This is the first report of large insert genomic DNA libraries constructed
in a binary-BAC (BIBAC) vector. Genomic DNA libraries containing approximat
ely 4.6 haploid nuclear genomic equivalents were constructed for Lycopersic
on esculentum (cv. Mogeor) and Lycopersicon pennellii (LA716) in the BIBAC2
vector. The L. esculentum library has an average insert size of 125 kb and
is comprised of 42 272 individual colonies stored as frozen cultures in a
384-well format (108 plates). The L. pennellii library has an average inser
t size of 90 kb and is comprised of 53 760 individual clones (140 384-well
plates). In each of the libraries, it is estimated that 90% of the colonies
contain genomic DNA inserts. The composition of the L. esculentum and L. p
ennellii libraries was determined by analyzing a series of randomly selecte
d clones. The L. esculentum library was surveyed for clones containing chlo
roplast DNA (1.4%), mitochondrial DNA (0.012%) and repetitive DNA motifs. B
IBAC clones that may contain a gene of interest can be identified from thes
e libraries by colony hybridization with homologous or heterologous probes
or by PCR pooling techniques. Once identified, BIBAC genomic DNA library cl
ones are immediately suitable for Agrobacterium tumefaciens-mediated plant
transformation.