Open AccessConstruction of new cloning vectors for Brevibacterium lactofermentum
Author(s) -
Cadenas Rosa F.,
FernándezGonzález Cristina,
Martín Juan F.,
Gil JoséA.
Publication year - 1996
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.1574-6968.1996.tb08083.x
Subject(s) - plasmid , bglii , multiple cloning site , biology , cloning vector , shuttle vector , brevibacterium , operon , cloning (programming) , periplasmic space , molecular cloning , genetics , microbiology and biotechnology , gene , recombinant dna , vector (molecular biology) , escherichia coli , ecori , bacteria , gene expression , computer science , programming language , microorganism
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 Bgl II cloning sites. Religation of the Bgl II 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 α‐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. α‐Amylase is secreted and its activity can be detected using simple plate tests.