The construction and characteristics of a BAC library for Cucumis sativus L. 'B10'.
Cloning using bacterial artificial chromosomes (BACs) can yield high quality genomic libraries, which are used for the physical mapping, identification and isolation of genes, and for gene sequencing. A BAC genomic library was constructed from high molecular weight DNA (HMW DNA) obtained from nuclei of the cucumber (Cucumis sativus L. cv. Borszczagowski; B10 line). The DNA was digested with the HindIII restriction enzyme and ligated into the pCC1BAC vector. The library consists of 34,560 BAC clones with an average insert size of 135 kb, and 12.7x genome coverage. Screening the library for chloroplast and mitochondrial DNA content indicated an exceptionally low 0.26% contamination with chloroplast DNA and 0.3% with mitochondrial DNA.
Gutman
Wojciech
W
Department of Biochemistry, Warsaw Agricultural University, 02-776, Warsaw, Poland. wojciech_gutman@sggw.pl
Pawełkowicz
Magdalena
M
Woycicki
Rafał
R
Piszczek
Ewa
E
Przybecki
Zbigniew
Z
D002874:Chromosome Mapping; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D018553:Cucumis sativus / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D005822:Genetic Vectors; D008970:Molecular Weight; D018515:Plant Leaves / Q000235:genetics; D014169:Transformation, Bacterial
D018744:DNA, Plant