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Development of a highly efficient system for assessing recombinant gene expression in plant cell suspensions via Agrobacterium tumefaciens transformation
Author(s) -
Fuentes Alejandro,
Ramos Pedro Luis,
Ayra Camilo,
Rodríguez Meilyn,
Ramírez Nadia,
Pujol Merardo
Publication year - 2004
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20030192
Subject(s) - cauliflower mosaic virus , bacillus thuringiensis , biology , agrobacterium , gene , gene expression , tobacco mosaic virus , western blot , microbiology and biotechnology , monoclonal antibody , virology , virus , antibody , genetically modified crops , transgene , bacteria , genetics
A transient gene‐expression system was developed and used to characterize promoter strength, to verify suitability of bacterial gene modifications for expression in plant cells, and to express active antibody molecules. The system is based on suspension tobacco cells transformed by Agrobacterium in a transient way. Conditions such as pre‐culture of tobacco cells and the co‐cultivation period were identified as determinants to achieve high expression levels. Under established conditions the activity strength of CaMV (cauliflower mosaic virus) 35 S and ToMoTV (tomato mottle taino virus) AL1 promoters were compared. A modified cry gene sequence from Bacillus thuringiensis was expressed and detected by Western‐blot analysis. A monoclonal antibody against anti‐(hepatitis B virus surface antigen) was produced in such quantities as to allow testing of biological activity and preliminary characterization.

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