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Recombinant proteins are a new combination of genes that forms DNA. Recombinant DNA technology allows for the production of wild type and modified human and mammalian proteins at bulk quantities. Recombinant proteins are made from cloned DNA sequences which usually encode an enzyme or protein with known function
Recombinant proteins are made through genetic engineering, also called gene splicing or recombinant DNA technology. By putting human, animal or plant genes into the genetic material of bacteria, mammalian or yeast cells, these microorganisms can be used as factories or producers to make proteins for medical, academic and research uses.
A vector is simply a tool for manipulating DNA and can be viewed as a "transport vehicle" for the production of proteins from specific DNA sequences cloned into them. Purification and expression of a protein can sometimes be quite complicated & time-consuming, therefore an additional tag is used in addition to the specific DNA sequence which will facilitate the purification & expresion of the recombinant protein.
Recombinant Proteins are proteins that their DNA that has been created artificially. DNA from 2 or more sources which is incorporated into a single recombinant molecule. The DNA is first treated with restriction endonuclease enzyme which the ends of the cut have an overhanging piece of single-stranded DNA. These are called "sticky ends" because they are able to base pair with any DNA molecule containing the complementary sticky end. DNA ligase covalently links the two strands into 1 recombinant DNA molecule.
Recombinant DNA molecule must be replicated many times to provide material for analysis & sequencing. Producing many identical copies of the same recombinant DNA molecule is called cloning. Cloning is done in vitro, by a process called the polymerase chain reaction (PCR). Cloning in vivo can be done in unicellular microbessuch as E. coli, unicellular eukaryotes like yeast and in mammalian cells grown in tissue culture.
Recombinant DNA must be taken up by the cell in a form in which it can be replicated and expressed. This is achieved by incorporating the DNA in a vector. A number of viruses (both bacterial and of mammalian cells) can serve as vectors.
Recombinant DNA is also sometimes referred to as chimera. When combining two or more different strands of DNA.There are 3 different methods by which Recombinant DNA is made. 1. Transformation, 2. Phage-Transfection 3.Yeast, Plant & Mammalian Transformation. When using the method of transformation one needs to select a piece of DNA to be inserted into a vector, cut a piece of DNA with a restriction enzyme and ligate the DNA insert into the vector with DNA Ligase. The insert contains a selectable marker which allows for identification of recombinant molecules. An antibiotic marker is used in order to cause death for a host cell which does not contain the vector when exposed to a certain antibiotic.
Trasnformation is the insertion of the vector into the host cell. The host cells are prepared to take up the foreign DNA. Selectable markers are used for antibiotic resistance, color changes, or any other characteristic which can distinguish transformed hosts from untransformed hosts. Yeast, Plant & Mammalian Transformation is done by micro-injecting the DNA into the nucleus of the cell being transformed. Phage-Transfection process, is equivalent to transformation except for the fact that phage lambda or MI3 is used instead of bacteria.
These phages produce plaques which contain recombinant proteins which can be easily distinguished from the non-recombinant proteins by various selection methods.
Significant amounts of recombinant protein are produced by the host only when expression genes are added. The Protein’s expression depends on the genes which surround the DNA of interest, this collection of genes act as signals which provide instructions for the transcription and translation of the DNA of interest by the cell. These signals include the promoter, ribosome binding site, and terminator.
The recombinant DNA is inserted into expression vectors which contain the promoter, ribosome binding site, and terminator.
In prokaryotic systems, the promoter, ribosome binding site, and terminator have to be from the same host since the bacteria is unlikely to understand the signals of human promoters and terminators. The designated gene must not contain human introns since the bacteria does not recognize it and this results in premature termination, and the recombinant protein may not be processed correctly, be folded correctly, or may even be degraded.
The peptide sequence can be added as an extension at the N-terminal. Researchers can select the specific purification system which they would like to use. The unique vectors available contain several features needed for the production of bulk quantities of the target protein. The peptide sequence is usually placed in the vector so that it is designed to be a point of attack for a specific protease. Thus, after the recombinant protein is expressed and extracted from bacteria, specific peptide extension can be used to purify the protein and subsequently removed from the target protein to generate a nearly natural sequence on the final product.
6 or more consistent Histidine residues act as a metal binding site for recombinant protein purification and expression. The hexa-His sequence is called a His-Tag sequence which can be placed on the N-terminal of a target protein by using vectors from various commercial molecular biology companies. The His-Tag contains a cleavage site for a specific protease. His-Tag recombinant proteins are purified by Metal Chelate Affinity Chromatography such as nickel ion columns that are used as the heavy metal ion and the His-Tag protein is eluted from the metal-chelate column with Histidine or imidazole. Then the purified His-Tag protein is treated with the specific protease to cleave off the His-Tag or not if the tag doesn’t affect the active site of the protein.
