TY - JOUR
T1 - DNA shuttling between plasmid vectors and a genome vector
T2 - Systematic conversion and preservation of DNA libraries using the Bacillus subtilis genome (BGM) vector
AU - Kaneko, Shinya
AU - Akioka, Manami
AU - Tsuge, Kenji
AU - Itaya, Mitsuhiro
PY - 2005/6/24
Y1 - 2005/6/24
N2 - The combined use of the contemporary vector systems, the bacterial artificial chromosome (BAC) vector and the Bacillus subtilis genome (BGM) vector, makes possible the handling of giant-length DNA (above 100 kb). Our newly constructed BGM vector efficiently integrated DNA prepared in the BAC vector. A BAC library comprised of 18 independent clones prepared from mitochondrial DNA (mtDNA) of Arabidopsis thaliana was converted to a parallel BGM library using the new BGM vector. The effectiveness of the combined use of the vector systems was confirmed by the stable recovery of all 18 DNAs as BAC clones from the respective BGM clones. We show that DNA in BGM was stably preserved at room temperature after spore formation of the host B. subtilis. Rapid and stable shuttling between Escherichia coli and the B. subtilis host, combined with spore-mediated DNA storage, may facilitate the long-term and low-cost preservation and the transportation of DNA resources.
AB - The combined use of the contemporary vector systems, the bacterial artificial chromosome (BAC) vector and the Bacillus subtilis genome (BGM) vector, makes possible the handling of giant-length DNA (above 100 kb). Our newly constructed BGM vector efficiently integrated DNA prepared in the BAC vector. A BAC library comprised of 18 independent clones prepared from mitochondrial DNA (mtDNA) of Arabidopsis thaliana was converted to a parallel BGM library using the new BGM vector. The effectiveness of the combined use of the vector systems was confirmed by the stable recovery of all 18 DNAs as BAC clones from the respective BGM clones. We show that DNA in BGM was stably preserved at room temperature after spore formation of the host B. subtilis. Rapid and stable shuttling between Escherichia coli and the B. subtilis host, combined with spore-mediated DNA storage, may facilitate the long-term and low-cost preservation and the transportation of DNA resources.
KW - Genome vector
KW - Recombination
KW - Recombinational transfer
KW - Spore formation
KW - Transformation
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U2 - 10.1016/j.jmb.2005.04.041
DO - 10.1016/j.jmb.2005.04.041
M3 - Article
C2 - 15913652
AN - SCOPUS:20344385510
SN - 0022-2836
VL - 349
SP - 1036
EP - 1044
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 5
ER -