What is Dyk tag?
What is Dyk tag?
l DYK tag: A short peptide composed of 8 amino acids (DYKDDDDK), which can be fused to the N-terminus or C-terminus of the target protein and used for the immunoadsorption purification of recombinant protein.
What is pcDNA used for?
The control plasmid, pcDNA™3.1/CAT, is included for use as a positive control for transfection and expression in the cell line of choice.
What is pCMV plasmid?
The pCMV-Script vector can be rescued as single-stranded (ss) DNA. The plasmid contains a 454-nucleotide filamentous f1 phage intergenic region (M13-related) that includes the 307 bp origin of replication. The orientation of the f1 origin in pCMV-Script allows the rescue of antisense ssDNA by a helper phage.
What is pcDNA?
pcDNA (plural pcDNAs) (genetics) plasmid cloning DNA.
How do you purify the flag tagged protein?
How to Purify FLAG-tagged Proteins? FLAG-tagged recombinant protein can be affinity purified directly from a cell culture lysate or supernatant. The FLAG-tagged protein binds to the FLAG-tag specific monoclonal antibody conjugated on an agarose gel.
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 pCMV vector?
The pCMV-Script vector is derived from a high-copy-number pUC-based plasmid and is designed to allow protein expression in mammalian systems. Mammalian expression is driven by the human cytomegalovirus (CMV) immediate early promoter to promote constitutive expression of cloned inserts in a wide variety of cell lines.
What is a lentiviral plasmid?
When lentivirus is used for research, it is the lentiviral genome that encodes genetic material that the researcher wants delivered to specific target cells. This genome is encoded by plasmids called “transfer plasmids,” which can be modified to encode a wide range of gene products.
What is a mammalian expression vector?
Mammalian expression vectors are used to introduce a specific fragment of DNA into mammalian systems for RNA or protein expression. These systems include human and mouse cell lines, and even in vivo in live animals.
How do you remove flag peptides?
The tag can be removed by the action of proteases such as enterokinase, thrombin and factor-Xa. G- Biosciences recommends the use of highly effective recombinant enterokinase that can be used to remove the FLAG tag.