Plasmid transformation by electroporation of Leuconostoc paramesenteroides and its use in molecular cloning.

Applied and Environmental Microbiology
S DavidW M de Vos

Abstract

In this report, we demonstrate the utility of electroporation as an efficient method for genetic transformation of Leuconostoc paramesenteroides. We optimized several factors which determine the transformation frequency, resulting in transformation efficiencies of up to 4 x 10(3) transformants per micrograms of pNZ12 DNA, which contains the promiscuous Lactococcus lactis pSH71 replicon. Slightly lower efficiencies were obtained with a deletion derivative of the broad-host-range plasmid pAM beta 1. These plasmids could be stably maintained in L. paramesenteroides NZ6009 for more than 100 generations, even in the absence of selective pressure. In order to show the use of the developed host-vector system, we cloned the Lactococcus lactis gene encoding phospho-beta-galactosidase in L. paramesenteroides. Expression of this heterologous gene in L. paramesenteroides under control of Lactococcus lactis expression signals was evident from the presence, in transformants, of phospho-beta-galactosidase activity and a specific phospho-beta-galactosidase protein band on Western blots (immunoblots). In addition, we transformed a lactose-deficient derivative of L. paramesenteroides with a plasmid carrying a Lactococcus lactis-Escherichia coli ...Continue Reading

References

Nov 24, 1979·Nucleic Acids Research·H C Birnboim, J Doly
Oct 1, 1987·Applied and Environmental Microbiology·D van der Lelie, G Venema
Jul 11, 1988·Nucleic Acids Research·W J DowerC W Ragsdale
Feb 1, 1987·Applied and Environmental Microbiology·H J Tsai, W E Sandine
Apr 1, 1988·Analytical Biochemistry·S Fiedler, R Wirth
Apr 1, 1988·Biochimie·G A Somkuti, D H Steinberg
Feb 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·J F MillerL S Tompkins
Sep 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·M FrommV Walbot
Aug 1, 1984·Applied and Environmental Microbiology·J K Kondo, L L McKay
Nov 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·H PotterP Leder
Dec 1, 1984·Applied and Environmental Microbiology·P K Orberg, W E Sandine
Apr 1, 1980·Journal of Molecular Biology·M J Casadaban, S N Cohen
Mar 1, 1988·Applied and Environmental Microbiology·Ian B PowellBarrie E Davidson
May 1, 1961·Journal of Bacteriology·C Anagnostopoulos, J Spizizen

❮ Previous
Next ❯

Citations

Sep 1, 1990·FEMS Microbiology Reviews·L L McKay, K A Baldwin
Apr 9, 2008·Applied and Environmental Microbiology·Fang FangPaul W O'Toole
Oct 1, 1994·FEMS Microbiology Reviews·W M de Vos, E E Vaughan
Mar 27, 2001·Letters in Applied Microbiology·S M HoltK J Shin
Jun 9, 2004·Applied and Environmental Microbiology·Søren HelmarkPeter R Jensen
Jan 7, 2003·Applied and Environmental Microbiology·Fergal P RattrayDan Nilsson
Mar 5, 2005·Applied and Environmental Microbiology·Richard van KranenburgMichiel Kleerebezem
Jul 1, 1992·Journal of Bacteriology·S DavidW M de Vos
Jun 12, 2021·Applied Microbiology and Biotechnology·Robin DorauChristian Solem

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.