Improving <it>Salmonella </it>vector with <it>rec </it>mutation to stabilize the DNA cargoes
<p>Abstract</p> <p>Background</p> <p><it>Salmonella </it>has been employed to deliver therapeutic molecules against cancer and infectious diseases. As the carrier for target gene(s), the cargo plasmid should be stable in the bacterial vector. Plasmid recombi...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2011-02-01
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| Series: | BMC Microbiology |
| Online Access: | http://www.biomedcentral.com/1471-2180/11/31 |
| _version_ | 1818757517302300672 |
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| author | Zhang Xin Kong Wei Brenneman Karen Wanda Soo-Young Zhang Xiangmin Roland Kenneth Curtiss Roy |
| author_facet | Zhang Xin Kong Wei Brenneman Karen Wanda Soo-Young Zhang Xiangmin Roland Kenneth Curtiss Roy |
| author_sort | Zhang Xin |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p><it>Salmonella </it>has been employed to deliver therapeutic molecules against cancer and infectious diseases. As the carrier for target gene(s), the cargo plasmid should be stable in the bacterial vector. Plasmid recombination has been reduced in <it>E. coli </it>by mutating several genes including the <it>recA</it>, <it>recE</it>, <it>recF </it>and <it>recJ</it>. However, to our knowledge, there have been no published studies of the effect of these or any other genes that play a role in plasmid recombination in <it>Salmonella enterica</it>.</p> <p>Results</p> <p>The effect of <it>recA</it>, <it>recF </it>and <it>recJ </it>deletions on DNA recombination was examined in three serotypes of <it>Salmonella enterica</it>. We found that (1) intraplasmid recombination between direct duplications was RecF-independent in Typhimurium and Paratyphi A, but could be significantly reduced in Typhi by a Δ<it>recA </it>or Δ<it>recF </it>mutation; (2) in all three <it>Salmonella </it>serotypes, both Δ<it>recA </it>and Δ<it>recF </it>mutations reduced intraplasmid recombination when a 1041 bp intervening sequence was present between the duplications; (3) Δ<it>recA </it>and Δ<it>recF </it>mutations resulted in lower frequencies of interplasmid recombination in Typhimurium and Paratyphi A, but not in Typhi; (4) in some cases, a Δ<it>recJ </it>mutation could reduce plasmid recombination but was less effective than Δ<it>recA </it>and Δ<it>recF </it>mutations. We also examined chromosome-related recombination. The frequencies of intrachromosomal recombination and plasmid integration into the chromosome were 2 and 3 logs lower than plasmid recombination frequencies in Rec<sup>+ </sup>strains. A Δ<it>recA </it>mutation reduced both intrachromosomal recombination and plasmid integration frequencies.</p> <p>Conclusions</p> <p>The Δ<it>recA </it>and Δ<it>recF </it>mutations can reduce plasmid recombination frequencies in <it>Salmonella enterica</it>, but the effect can vary between serovars. This information will be useful for developing <it>Salmonella </it>delivery vectors able to stably maintain plasmid cargoes for vaccine development and gene therapy.</p> |
| first_indexed | 2024-12-18T06:12:12Z |
| format | Article |
| id | doaj.art-d524ac4c3c284eafb91d895cdf26e221 |
| institution | Directory Open Access Journal |
| issn | 1471-2180 |
| language | English |
| last_indexed | 2024-12-18T06:12:12Z |
| publishDate | 2011-02-01 |
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| spelling | doaj.art-d524ac4c3c284eafb91d895cdf26e2212022-12-21T21:18:22ZengBMCBMC Microbiology1471-21802011-02-011113110.1186/1471-2180-11-31Improving <it>Salmonella </it>vector with <it>rec </it>mutation to stabilize the DNA cargoesZhang XinKong WeiBrenneman KarenWanda Soo-YoungZhang XiangminRoland KennethCurtiss Roy<p>Abstract</p> <p>Background</p> <p><it>Salmonella </it>has been employed to deliver therapeutic molecules against cancer and infectious diseases. As the carrier for target gene(s), the cargo plasmid should be stable in the bacterial vector. Plasmid recombination has been reduced in <it>E. coli </it>by mutating several genes including the <it>recA</it>, <it>recE</it>, <it>recF </it>and <it>recJ</it>. However, to our knowledge, there have been no published studies of the effect of these or any other genes that play a role in plasmid recombination in <it>Salmonella enterica</it>.</p> <p>Results</p> <p>The effect of <it>recA</it>, <it>recF </it>and <it>recJ </it>deletions on DNA recombination was examined in three serotypes of <it>Salmonella enterica</it>. We found that (1) intraplasmid recombination between direct duplications was RecF-independent in Typhimurium and Paratyphi A, but could be significantly reduced in Typhi by a Δ<it>recA </it>or Δ<it>recF </it>mutation; (2) in all three <it>Salmonella </it>serotypes, both Δ<it>recA </it>and Δ<it>recF </it>mutations reduced intraplasmid recombination when a 1041 bp intervening sequence was present between the duplications; (3) Δ<it>recA </it>and Δ<it>recF </it>mutations resulted in lower frequencies of interplasmid recombination in Typhimurium and Paratyphi A, but not in Typhi; (4) in some cases, a Δ<it>recJ </it>mutation could reduce plasmid recombination but was less effective than Δ<it>recA </it>and Δ<it>recF </it>mutations. We also examined chromosome-related recombination. The frequencies of intrachromosomal recombination and plasmid integration into the chromosome were 2 and 3 logs lower than plasmid recombination frequencies in Rec<sup>+ </sup>strains. A Δ<it>recA </it>mutation reduced both intrachromosomal recombination and plasmid integration frequencies.</p> <p>Conclusions</p> <p>The Δ<it>recA </it>and Δ<it>recF </it>mutations can reduce plasmid recombination frequencies in <it>Salmonella enterica</it>, but the effect can vary between serovars. This information will be useful for developing <it>Salmonella </it>delivery vectors able to stably maintain plasmid cargoes for vaccine development and gene therapy.</p>http://www.biomedcentral.com/1471-2180/11/31 |
| spellingShingle | Zhang Xin Kong Wei Brenneman Karen Wanda Soo-Young Zhang Xiangmin Roland Kenneth Curtiss Roy Improving <it>Salmonella </it>vector with <it>rec </it>mutation to stabilize the DNA cargoes BMC Microbiology |
| title | Improving <it>Salmonella </it>vector with <it>rec </it>mutation to stabilize the DNA cargoes |
| title_full | Improving <it>Salmonella </it>vector with <it>rec </it>mutation to stabilize the DNA cargoes |
| title_fullStr | Improving <it>Salmonella </it>vector with <it>rec </it>mutation to stabilize the DNA cargoes |
| title_full_unstemmed | Improving <it>Salmonella </it>vector with <it>rec </it>mutation to stabilize the DNA cargoes |
| title_short | Improving <it>Salmonella </it>vector with <it>rec </it>mutation to stabilize the DNA cargoes |
| title_sort | improving it salmonella it vector with it rec it mutation to stabilize the dna cargoes |
| url | http://www.biomedcentral.com/1471-2180/11/31 |
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