{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,3,30]],"date-time":"2025-03-30T00:07:36Z","timestamp":1743293256537},"reference-count":58,"publisher":"Microbiology Society","issue":"10","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2006,10,1]]},"abstract":"<jats:p><jats:italic>Listeria monocytogenes<\/jats:italic>(Lm) is a Gram-positive intracellular pathogen that can elicit strong cellular immunity. An attenuated strain (Lmdd) with deletions in two genes (<jats:italic>dal<\/jats:italic>and<jats:italic>dat<\/jats:italic>) required for<jats:sc>d<\/jats:sc>-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<jats:sc>d<\/jats:sc>-alanine without compromising the safety of the original strain, the defect of Lmdd was complemented with a heterologous<jats:italic>Bacillus subtilis dal<\/jats:italic>gene, and the effects of truncating the upstream region of the gene on its transcription efficiency and of modifying its protein product with an<jats:italic>ssrA<\/jats:italic>tag at the 3\u2032-terminus were examined. The strains with 551\u200abp and 80\u200abp upstream regions showed high levels of transcription and grew without<jats:sc>d<\/jats:sc>-alanine. The strains with the shortest upstream regions, 48\u200abp and 18\u200abp, showed greatly decreased levels of transcription and failed to grow in the absence of<jats:sc>d<\/jats:sc>-alanine. Addition of an<jats:italic>ssrA<\/jats:italic>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<jats:sc>d<\/jats:sc>-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<jats:italic>L. monocytogenes<\/jats:italic>. These bacteria can thus induce immune responses in mice comparable to the original Lmdd strain, but without the need for exogenous<jats:sc>d<\/jats:sc>-alanine, and may have use as a live vaccine vector against infectious diseases and cancers.<\/jats:p>","DOI":"10.1099\/mic.0.28994-0","type":"journal-article","created":{"date-parts":[[2006,9,27]],"date-time":"2006-09-27T23:08:18Z","timestamp":1159398498000},"page":"3091-3102","source":"Crossref","is-referenced-by-count":7,"title":["A truncated Bacillus subtilis dal gene with a 3\u2032 ssrA gene tag regulates the growth and virulence of racemase-deficient Listeria monocytogenes"],"prefix":"10.1099","volume":"152","author":[{"given":"Zhongxia","family":"Li","sequence":"first","affiliation":[{"name":"Department of Microbiology, School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, 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