Relevance of Endotoxin Contamination in Plasmid DNA
نویسندگان
چکیده
Endotoxins are known to be frequent contaminants in plasmid DNA extracted from bacteria. While many typical downstream applications such as sequencing and cloning are not affected by the presence of these molecules, endotoxins are of great concern to researchers in the pharmaceutical and biotechnology industries, since they may serve as potent inhibitors to more sensitive downstream applications such as the transfection of sensitive eukaryotic cells.
منابع مشابه
Endotoxin Contamination of Large Volume Parenterals as Detected by Limulus Amebocyte Lysate as an Alternative to Rabbit Pyrogen Test
The possibility of replacing the rabbit pyrogen test by the Limulus Amebocyte lysate (LAL) test, as a final release test for Large Volume Parenterals (LVPs) was investigated. The sensitivity and specificity of the LAL test and rabbit pyrogen assay were studied by means of artificially contaminated parenterals. Various LVPs (mostly containing electrolytes) were spiked with 0.5 and 5 EUlml o...
متن کاملDirect in vivo transfer of plasmid DNA into murine tumors: effects of endotoxin presence and transgene localization.
The purpose of this study was to investigate the effect of endotoxin presence in plasmid DNA preparations on the efficiency of transfection achieved in vivo with B16(F10) and Renca tumors and to determine transgene localization. Our data show that endotoxin markedly decreases the efficiency of transfection. Furthermore, the transgene transferred in vivo can be found in both neoplastic and norma...
متن کاملConstruction of Mtb72F Plasmid as a DNA Vaccine Candidate for Mycobacterium tuberculosis
Background: With one-third of the world’s population infected, tuberculosis (TB) is one of the most common infectious diseases and a major public health problem, especially in developing countries. The efficacy of the BCG vaccine for controlling the disease in adults is poor. The development of an effective TB vaccine is a global objective. An effective tuberculosis vaccine should s...
متن کاملConstruction of a Synthetic Vector for Preparation of a 100 Base Pair DNA Ladder
DNA size markers are widely used to estimate the size of DNA samples on agarose or polyacrylamide gelelectrophoresis (PAGE). DNA markers can be prepared by mixing PCR products with definite sizes.Alternatively, they are prepared by restriction enzyme digestion of the genomic DNA of bacteriophages ornatural and synthetic DNA plasmids. The present study describes engineering of ...
متن کاملConstruction of a Eukaryotic Plasmid Encoding Bacillus anthracis Protective Antigen, a Candidate for DNA Vaccine
Background: DNA immunization with plasmid DNA encoding bacterial, viral, parasitic and tumor antigens has been reported to trigger protective immunity. Objective: To evaluate the use of a DNA immunization strategy for protection against anthrax, a plasmid was constructed. Methods: The partialsequence of protective antigen of Bacillus anthracis, amino acids 175-764, as a potent immunogenic targe...
متن کامل