Simple and Rapid Method for PCR Characterization of Large Bacillus thuringiensis Strain Collections
Polymerase chain reaction (PCR)-based identification of Bacillus thuringiensis toxin genes has become a routine step in most B. thuringiensis isolation and characterization initiatives. In the present study we propose a simplified method for extensive PCR analysis of B. thuringiensis cry and cyt gen...
| Autores principales: | , , |
|---|---|
| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
2017
|
| Acceso en línea: | http://hdl.handle.net/20.500.11939/5053 |
| _version_ | 1855491859030212608 |
|---|---|
| author | Cristian Vidal-Quist, J. Castanera, Pedro González-Cabrera, Joel |
| author_browse | Castanera, Pedro Cristian Vidal-Quist, J. González-Cabrera, Joel |
| author_facet | Cristian Vidal-Quist, J. Castanera, Pedro González-Cabrera, Joel |
| author_sort | Cristian Vidal-Quist, J. |
| collection | ReDivia |
| description | Polymerase chain reaction (PCR)-based identification of Bacillus thuringiensis toxin genes has become a routine step in most B. thuringiensis isolation and characterization initiatives. In the present study we propose a simplified method for extensive PCR analysis of B. thuringiensis cry and cyt genes of particular interest in large-scale screening programs. Fifty B. thuringiensis strains were screened for the presence of genes of the cry1 subfamily. Identical results were obtained when our method was used in comparison to other methodology based on a standard alkaline lysis preparation of plasmid DNA. Additional tests evidenced the fitness of our method in a particular multiplex-PCR analysis. The main advantages of the proposed methodology are that it is simpler and quicker than commonly used protocols (as DNA template preparation is substituted by direct addition of small amounts of B. thuringiensis liquid cell suspension to the reaction mixture) and that it allows simultaneous handling (bacterial growth and PCR) of up to 96 strains per round. |
| format | Artículo |
| id | ReDivia5053 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia50532025-04-25T14:45:15Z Simple and Rapid Method for PCR Characterization of Large Bacillus thuringiensis Strain Collections Cristian Vidal-Quist, J. Castanera, Pedro González-Cabrera, Joel Polymerase chain reaction (PCR)-based identification of Bacillus thuringiensis toxin genes has become a routine step in most B. thuringiensis isolation and characterization initiatives. In the present study we propose a simplified method for extensive PCR analysis of B. thuringiensis cry and cyt genes of particular interest in large-scale screening programs. Fifty B. thuringiensis strains were screened for the presence of genes of the cry1 subfamily. Identical results were obtained when our method was used in comparison to other methodology based on a standard alkaline lysis preparation of plasmid DNA. Additional tests evidenced the fitness of our method in a particular multiplex-PCR analysis. The main advantages of the proposed methodology are that it is simpler and quicker than commonly used protocols (as DNA template preparation is substituted by direct addition of small amounts of B. thuringiensis liquid cell suspension to the reaction mixture) and that it allows simultaneous handling (bacterial growth and PCR) of up to 96 strains per round. 2017-06-01T10:11:36Z 2017-06-01T10:11:36Z 2009 MAY 2009 article Cristian Vidal-Quist, J., Castanera, P., Gonzalez-Cabrera, J. (2009). Simple and Rapid Method for PCR Characterization of Large Bacillus thuringiensis Strain Collections. Current microbiology, 58(5), 421-425. 0343-8651 http://hdl.handle.net/20.500.11939/5053 10.1007/s00284-008-9328-0 en openAccess Impreso |
| spellingShingle | Cristian Vidal-Quist, J. Castanera, Pedro González-Cabrera, Joel Simple and Rapid Method for PCR Characterization of Large Bacillus thuringiensis Strain Collections |
| title | Simple and Rapid Method for PCR Characterization of Large Bacillus thuringiensis Strain Collections |
| title_full | Simple and Rapid Method for PCR Characterization of Large Bacillus thuringiensis Strain Collections |
| title_fullStr | Simple and Rapid Method for PCR Characterization of Large Bacillus thuringiensis Strain Collections |
| title_full_unstemmed | Simple and Rapid Method for PCR Characterization of Large Bacillus thuringiensis Strain Collections |
| title_short | Simple and Rapid Method for PCR Characterization of Large Bacillus thuringiensis Strain Collections |
| title_sort | simple and rapid method for pcr characterization of large bacillus thuringiensis strain collections |
| url | http://hdl.handle.net/20.500.11939/5053 |
| work_keys_str_mv | AT cristianvidalquistj simpleandrapidmethodforpcrcharacterizationoflargebacillusthuringiensisstraincollections AT castanerapedro simpleandrapidmethodforpcrcharacterizationoflargebacillusthuringiensisstraincollections AT gonzalezcabrerajoel simpleandrapidmethodforpcrcharacterizationoflargebacillusthuringiensisstraincollections |