High-throughput cloning of Campylobacter jejuni ORfs by in vivo recombination in Escherichia coli.
A rate-limiting and costly step in many proteomics analyses is the cloning of all of the ORFs for an organism into technique-specific vectors. Here, we describe the generation of a Campylobacter jejuni expression clone set using a high-throughput cloning approach based on recombination in E. coli. The approach uses native E. coli recombination functions and requires no in vitro enzymatic steps or special strains. Our results indicate that this approach is an efficient and economical alternative for high-throughput cloning.
Parrish
Jodi R
JR
Center for Molecular Medicine and Genetics, Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, 540 East Canfield Ave, Detroit, Michigan 48201, USA.
Limjindaporn
Thawornchai
T
Hines
Julie A
JA
Liu
Jiayou
J
Liu
Guozhen
G
Finley
Russell L
RL
D016123:Campylobacter jejuni / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D011995:Recombination, Genetic