Lorist2, a cosmid with transcriptional terminators insulating vector genes from interference by promoters within the insert: effect on DNA yield and cloned insert frequency.
Transcription terminators have been included in a phage-lambda-replicon-based cosmid vector, Lorist2, to insulate vector genes against transcriptional interference from cloned insert DNA. DNA yields of recombinant clones containing Escherichia coli genomic DNA inserts are more even for Lorist2 than with its progenitor LoristB. However, the terminators provide only a partial reduction in the over-representation of r X DNA-containing clones generally observed in cosmid libraries of Caenorhabditis elegans DNA, suggesting that causes other than transcriptional readthrough into the vector contribute to this problem.
Gibson
T J
TJ
Coulson
A R
AR
Sulston
J E
JE
Little
P F
PF
D000818:Animals; D002107:Caenorhabditis / Q000235:genetics; D002455:Cell Division; D003001:Cloning, Molecular / Q000379:methods*; D003360:Cosmids; D004261:DNA Replication; D004926:Escherichia coli / Q000235:genetics; D005809:Genes, Regulator; D005822:Genetic Vectors; D011401:Promoter Regions, Genetic; D013728:Terminator Regions, Genetic