BIOMAT Database Logo BIOMAT Database Logo

High-frequency transformation of Brevibacterium lactofermentum protoplasts by plasmid DNA. 1985

R I Santamaria, and J A Gil, and J F Martin

An efficient polyethylene glycol-assisted method for transformation of Brevibacterium lactofermentum protoplasts that uses plasmid vectors has been developed. Two small plasmids, pUL330 (5.2 kilobases) and pUL340 (5.8 kilobases), both containing the kanamycin resistance gene from transposon Tn5 and the replication origin of the natural plasmid pBL1 of B. lactofermentum, were selected as vectors. Supercoiled forms of the plasmids yielded a 100-fold higher transformation frequency than did linear forms. The optimal transformation frequency was achieved with 10 ng of DNA in 1 ml of transformation buffer. Higher concentrations of plasmid DNA resulted in a decrease in transformation frequency per microgram of DNA. Optimal transformation was obtained with 25 to 35% polyethylene glycol 6000. Under optimal conditions, 10(6) transformants per microgram of DNA were obtained.

UI MeSH Term Description Entries
D008274 Magnesium A metallic element that has the atomic symbol Mg, atomic number 12, and atomic weight 24.31. It is important for the activity of many enzymes, especially those involved in OXIDATIVE PHOSPHORYLATION.
D009113 Muramidase A basic enzyme that is present in saliva, tears, egg white, and many animal fluids. It functions as an antibacterial agent. The enzyme catalyzes the hydrolysis of 1,4-beta-linkages between N-acetylmuramic acid and N-acetyl-D-glucosamine residues in peptidoglycan and between N-acetyl-D-glucosamine residues in chitodextrin. EC 3.2.1.17. Lysozyme,Leftose,N-Acetylmuramide Glycanhydrolase,Glycanhydrolase, N-Acetylmuramide,N Acetylmuramide Glycanhydrolase
D010957 Plasmids Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS. Episomes,Episome,Plasmid
D011092 Polyethylene Glycols Polymers of ETHYLENE OXIDE and water, and their ethers. They vary in consistency from liquid to solid depending on the molecular weight indicated by a number following the name. They are used as SURFACTANTS, dispersing agents, solvents, ointment and suppository bases, vehicles, and tablet excipients. Some specific groups are NONOXYNOLS, OCTOXYNOLS, and POLOXAMERS. Macrogols,Polyoxyethylenes,Carbowax,Macrogol,Polyethylene Glycol,Polyethylene Oxide,Polyethyleneoxide,Polyglycol,Glycol, Polyethylene,Glycols, Polyethylene,Oxide, Polyethylene,Oxides, Polyethylene,Polyethylene Oxides,Polyethyleneoxides,Polyglycols,Polyoxyethylene
D011523 Protoplasts The protoplasm and plasma membrane of plant, fungal, bacterial or archaeon cells without the CELL WALL. Protoplast
D001951 Brevibacterium A gram-positive organism found in dairy products, fresh and salt water, marine organisms, insects, and decaying organic matter.
D002118 Calcium A basic element found in nearly all tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes. Coagulation Factor IV,Factor IV,Blood Coagulation Factor IV,Calcium-40,Calcium 40,Factor IV, Coagulation
D013696 Temperature The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms. Temperatures
D014169 Transformation, Bacterial The heritable modification of the properties of a competent bacterium by naked DNA from another source. The uptake of naked DNA is a naturally occuring phenomenon in some bacteria. It is often used as a GENE TRANSFER TECHNIQUE. Bacterial Transformation

Related Publications

R I Santamaria, and J A Gil, and J F Martin
High frequency transformation of Streptomyces niveus protoplasts by plasmid DNA.
November 1991, The Journal of applied bacteriology,
R I Santamaria, and J A Gil, and J F Martin
High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA.
January 1979, Molecular & general genetics : MGG,
R I Santamaria, and J A Gil, and J F Martin
Transfer of plasmid DNA to Brevibacterium lactofermentum by electrotransformation.
October 1990, Journal of general microbiology,
R I Santamaria, and J A Gil, and J F Martin
High-efficiency transformation of Streptococcus lactis protoplasts by plasmid DNA.
August 1986, Applied and environmental microbiology,
R I Santamaria, and J A Gil, and J F Martin
An efficient method for the introduction of viral DNA into Brevibacterium lactofermentum protoplasts.
June 1986, Journal of general microbiology,
R I Santamaria, and J A Gil, and J F Martin
Transformation of Streptococcus bovis protoplasts by plasmid DNA.
February 1996, Letters in applied microbiology,
R I Santamaria, and J A Gil, and J F Martin
Transformation of Streptococcus lactis Protoplasts by Plasmid DNA.
May 1982, Applied and environmental microbiology,
R I Santamaria, and J A Gil, and J F Martin
Complete nucleotide sequence of a native plasmid of Brevibacterium lactofermentum.
June 1986, Nucleic acids research,
R I Santamaria, and J A Gil, and J F Martin
Transformation of plasmid DNA into Streptomyces at high frequency.
July 1978, Nature,
R I Santamaria, and J A Gil, and J F Martin
[Nucleotide sequence and structural analysis of cryptic plasmid pBL90 from Brevibacterium lactofermentum].
November 2016, Genetika,
Copied contents to your clipboard!