Cloning of circular DNAs from microorganisms using a novel plasmid capture system.
Plasmid capture system (PCS) facilitates cloning and manipulation of circular double-stranded DNA. We recently developed an improved PCS (PCS-LZ) to clone relatively large DNA molecules of 30-150 kb. The PCS-LZ donor consists of a mini-F replicon and a kanamycin resistance marker between Tn7 left and Tn7 right ends. Both the replicon and marker gene of the PCS-LZ donor are transferred into target plasmid DNAs by in vitro transposition, followed by replication in E. coli. Colonies are tested for lacZ expression by blue/white screening. Circular DNAs were obtained from plasmids of Bacillus thuringiensis, genome segments of Cotesia glomerata bracovirus and polymorphic genomes of Autographa californica nucleopolyhedrovirus. PCS-LZ is a powerful tool for use in genomic analysis and mutagenesis in microorganisms including invertebrate pathogens.
Roh
Jong Yul
JY
Department of Agricultural Biotechnology, Seoul National University, Republic of Korea.
Wang
Yong
Y
Liu
Qin
Q
Tao
Xueying
X
Choi
Jae Young
JY
Shim
Hee Jin
HJ
Xu
Hong Guang
HG
Lee
Seungdon
S
Woo
Soo Dong
SD
Jin
Byung Rae
BR
Je
Yeon Ho
YH
D001413:Bacillus thuringiensis / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D004270:DNA, Circular / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D008954:Models, Biological; D017924:Nucleopolyhedroviruses / Q000235:genetics; D010957:Plasmids / Q000235:genetics*
D004270:DNA, Circular; D004279:DNA, Viral