains an expression/selection cassette flanked on each side by more than 2 kb of pneumococcal DNA. The cassette comprises a maltose-inducible promoter, P, separated from a kanamycin-resistance gene by I and HI cloning sites. Clones harbouring the gene of interest integrated at CEP under the control of P can be obtained through direct transformation of an recipient with ligation products between that gene and I/HI-digested pCEP DNA, followed by selection for kanamycin-resistant transformants.

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2006-02-01
2020-04-03
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