What is the difference between pcDNA3 and pcDNA3 1?
pcDNA3 is no longer available from Thermo Fisher Scientific but has been directly replaced by pcDNA3. 1, which was derived from pcDNA3. The center of the multiple cloning site (MCS) within the original pcDNA3 vector contained homology to a hairpin mRNA structure and involved the Eag I, Not I, and both BstXI sequences.
What is pcDNA3 plasmid?
pcDNA3. Description: Mammalian expression vector with CMV promoter, BGH polyA signal, T7, SP6; amp resistance (neomycin for mammalian selection); restriction enzyme cloning. Synonyms: pcDNA-3.
What is pcDNA3?
Plasmid: pcDNA3 Notes: Mammalian expression vector with the CMV promoter and a neomycin-resistance marker.
What is pcDNA used for?
pcDNA3. 0 is a mammalian expression vector of 5.4 kb which are specially designed for high-level stable and transient expression in mammalian hosts. pcDNA is available with the multiple cloning sites in the forward (+) and reverse (–) orientations to facilitate good cloning.
What are pET vectors?
The pET vector system is a powerful and widely used system for expressing recombinant proteins in E. coli. The gene of interest is cloned into the pET vector under the control of the strong bacteriophage T7 transcription and translation regulatory system. Use of T7 RNA polymerase to direct expression of cloned genes.
What is a CMV promoter?
The CMV promoter is a commonly used promoter for the production of high level recombinant protein in mammalian cells17. However, the expression level of the transgene driven by CMV promoter decreases with extended culture times because of transcriptional silencing, which is associated with DNA methylation18, 19.
What is a multiple cloning site in a plasmid?
Multiple Cloning Site (MCS) Short segment of DNA which contains several restriction sites allowing for the easy insertion of DNA. In expression plasmids, the MCS is often downstream from a promoter. Insert. Gene, promoter or other DNA fragment cloned into the MCS for further study.
How does plasmid cloning work?
In a typical cloning experiment, a target gene is inserted into a circular piece of DNA called a plasmid. The plasmid is introduced into bacteria via a process called transformation, and bacteria carrying the plasmid are selected using antibiotics.
What is pLysS?
pLysS is a plasmid that contains the T7 lysozyme gene (LysS). The T7 lysozyme binds to T7 RNA polymerase causing inhibition until induction by the addition of IPTG. When IPTG is added, the amount of T7 RNA polymerase increases and over- comes the inhibition by LysS.
What are the features of the pcdna3.1 vector family?
The pcDNA™3.1 Expression Vector Family Three untagged versions of pcDNA™3.1 (available separately), each with a different selectable marker (Geneticin®, Zeocin™, or Hygromycin), are for use alone or in co-transfections. All three vectors offer the following features: • Cytomegalovirus (CMV) enhancer-promoter for high-level expression
Are there any untagged versions of pcdna3.1?
Three untagged versions of pcDNA™3.1 (available separately), each with a different selectable marker (Geneticin®, Zeocin™, or Hygromycin), are for use alone or in co-transfections. All three vectors offer the following features: For Research Use Only. Not for use in diagnostic procedures.
How is pcdna3.1 used to detect recombinant proteins?
The pcDNA3.1 vector delivers high-level expression of transient or stable GFP fusion proteins in a wide range of mammalian cells Expression is easily detected in living cells using fluorescence. In addition, recombinant proteins expressed from these vectors can be detected on western blots using GFP Antiserum.
Which is the empty backbone of pcdna3 plasmid?
(Empty Backbone) This pcDNA3 plasmid contains a new poly-linker and single C-terminal flag-tag sequence for expression in mammalian cells. Sanjabi et al Genes Dev. 2005 Sep 15. 19 (18):2138-51.