PMID: 9544937Apr 17, 1998Paper

Development of an improved cloning vector and transformation system in Amycolatopsis mediterranei (Nocardia mediterranei)

The Journal of Antibiotics
Rup LalS Lal

Abstract

A versatile plasmid cloning vector pRL60 carrying kanamycin/neomycin resistance (km/neo), erythromycin resistance (ermE) and alpha-amylase (alpha-amy) marker genes that replicates in various Amycolatopsis mediterranei strains and Escherichia coli has been constructed. This cloning vector has been derived from a hybrid plasmid pRL50, which was developed by cloning ermE from pIJ4026 into a pRL1 derivative pULAM2. While cloning ermE into the BamHI site of pULAM2, only a hybrid plasmid pRL50 with an additional copy of pULAM2 was selected. Thus pRL50 (18.7 kb) contained two copies each of the km/neo, alpha-amy, and one copy of ermE. When pRL50 was transformed into A. mediterranei DSM 40773 through electroporation and selected under erythromycin resistance, the plasmid underwent a spontaneous deletion of 8.5 kb fragment resulting in the formation of plasmid pRL60. pRL60 (10.2 kb) is a shuttle vector between A. mediterranei and E. coli with three marker genes: km/neo, ermE and alpha-amy. ermE is expressed in A. mediterranei thus allowing good selection of transformants. The alpha-amy gene of pRL60 is also expressed in A. mediterranei DSM 40773 and its activity can be easily detected on starch containing medium after iodine staining. M...Continue Reading

Citations

Jul 8, 2000·Letters in Applied Microbiology·P S Murali KrishnaL Venkateswar Rao
Apr 11, 2003·Journal of Industrial Microbiology & Biotechnology·Gauri DhingraRup Lal
Oct 17, 2014·Indian Journal of Microbiology·Anjali SaxenaRup Lal
Dec 4, 2016·Applied and Environmental Microbiology·Florian MeyerAlexander Steinbüchel
Nov 21, 2017·Indian Journal of Microbiology·Priya SinghRup Lal
Mar 1, 2007·Indian Journal of Microbiology·S Malhotra, R Lal
Apr 20, 2014·Applied Microbiology and Biotechnology·Christian Fleige, Alexander Steinbüchel
Jul 29, 2004·International Journal of Systematic and Evolutionary Microbiology·Shashi BalaRup Lal
May 7, 2004·Organic Letters·Jiantao Guo, J W Frost

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