Plant & Animal Genome V Conference
Town & Country Hotel, San Diego, CA, January 12-16, 1997.
PAG-V: S11 - A BINARY-BAC SYSTEM FOR PLANT TRANSFORMATION WITH HIGH MOLECULAR WEIGHT DNA
S11
A BINARY-BAC SYSTEM FOR PLANT TRANSFORMATION WITH HIGH MOLECULAR WEIGHT DNA
HAMILTON, CAROL M.(1), Anne Frary(1), Candice Lewis(2), Steven D. Tanksley(2)
1. Plant Science Center, Center for Advanced Technology/Biotechnology
2. Department of Plant Breeding and Biometry
The binary- bacterial artificial chromosome (BIBAC) vector is based on the binary vector for Agrobacterium-mediated
transformation of plants and the bacterial artificial chromosome (BAC) vector for the construction of genomic DNA libraries with large
(100-300 kb) inserts in Escherichia coli. The BIBAC is capable of transferring high molecular weight DNA, intact, into plant
chromosomes and is suitable for the construction of plant genomic DNA libraries. Special virulence helper plasmids are required to effect
Agrobacterium-mediated transformation of plants with high molecular weight DNA fragments. The helper plasmids are compatible
with and are present in addition to the BIBAC in the A. tumefaciens host. When used in combination with virulence- enhanced A.
tumefaciens strains, the BIBAC vector is capable of transferring at least 150 kb of foreign DNA into the plant nuclear genome. The
transferred DNA is faithfully inherited in the progeny. The BIBAC system was first demonstrated in tobacco, and has recently been established
for tomato. Factors contributing to the efficiency and integrity of high molecular weight DNA transfer events will be discussed. The ability to
introduce high molecular weight DNA into plant chromosomes should accelerate gene identification, facilitate identification of quantitative
trait loci, and will create new possibilities for studies of gene expression and chromosome organization.