RRC ID 61163
Author Itaya M, Tsuge K, Koizumi M, Fujita K.
Title Combining two genomes in one cell: stable cloning of the Synechocystis PCC6803 genome in the Bacillus subtilis 168 genome.
Journal Proc Natl Acad Sci U S A
Abstract Cloning the whole 3.5-megabase (Mb) genome of the photosynthetic bacterium Synechocystis PCC6803 into the 4.2-Mb genome of the mesophilic bacterium Bacillus subtilis 168 resulted in a 7.7-Mb composite genome. We succeeded in such unprecedented large-size cloning by progressively assembling and editing contiguous DNA regions that cover the entire Synechocystis genome. The strain containing the two sets of genome grew only in the B. subtilis culture medium where all of the cloning procedures were carried out. The high structural stability of the cloned Synechocystis genome was closely associated with the symmetry of the bacterial genome structure of the DNA replication origin (oriC) and its termination (terC) and the exclusivity of Synechocystis ribosomal RNA operon genes (rrnA and rrnB). Given the significant diversity in genome structure observed upon horizontal DNA transfer in nature, our stable laboratory-generated composite genome raised fundamental questions concerning two complete genomes in one cell. Our megasize DNA cloning method, designated megacloning, may be generally applicable to other genomes or genome loci of free-living organisms.
Volume 102(44)
Pages 15971-6
Published 2005-11-1
DOI 10.1073/pnas.0503868102
PII 0503868102
PMID 16236728
PMC PMC1276048
MeSH Bacillus subtilis / genetics* Cloning, Molecular / methods* Contig Mapping Genetic Vectors Genome, Bacterial* Plasmids / genetics Replication Origin Synechocystis / genetics* rRNA Operon / genetics
IF 9.58
Resource
Prokaryotes B. subtilis MBS902 MBS903 MBS904 MBS905 MBS906 MBS907 MBS908 MBS909 MBS910 MBS911 MBS912 MBS913 MBS914 MBS915 MBS916 MBS917 MBS918 MBS919 MBS920 MBS921 MBS922 MBS923 MBS924 MBS925 MBS926 MBS927 MBS928 MBS929 MBS930 MBS931 MBS932 MBS933 MBS934 MBS935 MBS936