Journal Article

T-strand integration in maize protoplasts after codelivery of a T-DNA substrate and virulence genes

Hansen, G, Chilton, M.D, Shillito, R.D

1997

Overview

We describe a plant protoplast transformation method that provides transformants with a simple pattern of integration of a foreign gene. The approach is to deliver into plant protoplasts by direct gene transfer the Agrobacterium virulence genes virD1 and virD2 with or without virE2, together with a target plasmid containing a gene of interest flanked by Agrobacterium T-DNA border repeat sequences of 25 bp. We present evidence of T-DNA formation in maize protoplasts and its integration into the maize genome. The frequency of VirD1-VirD2-mediated integration events was about 20-35% of the total number of transformants. The addition of virE2 doubled the transformation efficiency. The method described here is of sufficient efficiency and simplicity to be useful for the production of transgenic plants with single-copy well-defined transgenic inserts

• Publisher: National Academy of Sciences of the United States of America,National Acad Sciences,National Academy of Sciences,The National Academy of Sciences of the USA
• Subject: ADN / Agrobacterium / Agrobacterium radiobacter / AGROBACTERIUM TUMEFACIENS / ANTIBIOTICOS / ANTIBIOTICS / ANTIBIOTIQUE / Bacterial Proteins / Bacterial Proteins - biosynthesis / Bacterial Proteins - genetics / Base Sequence / Biological Sciences / biosynthesis / Botany / CAL / CALLO / CALLUS / Corn / Deoxyribonucleic acid / DIRECT DNA UPTAKE / DNA / DNA probes / DNA, Bacterial / DNA, Bacterial - genetics / DNA, Bacterial - metabolism / DNA, Single-Stranded / DNA, Single-Stranded - genetics / DNA, Single-Stranded - metabolism / drug resistance / DRUGS / EXPRESION GENICA / EXPRESSION DES GENES / GENE / GENE EXPRESSION / GENE TRANSFER / Gene Transfer Techniques / GENES / GENETIC TRANSFORMATION / genetics / Genomes / INTEGRATION INSERTION JUNCTIONS / KINASES / MEDICAMENT / MEDICAMENTOS / metabolism / microbiology / Molecular Sequence Data / NEOMYCIN PHOSPHOTRANSFERASE II / NUCLEOTIDE SEQUENCE / nucleotide sequences / PAROMOMYCIN / PATHOGENICITY / PATOGENICIDAD / phosphotransferases (kinases) / Plants / PLASMID VECTORS / Plasmids / POUVOIR PATHOGENE / PROTOPLASTE / PROTOPLASTOS / PROTOPLASTS / Repetitive Sequences, Nucleic Acid / REPORTER GENES / RESISTANCE AUX PRODUITS CHIMIQUES / RESISTANCE TO CHEMICALS / RESISTENCIA A PRODUCTOS QUIMICOS / Rhizobium / Rhizobium - genetics / Rhizobium - pathogenicity / SECUENCIA NUCLEOTIDICA / SEQUENCE NUCLEOTIDIQUE / TRANSFERASAS / TRANSFERASE / TRANSFERASES / TRANSFERENCIA DE GENES / TRANSFERT DE GENE / TRANSFORMACION GENETICA / TRANSFORMATION GENETIQUE / VIRD1 GENE / VIRD2 GENE / Virulence / Virulence Factors / ZEA MAYS / Zea mays - genetics / Zea mays - microbiology