What is puro gene?
pSELECT-puro plasmids offer all the features necessary to express a gene of interest at high levels in a large number of cell types. pSELECT-puro plasmids contain two transcription units, the first drives the expression of the gene of interest and the second drives the expression of a puromycin selectable marker.
What is dual expression plasmid?
Dual-expression vectors for expression of proteins encoded by these genes in mammalian and bacterial cells can be used for this characterization. Typically, eukaryotic genes are expressed in mammalian cells to characterize biological functions and in bacterial cells to facilitate isolation of the protein.
How does puromycin resistance gene work?
Puromycin is a naturally occurring aminonucleoside antibiotic that inhibits protein synthesis by ribosome-catalyzed incorporation into the C-terminus of elongating nascent chains, blocking further extension and resulting in premature termination of translation.
Does T7 promoter work in mammalian cells?
The phage T7 promoter/polymerase system is highly specific in bacteria in contrast to that observed in mammalian cells. A number of cell lines exhibit a considerable level of expression from the T7 promoter, even in the absence of T7-RNA polymerase.
What is the purpose of puromycin?
Puromycin is an antibiotic that prevents bacterial protein translation. It is utilized as a selective agent in laboratory cell cultures. Puromycin is toxic to both prokaryotic and eukaryotic cells, resulting in significant cell death at appropriate doses.
Why is T7 promoter used?
The T7 promoter is commonly used to regulate gene expression of recombinant proteins, which can be subsequently used for a variety of downstream research applications2.
How does puromycin selection work?
This compound works by interrupting protein synthesis in both prokaryotic and eukaryotic cells. This causes cells to die, unless they have a gene for resistance, allowing them to survive exposure to the antibiotic.
Is puromycin stable in media?
Puromycin is stable at 4C but if you add puromycin into your cell culture media and do multiple “warmings” (i.e. constantly pre-warming your media), then the puromycin in the media will degrade.
What are lentiviruses used for?
Use as a vector Some experimental applications of lentiviral vectors have been done in gene therapy in order to cure diseases like Diabetes mellitus, Murine haemophilia A, prostate cancer, chronic granulomatous disease, and vascular diseases.