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Title: A geminivirus AYVV-derived shuttle vector for tobacco BY2 cells
Authors: Tamilselvi, D.
Anand, G. 
Swarup, S. 
Keywords: Ageratum yellow vein virus
Foreign gene expression
Shuttle vector
Issue Date: Aug-2004
Citation: Tamilselvi, D.,Anand, G.,Swarup, S. (2004-08). A geminivirus AYVV-derived shuttle vector for tobacco BY2 cells. Plant Cell Reports 23 (1-2) : 81-90. ScholarBank@NUS Repository.
Abstract: We have developed a plant-Escherichia coli pASV shuttle vector from the essential elements of the Ageratum yellow vein virus (AYVV). The geminivirus vector contains the AYVV genome with the coat-protein deletion, the E. coli vector backbone of pUC19, a unique cloning site and gene expression cassettes for plant selection and reporter gene activity. The replication of pASV vectors was compared in Nicotiana benthamiana and N. tabacum BY2 cells, and the latter were found to be suitable for long-term maintenance of the vectors in culture. The vector DNA was detected at regular intervals by PCR, β-glucuronidase expression analysis and plasmid rescue during a 4-month culture period. A novel methylation-based PCR assay was carried out to show de novo replication for pASV-derived vectors in 2-month-old tobacco BY2 cell lines. This is the first report of the extrachromosomal replication of monopartite begomovirus with stability and foreign gene expression in long-term cell cultures.
Source Title: Plant Cell Reports
ISSN: 07217714
Appears in Collections:Staff Publications

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