An improved plasmid for the isolation and analysis of Streptomyces promoters.
A Streptomyces promoter-probe plasmid vector was developed to isolate and characterize nucleotide sequences involved in transcriptional regulation. This pCLL34 vector was derived from plasmid pARC1 carrying the SLP1.2 origin of replication [Horinouchi and Beppu, J. Bacteriol. 162 (1985) 406-412]. Important features of the new vector include moderate host range, low copy number, simple identification of promoter activity by brown pigment production, an upstream transcriptional terminator, and a polylinker sequence containing a unique BamHI site for flexible cloning and fragment reisolation. The vector also provides the capability to directly determine the sequence of promoter-active inserts by dideoxy chain-termination sequencing methods using double-stranded plasmid templates and the appropriate oligodeoxynucleotide primers.
Feitelson
J S
JS
Department of Microbial Genetics, Lederle Laboratories, American Cyanamid Company, Pearl River, NY 10965.
D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular / Q000379:methods*; D004262:DNA Restriction Enzymes; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D011401:Promoter Regions, Genetic; D013302:Streptomyces / Q000235:genetics*; D013728:Terminator Regions, Genetic
D009838:Oligodeoxyribonucleotides; D004262:DNA Restriction Enzymes