Expression of the rice hoja blanca virus (RHBV) non-structural protein 3 (NS3) in Escherichia coli and its in situ localization in RHBV-infected rice tissues
Expression of the rice hoja blanca virus (RHBV) non-structural protein 3 (NS3) in Escherichia coli and its in situ localization in RHBV-infected rice tissues
| dc.creator | Muñoz, Miguel | |
| dc.creator | Bolaños, Isela | |
| dc.creator | Arrieta-Espinoza, Griselda | |
| dc.creator | Espinoza, Ana M | |
| dc.date | 2004-09-01 | |
| dc.date.accessioned | 2023-08-03T17:51:19Z | |
| dc.date.available | 2023-08-03T17:51:19Z | |
| dc.identifier | https://revistas.ucr.ac.cr/index.php/rbt/article/view/15413 | |
| dc.identifier.uri | https://repositorioslatinoamericanos.uchile.cl/handle/2250/7897281 | |
| dc.description | The non-structural NS3 protein gene from the rice hoja blanca virus (RHBV) was fused to the glutathione- S-transferase carboxilic end and expressed in Escherichia coli strain JM83. Large quantities of fusion protein were produced in insoluble form. The fusion protein was fractionated in SDS-PAGE and purified by electroelution, polyclonal antibodies were raised in rabbit and the antiserum was absorbed with bacterial crude extract. A band of similar size as that of NS3 protein was observed in Western blots using extracts from RHBVinfected rice plants. Immunoelectron microscopy with colloidal gold-labeled antibodies against NS3 protein and the viral nucleocapsid protein revealed in situ accumulation of NS3 protein in the cytoplasm but not in the viral inclusion bodies, vacuoles or chloroplasts of RHBV-infected plants, following the same pattern of distribution as the RHBV nucleocapsid protein. | en-US |
| dc.description | El gen que codifica por la proteína no estructural NS3 del virus de la hoja blanca de arroz (RHBV) se fusionó al extremo carboxilo del gen de la glutationa-S-transferasa y se expresó en la cepa JM83 de Escherichia coli. Se obtuvieron altas concentraciones de la proteína de fusion (GST-NS3) en forma insoluble. La proteína de fusión se fraccionó en geles de SDS-PAGE, se purificó por electroelución, y se utilizó para producir anticuerpos policlonales en conejo . El antisuero producido se absorbió con extractos crudos de E. coli. Extractos crudos de plantas de arroz sanas e infectadas con el RHBV se evaluaron por Western blots detectándose una banda de peso molecular similar al estimado para la proteína NS3 (23KDa) en las plantas infectadas con el virus. Los tejidos provenientes de plantas infectadas con el RHBV se analizaron por medio de microscopia inmunoelectrónica con oro colloidal marcado con anticuerpos contra la proteína NS3 y la nucleoproteína viral N. Se observó una acumulación in situ de la proteína NS3 en el citoplasma, pero no se detectó en los cuerpos de inclusion, vacuolas o cloroplastos. Se demostró que la proteína NS3 sigue el mismo patron de distribución que el de la nucleoproteína viral N del RHBV | es-ES |
| dc.format | application/pdf | |
| dc.language | eng | |
| dc.publisher | Universidad de Costa Rica | en-US |
| dc.relation | https://revistas.ucr.ac.cr/index.php/rbt/article/view/15413/14778 | |
| dc.rights | Derechos de autor 2004 Revista de Biología Tropical | es-ES |
| dc.rights | http://creativecommons.org/licenses/by/4.0 | es-ES |
| dc.source | Revista de Biología Tropical; Vol. 52 No. 3 (2004): Volume 52 - Regular number 3 - September 2004; 765–775 | en-US |
| dc.source | Revista de Biología Tropical; Vol. 52 Núm. 3 (2004): Volumen 52 - Número regular 3 - Setiembre 2004; 765–775 | es-ES |
| dc.source | Revista Biología Tropical; Vol. 52 N.º 3 (2004): Volumen 52 - Número regular 3 - Setiembre 2004; 765–775 | pt-PT |
| dc.source | 2215-2075 | |
| dc.source | 0034-7744 | |
| dc.source | 10.15517/rbt.v1i2 | |
| dc.subject | rice hoja blanca virus | en-US |
| dc.subject | RHBV | en-US |
| dc.subject | NS3 | en-US |
| dc.subject | polyclonal antibodies | en-US |
| dc.subject | immunoelectron microscopy | en-US |
| dc.subject | virus hoja blanca | es-ES |
| dc.subject | arroz | es-ES |
| dc.subject | RHBV | es-ES |
| dc.subject | NS3 | es-ES |
| dc.subject | anticuerpos policlonales | es-ES |
| dc.subject | inmunoelectro microscopia | es-ES |
| dc.title | Expression of the rice hoja blanca virus (RHBV) non-structural protein 3 (NS3) in Escherichia coli and its in situ localization in RHBV-infected rice tissues | en-US |
| dc.title | Expression of the rice hoja blanca virus (RHBV) non-structural protein 3 (NS3) in Escherichia coli and its in situ localization in RHBV-infected rice tissues | es-ES |
| dc.type | info:eu-repo/semantics/article | |
| dc.type | info:eu-repo/semantics/publishedVersion | |
| dc.type | Article | en-US |