DERB
Encyclopedia
Dual expression recombinase based (DERB) single vector system is a method of efficient cloning and subcloning of plasmid vectors for high throughput screening (HTS) and verification of protein-protein interactions inside living cells
. DERB was developed by Lu JP et al.
Vector
s are deliberately-constructed circular double-strand DNA loops capable of self-amplification and protein production. They are widely used in laboratories and the bio-medical and pharmaceutical industries to produce of DNA or protein in quantity. The DERB vector system consists of a series of vectors, each of which produces two or more proteins which are labeled or tagged for screening and verification of molecular interactions such as protein–protein interactions.
The following characteristics of the vectors ensure highly efficient cloning or subcloning of the protein of interest ORF
s into the DERB vectors for high-throughput screening (HTS) and verification of protein–protein interactions after single introducing of the vectors into eukaryotic or prokaryotic cells:
The system eliminates the requirements of multiple step restriction-purification-ligation subcloning and co-transfection, which are bottle-neck in automation in HTS protein–protein interactions.
The examination of the protein–protein interactions of interest with the DERB vector system can use standard laboratory equipment, e.g. plate reader, flow cytometer, fluorescent microscope and confocal microscope.
DERB is a new tool in high-throughput screening (HTS) and verification of protein–protein interactions and their modulators for both basic research and development such as screen new drugs or medicines for prevention and treatment of disease.
Cell (biology)
The cell is the basic structural and functional unit of all known living organisms. It is the smallest unit of life that is classified as a living thing, and is often called the building block of life. The Alberts text discusses how the "cellular building blocks" move to shape developing embryos....
. DERB was developed by Lu JP et al.
The DERB process
PlasmidPlasmid
In microbiology and genetics, a plasmid is a DNA molecule that is separate from, and can replicate independently of, the chromosomal DNA. They are double-stranded and, in many cases, circular...
Vector
Vector (molecular biology)
In molecular biology, a vector is a DNA molecule used as a vehicle to transfer foreign genetic material into another cell. The four major types of vectors are plasmids, viruses, cosmids, and artificial chromosomes...
s are deliberately-constructed circular double-strand DNA loops capable of self-amplification and protein production. They are widely used in laboratories and the bio-medical and pharmaceutical industries to produce of DNA or protein in quantity. The DERB vector system consists of a series of vectors, each of which produces two or more proteins which are labeled or tagged for screening and verification of molecular interactions such as protein–protein interactions.
The following characteristics of the vectors ensure highly efficient cloning or subcloning of the protein of interest ORF
Open reading frame
In molecular genetics, an open reading frame is a DNA sequence that does not contain a stop codon in a given reading frame.Normally, inserts which interrupt the reading frame of a subsequent region after the start codon cause frameshift mutation of the sequence and dislocate the sequences for stop...
s into the DERB vectors for high-throughput screening (HTS) and verification of protein–protein interactions after single introducing of the vectors into eukaryotic or prokaryotic cells:
- The sequences of two or more separate sets of promoters.
- Reporter tags for FRETFretA fret is a raised portion on the neck of a stringed instrument, that extends generally across the full width of the neck. On most modern western instruments, frets are metal strips inserted into the fingerboard...
, BiFC, PFCPFC- Science :*Perfluorinated compounds, a class of organofluorine compounds that have all hydrogen replaced with fluorine*Perfluorocarbons, fluorocarbon compounds derived from hydrocarbons...
etc. detection. - Recombinase recognizable sites for the insertion of ORFOpen reading frameIn molecular genetics, an open reading frame is a DNA sequence that does not contain a stop codon in a given reading frame.Normally, inserts which interrupt the reading frame of a subsequent region after the start codon cause frameshift mutation of the sequence and dislocate the sequences for stop...
of the proteins of interest. - Selection securities for efficient cloning.
The system eliminates the requirements of multiple step restriction-purification-ligation subcloning and co-transfection, which are bottle-neck in automation in HTS protein–protein interactions.
The examination of the protein–protein interactions of interest with the DERB vector system can use standard laboratory equipment, e.g. plate reader, flow cytometer, fluorescent microscope and confocal microscope.
DERB is a new tool in high-throughput screening (HTS) and verification of protein–protein interactions and their modulators for both basic research and development such as screen new drugs or medicines for prevention and treatment of disease.