Increasing plasmid transformation efficiency of natural spizizen method in Bacillus Subtilis by a cell permeable peptide
Introduction: Some of bacterial species are able to uptake DNA molecule from environment, the yield of this process depends on some conditions such as plasmid size and host type. In the case of Bacillus subtilis, DNA uptake has low efficacy. Using Spizizen minimal medium is common method in plasmid...
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| Format: | Article |
| Language: | English |
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University of Isfahan
2013-01-01
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| Series: | Biological Journal of Microorganism |
| Subjects: | |
| Online Access: | http://uijs.ui.ac.ir/bjm/browse.php?a_id=101&slc_lang=fa&sid=1&ftxt=1 |
| _version_ | 1818448571524972544 |
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| author | Mehrdad Moosazadeh Moghaddam Mohammad Eftekhari Mohammad Heiat Hossein Aghamollaei |
| author_facet | Mehrdad Moosazadeh Moghaddam Mohammad Eftekhari Mohammad Heiat Hossein Aghamollaei |
| author_sort | Mehrdad Moosazadeh Moghaddam |
| collection | DOAJ |
| description | Introduction: Some of bacterial species are able to uptake DNA molecule from environment, the yield of this process depends on some conditions such as plasmid size and host type. In the case of Bacillus subtilis, DNA uptake has low efficacy. Using Spizizen minimal medium is common method in plasmid transformation into B. subtilis, but rate of this process is not suitable and noteworthy. The aim of this study was investigation of novel method for improvement of DNA transformation into B. subtilis based on CM11 cationic peptide as a membrane permeable agent.Materials and methods: In this study, for optimization of pWB980 plasmid transformation into B. subtilis, the CM11 cationic peptide was used. For this purpose, B. subtilis competent cell preparation in the present of different concentration of peptide was implemented by two methods. In the first method, after treatment of bacteria with different amount of peptide for 14h, plasmid was added. In the second method, several concentration of peptide with plasmid was exposed to bacteria simultaneously. Bacteria that uptake DNA were screened on LB agar medium containing kanamycin. The total transformed bacteria per microgram of DNA was calculated and compared with the control.Results: Plasmid transformation in best conditions was 6.5 folds higher than the control. This result was statistically significant (P value <0.001).Discussion and conclusion: This study showed that CM11 cationic peptide as a membrane permeable agent was able to increase plasmid transformation rate into B. subtilis. This property was useful for resolution of low transformation efficacy. |
| first_indexed | 2024-12-14T20:21:38Z |
| format | Article |
| id | doaj.art-42d34f2fe2e5464e95371a6f3bb77a4a |
| institution | Directory Open Access Journal |
| issn | 2322-5173 2322-5181 |
| language | English |
| last_indexed | 2024-12-14T20:21:38Z |
| publishDate | 2013-01-01 |
| publisher | University of Isfahan |
| record_format | Article |
| series | Biological Journal of Microorganism |
| spelling | doaj.art-42d34f2fe2e5464e95371a6f3bb77a4a2022-12-21T22:48:44ZengUniversity of IsfahanBiological Journal of Microorganism2322-51732322-51812013-01-01263140Increasing plasmid transformation efficiency of natural spizizen method in Bacillus Subtilis by a cell permeable peptideMehrdad Moosazadeh MoghaddamMohammad EftekhariMohammad HeiatHossein AghamollaeiIntroduction: Some of bacterial species are able to uptake DNA molecule from environment, the yield of this process depends on some conditions such as plasmid size and host type. In the case of Bacillus subtilis, DNA uptake has low efficacy. Using Spizizen minimal medium is common method in plasmid transformation into B. subtilis, but rate of this process is not suitable and noteworthy. The aim of this study was investigation of novel method for improvement of DNA transformation into B. subtilis based on CM11 cationic peptide as a membrane permeable agent.Materials and methods: In this study, for optimization of pWB980 plasmid transformation into B. subtilis, the CM11 cationic peptide was used. For this purpose, B. subtilis competent cell preparation in the present of different concentration of peptide was implemented by two methods. In the first method, after treatment of bacteria with different amount of peptide for 14h, plasmid was added. In the second method, several concentration of peptide with plasmid was exposed to bacteria simultaneously. Bacteria that uptake DNA were screened on LB agar medium containing kanamycin. The total transformed bacteria per microgram of DNA was calculated and compared with the control.Results: Plasmid transformation in best conditions was 6.5 folds higher than the control. This result was statistically significant (P value <0.001).Discussion and conclusion: This study showed that CM11 cationic peptide as a membrane permeable agent was able to increase plasmid transformation rate into B. subtilis. This property was useful for resolution of low transformation efficacy.http://uijs.ui.ac.ir/bjm/browse.php?a_id=101&slc_lang=fa&sid=1&ftxt=1TransformationCM11 PeptidepWB980Bacillus subtilis |
| spellingShingle | Mehrdad Moosazadeh Moghaddam Mohammad Eftekhari Mohammad Heiat Hossein Aghamollaei Increasing plasmid transformation efficiency of natural spizizen method in Bacillus Subtilis by a cell permeable peptide Biological Journal of Microorganism Transformation CM11 Peptide pWB980 Bacillus subtilis |
| title | Increasing plasmid transformation efficiency of natural spizizen method in Bacillus Subtilis by a cell permeable peptide |
| title_full | Increasing plasmid transformation efficiency of natural spizizen method in Bacillus Subtilis by a cell permeable peptide |
| title_fullStr | Increasing plasmid transformation efficiency of natural spizizen method in Bacillus Subtilis by a cell permeable peptide |
| title_full_unstemmed | Increasing plasmid transformation efficiency of natural spizizen method in Bacillus Subtilis by a cell permeable peptide |
| title_short | Increasing plasmid transformation efficiency of natural spizizen method in Bacillus Subtilis by a cell permeable peptide |
| title_sort | increasing plasmid transformation efficiency of natural spizizen method in bacillus subtilis by a cell permeable peptide |
| topic | Transformation CM11 Peptide pWB980 Bacillus subtilis |
| url | http://uijs.ui.ac.ir/bjm/browse.php?a_id=101&slc_lang=fa&sid=1&ftxt=1 |
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