Standardization of genetic transformation protocols in Escherichia coli and Agrobacterium tumefaciens for the generation of a collection of gene constructs
An important tool in plant genetic engineering is the vectors or genic constructs that contain reporter genes, utilized to facilitate the standardization of genetic transformation protocols. The objective of this study was evaluate genetic transformation protocols for generate in a...
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| Format: | Online |
| Language: | spa |
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Universidad Pedagógica y Tecnológica de Colombia
2018
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| Online Access: | https://revistas.uptc.edu.co/index.php/ciencia_en_desarrollo/article/view/3861 |
| _version_ | 1801706331269234688 |
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| author | Gómez Arias, Linda Yhiset Gómez Daza, Silvia Núñez Zarantes, Víctor |
| author_facet | Gómez Arias, Linda Yhiset Gómez Daza, Silvia Núñez Zarantes, Víctor |
| author_sort | Gómez Arias, Linda Yhiset |
| collection | OJS |
| description | An important tool in plant genetic engineering is the vectors or genic constructs that contain reporter genes, utilized to facilitate the standardization of genetic transformation protocols. The objective of this study was evaluate genetic transformation protocols for generate in a quick and confinable manner, a base collection
of genic constructs in Escherichia coli strain DHα and Agrobacterium tumefaciens strains LBA 4404, EHA 105 y C58, using 25 binary vectors (pSK1019, pMP2482 y 23 de la serie pCAMBIA) that contain the reporter genes gfp and gus, by using the heat shock and electroporation techniques. The transformants confirmation was made by PCR amplification and by restriction enzymes assay with Eco RI and Xho I. These approaches allowed verifying the presence Successful of the 25 vectors inside the used bacteria. The results indicate that it is necessary to standardize the transformation protocols in the bacterial strains to be used because they do not all transform with the same conditions and this work can be a reference for the standardization in other laboratories of genetic transformation when working in a massive way. |
| format | Online |
| id | oai:oai.revistas.uptc.edu.co:article-3861 |
| institution | Revista Ciencia en Desarrollo |
| language | spa |
| publishDate | 2018 |
| publisher | Universidad Pedagógica y Tecnológica de Colombia |
| record_format | ojs |
| spelling | oai:oai.revistas.uptc.edu.co:article-38612022-06-15T17:03:29Z Standardization of genetic transformation protocols in Escherichia coli and Agrobacterium tumefaciens for the generation of a collection of gene constructs Estandarización de protocolos de tranformación genética en Escherichia coli y para la generación de una colección de constructos génicos Gómez Arias, Linda Yhiset Gómez Daza, Silvia Núñez Zarantes, Víctor Gen reportero electroporación constructo génico transformación genética Reporter gen Electroporation gene construct genetic transformation An important tool in plant genetic engineering is the vectors or genic constructs that contain reporter genes, utilized to facilitate the standardization of genetic transformation protocols. The objective of this study was evaluate genetic transformation protocols for generate in a quick and confinable manner, a base collection of genic constructs in Escherichia coli strain DHα and Agrobacterium tumefaciens strains LBA 4404, EHA 105 y C58, using 25 binary vectors (pSK1019, pMP2482 y 23 de la serie pCAMBIA) that contain the reporter genes gfp and gus, by using the heat shock and electroporation techniques. The transformants confirmation was made by PCR amplification and by restriction enzymes assay with Eco RI and Xho I. These approaches allowed verifying the presence Successful of the 25 vectors inside the used bacteria. The results indicate that it is necessary to standardize the transformation protocols in the bacterial strains to be used because they do not all transform with the same conditions and this work can be a reference for the standardization in other laboratories of genetic transformation when working in a massive way. Una herramienta necesaria en la ingeniería genética de plantas son los vectores o constructos génicos que contienen genes reporteros, pues facilitan la estandarización de procedimientos de transformación genética. El objetivo de éste estudio fue evaluar protocolos de transformación genética para generar unprocedimiento que permitiera de manera rápida y con able obtener una colección base de constructosgénicos en Escherichia coli cepa DH5α y Agrobacterium tumefaciens cepas LBA 4404, EHA 105 y C58, empleando 25 vectores vectores binarios (pSK1019, pMP2482 y 23 de la serie pCAMBIA) que contienen los genes reporteros gfp y gus utilizando las técnicas de choque térmico y electroporación. La con rma- ción de los transformantes se realizó mediante PCR y cortes con enzimas de restricción (Eco RI y Xho I),lo que permitió veri car la presencia exitosa de éstos 25 vectores dentro de las bacterias empleadas. Losresultados indican que es necesario estandarizar los protocolos de transformación en las cepas bacterianas a utilizar porque no todas transforman con las mismas condiciones y éste trabajo puede ser un referente para la estandarización en otros laboratorios de transformación genética cuando se trabaja de manera masiva. Universidad Pedagógica y Tecnológica de Colombia 2018-07-04 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Transformación genética bacteriana application/pdf https://revistas.uptc.edu.co/index.php/ciencia_en_desarrollo/article/view/3861 10.19053/01217488.v9.n2.2018.3861 Ciencia En Desarrollo; Vol. 9 No. 2 (2018): Vol 9, Núm. 2 (2018): Vol 9, Núm 2(2018): Julio- Diciembre; 9-16 Ciencia en Desarrollo; Vol. 9 Núm. 2 (2018): Vol 9, Núm. 2 (2018): Vol 9, Núm 2(2018): Julio- Diciembre; 9-16 2462-7658 0121-7488 spa https://revistas.uptc.edu.co/index.php/ciencia_en_desarrollo/article/view/3861/7244 Derechos de autor 2018 CIENCIA EN DESARROLLO |
| spellingShingle | Gen reportero electroporación constructo génico transformación genética Reporter gen Electroporation gene construct genetic transformation Gómez Arias, Linda Yhiset Gómez Daza, Silvia Núñez Zarantes, Víctor Standardization of genetic transformation protocols in Escherichia coli and Agrobacterium tumefaciens for the generation of a collection of gene constructs |
| title | Standardization of genetic transformation protocols in Escherichia coli and Agrobacterium tumefaciens for the generation of a collection of gene constructs |
| title_alt | Estandarización de protocolos de tranformación genética en Escherichia coli y para la generación de una colección de constructos génicos |
| title_full | Standardization of genetic transformation protocols in Escherichia coli and Agrobacterium tumefaciens for the generation of a collection of gene constructs |
| title_fullStr | Standardization of genetic transformation protocols in Escherichia coli and Agrobacterium tumefaciens for the generation of a collection of gene constructs |
| title_full_unstemmed | Standardization of genetic transformation protocols in Escherichia coli and Agrobacterium tumefaciens for the generation of a collection of gene constructs |
| title_short | Standardization of genetic transformation protocols in Escherichia coli and Agrobacterium tumefaciens for the generation of a collection of gene constructs |
| title_sort | standardization of genetic transformation protocols in escherichia coli and agrobacterium tumefaciens for the generation of a collection of gene constructs |
| topic | Gen reportero electroporación constructo génico transformación genética Reporter gen Electroporation gene construct genetic transformation |
| topic_facet | Gen reportero electroporación constructo génico transformación genética Reporter gen Electroporation gene construct genetic transformation |
| url | https://revistas.uptc.edu.co/index.php/ciencia_en_desarrollo/article/view/3861 |
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