<i>Lysinibacillus sphaericus</i> III(3)7 and Plasmid Vector pMK4: New Challenges in Cloning Platforms
The acquisition and especially the maintenance of a plasmid usually brings a fitness cost that reduces the reproductive rate of the bacterial host; for strains like <i>Lysinibacillus sphaericus</i> III(3)7, which possesses important environmental properties, this alteration along with mo...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-05-01
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| Series: | Microbiology Research |
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| Online Access: | https://www.mdpi.com/2036-7481/12/2/31 |
| _version_ | 1797535291758608384 |
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| author | Diana C. León Jenny Dussán |
| author_facet | Diana C. León Jenny Dussán |
| author_sort | Diana C. León |
| collection | DOAJ |
| description | The acquisition and especially the maintenance of a plasmid usually brings a fitness cost that reduces the reproductive rate of the bacterial host; for strains like <i>Lysinibacillus sphaericus</i> III(3)7, which possesses important environmental properties, this alteration along with morphological changes and reduced sporulation rates may exert a negative effect on metabolic studies using plasmids as cloning platforms. The aim of this study is to approach the metabolic behavior of pMK4-bearing cells of <i>L. sphaericus</i> III(3)7 through the use of bioinformatic and in vitro analyses. An incompatibility model between the pMK4 vector and a predicted megaplasmid, pBsph, inside III(3)7 cells was constructed based on an <i>incA</i> region. Additionally, in vitro long-term plasmid stability was not found in plasmid-bearing cells. Alignments between replicons, mobile genetic elements and RNA-RNA interactions were assessed, pairwise alignment visualization, graphic models and morphological changes were evaluated by SEM. Metabolite analysis was done through HPLC coupled to a Q-TOF 6545, and electrospray ionization was used, finally, <i>Aedes aegypti</i> and <i>Culex quinquefasciatus</i> larvae were used for larvicidal activity assessment. Results found, a decreased growth rate, spore formation reduction and morphological changes, which supported the idea of metabolic cost exerted by pMK4. An incompatibility between pMK4 and pBsph appears to take place inside <i>L. sphaericus</i> III(3)7 cells, however, further in vitro studies are needed to confirm it. |
| first_indexed | 2024-03-10T11:43:26Z |
| format | Article |
| id | doaj.art-77c2938910db4ae7aa83141eec1ab1b0 |
| institution | Directory Open Access Journal |
| issn | 2036-7481 |
| language | English |
| last_indexed | 2024-03-10T11:43:26Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microbiology Research |
| spelling | doaj.art-77c2938910db4ae7aa83141eec1ab1b02023-11-21T18:18:52ZengMDPI AGMicrobiology Research2036-74812021-05-0112245547910.3390/microbiolres12020031<i>Lysinibacillus sphaericus</i> III(3)7 and Plasmid Vector pMK4: New Challenges in Cloning PlatformsDiana C. León0Jenny Dussán1Centro de Investigaciones Microbiológicas (CIMIC), Department of Biological Sciences, Universidad de Los Andes, Bogotá 111711, ColombiaCentro de Investigaciones Microbiológicas (CIMIC), Department of Biological Sciences, Universidad de Los Andes, Bogotá 111711, ColombiaThe acquisition and especially the maintenance of a plasmid usually brings a fitness cost that reduces the reproductive rate of the bacterial host; for strains like <i>Lysinibacillus sphaericus</i> III(3)7, which possesses important environmental properties, this alteration along with morphological changes and reduced sporulation rates may exert a negative effect on metabolic studies using plasmids as cloning platforms. The aim of this study is to approach the metabolic behavior of pMK4-bearing cells of <i>L. sphaericus</i> III(3)7 through the use of bioinformatic and in vitro analyses. An incompatibility model between the pMK4 vector and a predicted megaplasmid, pBsph, inside III(3)7 cells was constructed based on an <i>incA</i> region. Additionally, in vitro long-term plasmid stability was not found in plasmid-bearing cells. Alignments between replicons, mobile genetic elements and RNA-RNA interactions were assessed, pairwise alignment visualization, graphic models and morphological changes were evaluated by SEM. Metabolite analysis was done through HPLC coupled to a Q-TOF 6545, and electrospray ionization was used, finally, <i>Aedes aegypti</i> and <i>Culex quinquefasciatus</i> larvae were used for larvicidal activity assessment. Results found, a decreased growth rate, spore formation reduction and morphological changes, which supported the idea of metabolic cost exerted by pMK4. An incompatibility between pMK4 and pBsph appears to take place inside <i>L. sphaericus</i> III(3)7 cells, however, further in vitro studies are needed to confirm it.https://www.mdpi.com/2036-7481/12/2/31incompatibilityplasmidmetabolic burdenantisense RNAsporulationlarvicidal activity |
| spellingShingle | Diana C. León Jenny Dussán <i>Lysinibacillus sphaericus</i> III(3)7 and Plasmid Vector pMK4: New Challenges in Cloning Platforms Microbiology Research incompatibility plasmid metabolic burden antisense RNA sporulation larvicidal activity |
| title | <i>Lysinibacillus sphaericus</i> III(3)7 and Plasmid Vector pMK4: New Challenges in Cloning Platforms |
| title_full | <i>Lysinibacillus sphaericus</i> III(3)7 and Plasmid Vector pMK4: New Challenges in Cloning Platforms |
| title_fullStr | <i>Lysinibacillus sphaericus</i> III(3)7 and Plasmid Vector pMK4: New Challenges in Cloning Platforms |
| title_full_unstemmed | <i>Lysinibacillus sphaericus</i> III(3)7 and Plasmid Vector pMK4: New Challenges in Cloning Platforms |
| title_short | <i>Lysinibacillus sphaericus</i> III(3)7 and Plasmid Vector pMK4: New Challenges in Cloning Platforms |
| title_sort | i lysinibacillus sphaericus i iii 3 7 and plasmid vector pmk4 new challenges in cloning platforms |
| topic | incompatibility plasmid metabolic burden antisense RNA sporulation larvicidal activity |
| url | https://www.mdpi.com/2036-7481/12/2/31 |
| work_keys_str_mv | AT dianacleon ilysinibacillussphaericusiiii37andplasmidvectorpmk4newchallengesincloningplatforms AT jennydussan ilysinibacillussphaericusiiii37andplasmidvectorpmk4newchallengesincloningplatforms |