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Transient Expression System (transient + expression_system)
Selected AbstractsIn vitro Transient Expression System of Latex C-serum was used for Analysis of Hevein Promoter in Response to Abscisic Acid in Hevea brasiliensisJOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 3 2008Xiao-Wen Fei Abstract Hevein has been found to be an essential element in coagulation of rubber particles in latex of rubber trees. In a previous study, we cloned a 1 241-bp fragment of a 5, upstream region of the hevein gene by genome walking. This fragment was analyzed by a 5, end nested deletion method in the present study, fused with a uidA (gus) gene to produce a series of tested constructs, which were transferred into C-serum of latex and the Gus activities were detected. Results showed that the fragment from ,749 to ,292 was sufficient for expression of gus gene in latex, and the fragment from ,292 to ,168 was crucial in response to abscisic acid inducement. In a transient transgenic test of rubber leaf with particle bombardment, construct Hev749 conferred gus -specific expression in veins, in which the latex tubes mainly distributed. This implies that the fragment from ,749 to ,292 was laticiferous-specific. [source] A transient expression vector for recombinant protein production in Chinese hamster ovary cellsJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 1 2006Mimi ML Liao Abstract An expression vector was specifically designed for use in Chinese hamster ovary (CHO) cells to enhance the level of protein production in a transient expression system. Two key components that can increase protein production transiently are the promoter used to drive recombinant gene expression and the template copy number. In this study the modified and metal-inducible metallothionein (M2.6) promoter was shown to be superior to the human cytomegalovirus (CMV) and to the simian virus SV40 promoters. Plasmid replication was achieved using the Polyoma (Py) virus origin of replication (PyOri) and the Py Large T antigen (PyLT). An expression vector containing Py elements was shown to replicate extensively in CHO cells. The combination of the metal-inducible M2.6 promoter and episomal replication of the expression vector, named pPyOriLT resulted in elevated levels of transgene expression following transient transfection of CHO cells Copyright © 2005 Society of Chemical Industry [source] Transient expression of a vacuolar peroxidase increases susceptibility of epidermal barley cells to powdery mildewMOLECULAR PLANT PATHOLOGY, Issue 6 2001Brian Kåre Kristensen summary The expression of genes encoding the peroxidases, Prx7 and Prx8, is induced in barley leaf tissue after inoculation with the barley powdery mildew fungus, Blumeria graminis f.sp. hordei (DC) Speer (Bgh). The role of these peroxidases in general barley defence responses against fungal attack was investigated using a transient expression system. Colonization frequencies of Bgh on cells transfected with Prx7 or Prx8 expression-, mutant- or fusion-DNA constructs were compared to the frequencies on cells expressing a ,-glucuronidase (GUS) control construct. Twice the number of powdery mildew colonies were observed on cells expressing Prx7 as compared to control cells. Introduction of either mutant or truncated versions of Prx7 showed that decreased resistance against Bgh was dependent on the presence of the C-terminal signal peptide required for correct subcellular targeting, but not affected significantly by mutations in the catalytic centre. No impact on Bgh performance was observed after the introduction of Prx8 or mutant constructs. An enhanced accumulation of the apoplastic Prx8 was verified by immunocytology. These results indicate a more complex role of peroxidases in defence responses than was previously suspected. [source] Manufacturing antibodies in the plant cellBIOTECHNOLOGY JOURNAL, Issue 12 2009Diego Orzáez Dr. Abstract Plants have long been considered advantageous platforms for large-scale production of antibodies due to their low cost, scalability, and the low chances of pathogen contamination. Much effort has therefore been devoted to efficiently producing mAbs (from nanobodies to secretory antibodies) in plant cells. Several technical difficulties have been encountered and are being overcome. Improvements in production levels have been achieved by manipulation of gene expression and, more efficiently, of cell targeting and protein folding and assembly. Differences in mAb glycosylation patterns between animal and plant cells are being successfully addressed by the elimination and introduction of the appropriate enzyme activities in plant cells. Another relevant battlefield is the dichotomy between production capacity and speed. Classically, stably transformed plant lines have been proposed for large scale mAb production, whereas the use of transient expression systems has always provided production speed at the cost of scalability. However, recent advances in transient expression techniques have brought impressive yield improvements, turning speed and scalability into highly compatible assets. In the era of personalized medicines, the combination of yield and speed, and the advances in glyco-engineering have made the plant cell a serious contender in the field of recombinant antibody production. [source] | ||||||||||