Proteins have metal binding sites which can be used for the purification of recombinant and natural proteins. This type of purification is rather simple when using a gel bead which is covalently modified so that it displays a chelator group for binding a heavy metal ion like Ni2+ or Zn2+. The chelating group on the gel bead contains a small amount of the ligands needed to hold the metal ion. So when the protein’s metal binding site finds the heavy metal, it will bind by providing the ligands from its metal binding site to attach to the metal ion displayed on the chelator location of the gel bead. This purification method is quite identical to affinity chromatography when purifying metal-binding class of proteins.
Recombinant Human Actin Gamma 1
Recombinant Human Actin-Like 6A
Recombinant Human Adipose Differentiation-Related Protein
Recombinant Mycobacterium Tuberculosis major secretory protein Antigen 85B
Recombinant Human Anterior Gradient Protein 2 Homolog
Recombinant Human Alpha-Internexin
Recombinant Human ALK-P80
Recombinant Human Angiostatin Kringles 1-3
Recombinant Human Annexin 7
Recombinant Human Annexin 9
Recombinant Human Assembly Protein Complex 2
Recombinant Human Assembly Protein Complex 3
Recombinant Human Amyloid P Component
Recombinant Human Autophagy Related 5
Recombinant Human APO-D
Recombinant Bovine Aprotinin
Recombinant Human Alpha-Synuclein
Recombinant Human B2 Microglobin
Recombinant Human b2 Glycoprotein-I
Recombinant Human BAD
Recombinant Human Bcl-2 (1-218)
Recombinant Human Bcl-2 Alpha
Recombinant Human Bcl-6
Recombinant Mouse Bax
Recombinant Human Beta-Trace
Recombinant Human Transforming Growth Factor Beta Induced protein
Recombinant Human BP-1
Recombinant Human Beta-Synuclein
Recombinant Human Cadherin-E
Recombinant Human Cadherin-N
Recombinant Rat Calbindin D-28K
Recombinant Rat Calbindin D-29K
Recombinant Rat Calbindin D-9K
Recombinant Human Calmodulin
Recombinant Murine Calretinin
Recombinant Human Caveolin-3
Recombinant Human CD10
Recombinant Human CD11b
Recombinant Human CD14
Recombinant Mouse CD14
Recombinant Human CD146
Recombinant Human CD147
Recombinant Human CD14 CHO derived
Recombinant Human CD1A
Recombinant Human CD2
Recombinant Human CD21
Recombinant Human CD23
Recombinant Human CD25
Recombinant Human CD29
Recombinant Human CD31
Recombinant Human CD34
Recombinant Human CD38
Recombinant Human CD43
Recombinant Human CD45
Recombinant Human CD52
Recombinant Human CD56
Recombinant Human CD68
Recombinant Human CD7
Recombinant Human CD72
Recombinant Human CD9
Recombinant Human CD95
Recombinant Human CD95, His Tag
Recombinant Human CD99
Recombinant Human Cell Division Cycle 6
Recombinant Human Carcinoembryonic Antigen
Recombinant Human CCAAT/enhancer binding protein(C/EBP) Alpha
Recombinant Human CCAAT/enhancer binding protein(C/EBP)
Gamma
Recombinant Human Centromere Protein-A
Recombinant Human Centromere Protein-B
Recombinant Human Chromagranin A
Recombinant Human Cbp/p300-Interacting TransActivator/MSP-1
Recombinant Human Cytokeratin 14
Recombinant Human Cytokeratin 18
Recombinant Human Cytokeratin 19
Recombinant Human Cytokeratin 20
Recombinant Human Cytokeratin 8
Recombinant Human Cofilin-1
Recombinant Human Like Collagen
Recombinant Human Collagen-1
Recombinant Human Collagen-3
Recombinant Human Collagen-VI
Recombinant Human Collapsin Response Mediator Protein-1
Recombinant Human C-Reactive Protein
Recombinant Human Cathepsin-D
Recombinant Human Cathepsin-L
Recombinant Rat Cystatin C
Recombinant Human D4-GDI
Recombinant Human Desmin
Recombinant Human DJ-1
Recombinant Human Down Syndrome Critical Region 1
Recombinant Human Endostatin
Recombinant Human Ephrin A1
Recombinant Human Ephrin A4
