Factors influencing electroporation-mediated gene transfer to Stylosanthes guianensis (Aubl.) Sw. protoplasts
In order to develop a high-efficiency and reproducible transformation protocol for Stylosanthes guianensis we assessed the biological and physical parameters affecting plant electroporation protoplasts. Energy input, as combinations of electric field strengths discharged by different capacitors, ele...
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| Format: | Article |
| Language: | English |
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Sociedade Brasileira de Genética
2002-01-01
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| Series: | Genetics and Molecular Biology |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572002000100014 |
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| author | Quecini V.M. Oliveira C.A. de Alves A.C. Vieira M.L.C. |
| author_facet | Quecini V.M. Oliveira C.A. de Alves A.C. Vieira M.L.C. |
| author_sort | Quecini V.M. |
| collection | DOAJ |
| description | In order to develop a high-efficiency and reproducible transformation protocol for Stylosanthes guianensis we assessed the biological and physical parameters affecting plant electroporation protoplasts. Energy input, as combinations of electric field strengths discharged by different capacitors, electroporation buffer and DNA form were evaluated. Transformation efficiency was assayed in vivo as transient reporter gene expression, using the GFP-coding gene mgfp5 driven by a CaMV 35S constitutive promoter. Energy input and electric field strength had a critical influence on transgene expression with higher transformation levels being achieved with 250 V.cm-1 discharged by 900 and 1000 muF capacitors. Linear plasmid DNA, the absence of chloride and the presence of calcium ions also increased transient gene expression, albeit not significantly. |
| first_indexed | 2024-12-10T14:59:20Z |
| format | Article |
| id | doaj.art-3dcc0982e17f4fdf9b7e287c9d4713ea |
| institution | Directory Open Access Journal |
| issn | 1415-4757 1678-4685 |
| language | English |
| last_indexed | 2024-12-10T14:59:20Z |
| publishDate | 2002-01-01 |
| publisher | Sociedade Brasileira de Genética |
| record_format | Article |
| series | Genetics and Molecular Biology |
| spelling | doaj.art-3dcc0982e17f4fdf9b7e287c9d4713ea2022-12-22T01:44:12ZengSociedade Brasileira de GenéticaGenetics and Molecular Biology1415-47571678-46852002-01-012517380Factors influencing electroporation-mediated gene transfer to Stylosanthes guianensis (Aubl.) Sw. protoplastsQuecini V.M.Oliveira C.A. deAlves A.C.Vieira M.L.C.In order to develop a high-efficiency and reproducible transformation protocol for Stylosanthes guianensis we assessed the biological and physical parameters affecting plant electroporation protoplasts. Energy input, as combinations of electric field strengths discharged by different capacitors, electroporation buffer and DNA form were evaluated. Transformation efficiency was assayed in vivo as transient reporter gene expression, using the GFP-coding gene mgfp5 driven by a CaMV 35S constitutive promoter. Energy input and electric field strength had a critical influence on transgene expression with higher transformation levels being achieved with 250 V.cm-1 discharged by 900 and 1000 muF capacitors. Linear plasmid DNA, the absence of chloride and the presence of calcium ions also increased transient gene expression, albeit not significantly.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572002000100014direct gene transferplant transformationforage legumeStylosanthes guianensis |
| spellingShingle | Quecini V.M. Oliveira C.A. de Alves A.C. Vieira M.L.C. Factors influencing electroporation-mediated gene transfer to Stylosanthes guianensis (Aubl.) Sw. protoplasts Genetics and Molecular Biology direct gene transfer plant transformation forage legume Stylosanthes guianensis |
| title | Factors influencing electroporation-mediated gene transfer to Stylosanthes guianensis (Aubl.) Sw. protoplasts |
| title_full | Factors influencing electroporation-mediated gene transfer to Stylosanthes guianensis (Aubl.) Sw. protoplasts |
| title_fullStr | Factors influencing electroporation-mediated gene transfer to Stylosanthes guianensis (Aubl.) Sw. protoplasts |
| title_full_unstemmed | Factors influencing electroporation-mediated gene transfer to Stylosanthes guianensis (Aubl.) Sw. protoplasts |
| title_short | Factors influencing electroporation-mediated gene transfer to Stylosanthes guianensis (Aubl.) Sw. protoplasts |
| title_sort | factors influencing electroporation mediated gene transfer to stylosanthes guianensis aubl sw protoplasts |
| topic | direct gene transfer plant transformation forage legume Stylosanthes guianensis |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572002000100014 |
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