title: Construction of new cloning vectors for Brevibacterium lactofermentum.
abstract: Two plasmid cloning vectors (pULMJ55 and pULMJ95) were constructed for Brevibacterium lactofermentum using the origin of replication of the endogenous plasmid pBL1. Plasmid pULMJ55 is a replacement vector with transcriptional terminators from the B. lactofermentum trp operon flanking the BglII cloning sites. Religation of the BglII digested vector without insert creates a 376 bp perfect palindrome that is not tolerated in B. lactofermentum, giving positive selection for recombinant plasmids with inserts. Plasmid pULMJ95 contains the promoter-less alpha-amylase gene from Streptomyces griseus downstream of the trp terminator and is particularly suitable for the detection of promoters which are activated late during the growth phase. alpha-Amylase is secreted and its activity can be detected using simple plate tests.
authors.affiliation: Departamento de Ecología, Genética y Microbiología, Universidad de León, Spain.
authors.forename: R F
authors.initials: RF
authors.lastname: Cadenas
authors.forename: C
authors.initials: C
authors.lastname: Fernández-González
authors.forename: J F
authors.initials: JF
authors.lastname: Martín
authors.forename: J A
authors.initials: JA
authors.lastname: Gil
mesh_terms: D001951:Brevibacterium / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D003352:Corynebacterium / Q000235:genetics; D005798:Genes, Bacterial; D017930:Genes, Reporter; D005822:Genetic Vectors; D015335:Molecular Probes; D009876:Operon; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D018741:Replication Origin; D013305:Streptomyces griseus / Q000201:enzymology / Q000235:genetics; D014364:Tryptophan / Q000235:genetics; D000516:alpha-Amylases / Q000235:genetics
chemical_list: D015335:Molecular Probes; D014364:Tryptophan; D000516:alpha-Amylases