Vectors for Cloning in Plant Cells

R. Deblaere, A. Reynaerts, H. Höfte, J. P. Hernalsteens, J. Leemans, M. Van Montagu

Onderzoeksoutput: Articlepeer review

114 Citaten (Scopus)

Samenvatting

This chapter discusses vectors for cloning in plant cells. The chapter presents recent octopine Ti plasmid-derived vectors and their use to transfer and express foreign genes in plants. Plant cells transformed with this avirulent tDNA retain their intrinsic capacity to redifferentiate into complete plants. Moreover, the tDNA vectors contain a dominant selectable marker gene, mainly a chimeric neomycin phosphotransferase gene that provides the transformed plant cell with a selectable kanamycin resistance trait. pGV2260 contains the intact oir region, which encodes Ti plasmid functions necessary for tDNA transfer and/or integration. These functions can act in trans to the tDNA. The chapter discusses cointegration vectors and binary vectors. The chapter includes the steps to introduce a foreign gene into a Ti plasmid vector. The chapter also discusses the expression of chimeric genes in plant cells, plant transformation, the transformation of tobacco leaf fragments, the analysis of gene expression in transformed plants, neomycin phosphotransferase assay, and tDNA organization in transformed plants.

Originele taal-2English
Pagina's (van-tot)277-292
Aantal pagina's16
TijdschriftMethods in Enzymology
Volume153
Nummer van het tijdschriftC
DOI's
StatusPublished - 1 jan 1987

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