Construction and characterization of bacterial artificial chromosome libraries from the silkworm, Bombyx mori.
Two bacterial artificial chromosome (BAC) libraries were constructed using nuclear DNA from posterior silkglands of the silkworm (Bombyx mori) strains p50 and C108. The libraries contain a total of 36,864 clones, or approximately 9 genome equivalents. The average insert sizes in the libraries were 134.5 kb and 120.8 kb, respectively. PCR-based screening was performed on the p50 library using probes for 34 sequence-tagged sites (STSs). Between 3 and 11 (6.1 hits on average) clones were isolated with each STS, in good agreement with the library size, 5.8 genome equivalents. The previously reported close linkage between the Bombyx homologs of the invected (Bm in) and engrailed (Bm en) genes was confirmed by construction of a BAC contig that contained both. Moreover, screening revealed novel information about the chromosomal organization of the sericin-1 and DH-PBAN genes, which were localized within a 22-kb interval and are divergently oriented. These results show that it is possible to construct contigs and analyze chromosome organization using these libraries.
National Institute of Sericultural and Entomological Science, Tsukuba, Ibaraki, Japan.
C
C
Wu
S
S
Asakawa
N
N
Shimizu
S
S
Kawasaki
Y
Y
Yasukochi
D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000235:genetics* / Q000254:growth & development; D002876:Chromosomes, Bacterial; D017931:DNA Primers; D004791:Enzyme Inhibitors; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D005822:Genetic Vectors; D019476:Insect Proteins / Q000235:genetics*; D007814:Larva; D009479:Neuropeptides / Q000235:genetics; D010456:Peptides, Cyclic / Q000235:genetics; D016133:Polymerase Chain Reaction; D011679:Pupa; D016324:Sequence Tagged Sites; D047030:Sericins; D014170:Transformation, Genetic
D017931:DNA Primers; D004791:Enzyme Inhibitors; D019476:Insect Proteins; D009479:Neuropeptides; D010456:Peptides, Cyclic; D047030:Sericins; C060896:pheromone-biosynthesis-activating neuropeptide I