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Volume 152, Issue 10

A truncated gene with a 3′ gene tag regulates the growth and virulence of racemase-deficient Free

Abstract

(Lm) is a Gram-positive intracellular pathogen that can elicit strong cellular immunity. An attenuated strain (Lmdd) with deletions in two genes ( and ) required for -alanine synthesis and viability has been shown to induce long-lived protective systemic and mucosal immune responses in mice when administered in the presence of the required amino acid. To bypass the necessity for exogenous -alanine without compromising the safety of the original strain, the defect of Lmdd was complemented with a heterologous gene, and the effects of truncating the upstream region of the gene on its transcription efficiency and of modifying its protein product with an tag at the 3′-terminus were examined. The strains with 551 bp and 80 bp upstream regions showed high levels of transcription and grew without -alanine. The strains with the shortest upstream regions, 48 bp and 18 bp, showed greatly decreased levels of transcription and failed to grow in the absence of -alanine. Addition of an tag to the longer genes resulted in a somewhat altered growth pattern in media and a reduced plaque size on L2 fibroblasts. These bacteria contained low levels of racemase protein and reduced free pools of -alanine. One of the strains tested further, Lmdd/pA80S, was rapidly cleared from the spleens of infected mice but nevertheless induced a strong immune response that protected mice against challenge by wild-type . These bacteria can thus induce immune responses in mice comparable to the original Lmdd strain, but without the need for exogenous -alanine, and may have use as a live vaccine vector against infectious diseases and cancers.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): Cam, chloramphenicol , i.v., intravenously , LLO, listeriolysin O and Sm, streptomycin
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2006-10-01
2024-04-18
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A truncated Bacillus subtilis dal gene with a 3′ ssrA gene tag regulates the growth and virulence of racemase-deficient Listeria monocytogenes
Microbiology 152, 3091 (2006); https://doi.org/10.1099/mic.0.28994-0
/content/journal/micro/10.1099/mic.0.28994-0
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