Open AccessA bifunctional Streptomyces‐E. coli promoter‐probe vetor
Author(s) -
Asturias Juan A.,
Liras Paloma,
Martin Juan F.
Publication year - 1990
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1990.tb04123.x
Subject(s) - multiple cloning site , pbr322 , plasmid , replicon , biology , streptomyces , genetics , promoter , terminator (solar) , cloning vector , gene , microbiology and biotechnology , molecular cloning , expression vector , recombinant dna , peptide sequence , bacteria , gene expression , ionosphere , physics , astronomy
A bifunctional Streptomyces‐E. coli promoter probe vector, pULJA30, has been developed to isolate and characterize nucleotide sequences involved in transcription initiation and regulation. The vector is derived from plasmid pIJ486 [1], carries the pIJ101 replicon and utilizes the promoterless amoniglycoside phosphotransferase ( neo ) as indicator gene. Important features of the new vector include: wide Streptomyces host range and as high a plasmid copy number as the parental pIJ486, an upstream transcriptional terminator ( t oop ) and a polylinker sequence with unique sites for Bam HI and Bgl II for flexible cloning, fragment re‐isolation and direct sequencing of promoter‐active inserts. pULJA30 also has an E. coli replicon (from pBR322) and the possibility of selection in Streptomycers and E. coli by using the tsr, neo and bla genes, which makes it very convenient to test the comparative functionality of Streptomyces promoters in E. coli .