Recombinant Human Ephrin B1
Recombinant Human Ephrin B3
Recombinant Human Estrogen Receptor Alpha
Recombinant ESAT-6
Recombinant Human E-Selectin
Recombinant Human Ets-1
Recombinant Human Fatty Acid Binding Protein-1
Recombinant Human Fatty Acid Binding Protein-3
Recombinant Human Fatty Acid Binding Protein-4
Recombinant Human Fatty Acid Binding Protein-5
Recombinant Human Factor VIIa
Recombinant Human Factor VIII
Recombinant Human Factor VIII Related Antigen
Recombinant Human Fragile Histidine Triad
Recombinant Human Filaggrin
Recombinant Human FOS-B
Recombinant Human Fyn-p59
Recombinant Human GADD153
Recombinant Drosophila melanogaster GAGA-POZ
Recombinant Human Glomerular Basement Membrane
Recombinant Human Gross Cystic Disease Fluid Protein-15
Recombinant Human Gelatin
Recombinant Human Geminin
Recombinant Glutathione S-Transferase
Recombinant Human Gamma-Synuclein
Recombinant Human Haptoglobin
Recombinant Human Hypoxia-Inducible Factor-1 Alpha
Recombinant Hirudin
Recombinant Human High-Mobility Group Box 1
Recombinant Human Prostate Specific Gene-1
Recombinant Human Serum Albumin
Recombinant Human Serum Albumin mammalian derived
Recombinant Human Serum Albumin, Plant
Recombinant Human Intercellular Adhesion Molecule-1
Recombinant Human Intrinsic Factor
Recombinant Human Involucrin
Recombinant Human IPP-POZ
Recombinant Human IRF4 / MUM-1
Recombinant Human Keratin-8
Recombinant Human Killer Cell Immunoglobulin-Like Receptor, 2 Domains Long Cytoplasmic Tail 1
Recombinant Human Killer Cell Immunoglobulin-Like Receptor, 2 Domains Long Cytoplasmic Tail 3
Recombinant Human Killer Cell Immunoglobulin-Like Receptor, 2 Domains Short Cytoplasmic Tail 4
Recombinant Human Killer Cell Immunoglobulin-Like Receptor, 3 Domains Long Cytoplasmic Tail 1
Recombinant Human Ku P70/P80
Recombinant Human LA / SS-B
Recombinant Human Leukocyte-Associated Ig-Like Receptor 1
Recombinant Limulus Antilipopolysaccharide Factor
Recombinant Human Lactoferrin Holo
Recombinant Human Lactoferrin Apo
Recombinant Human Lipopolysaccarid Binding Protein
Recombinant Mouse Lipopolysaccarid Binding Protein
Recombinant Listeriolysin-O
Recombinant Listeriolysin-O PEST-free
Recombinant Human L-Selectin
Recombinant Human MAGE-1
Recombinant Human Mammaglobin
Recombinant Human MART-1/Melanocyte Differentiation-A
Recombinant Human Melanoma (gp100)
Recombinant Human MHC class I chain-related gene A
Recombinant Human MHC class I chain-related gene B
Recombinant Human Microphtalmia
Recombinant Human MutL Homolog-1
Recombinant Human MutS Homolog 2
Recombinant Human Myogenic Factor-4
Recombinant Human Myoblast Determination Protein 1
Recombinant Human Myoglobin
Recombinant Human Myoglobin Heme free
Recombinant Human Natural Cytotoxicity Receptor NKp46
Recombinant Human Non-Muscle Myosin-II Regulatory Light Chain
Recombinant Human New York Esophageal Squamous Cell Carcinoma
Recombinant Bacterial Outer Membrane Protein-A
Recombinant Human Osteocrin
Recombinant Human P21-WAF1
Recombinant Human P504/alpha Methylacyl CoA Racemase
Recombinant Human p53
Recombinant Human p53 Mutant
Recombinant Human p63
Recombinant Human Pancreatitis-Associated Protein-1
Recombinant Rat Parvalbumin
Recombinant Human Proliferating Cell Antigen
Recombinant Human Pancreas S Duodenum Homeobox-1
Recombinant Human Peripherin
Recombinant Human C-terminal hemopexin-like domain of MMP-2 (445-635 a.a.)
Recombinant Human p-Glycoprotein/MRP
Recombinant Human Ubiquitin Carboxyl-Terminal Hydrolase L1 Recombinant Human Ubiquitin Carboxyl-Terminal Hydrolase L1 GST tag
Recombinant Human PGR
Recombinant Human PM/Scl 100
Recombinant Human Procathepsin K
Recombinant Protein G
Recombinant Human P-Selectin
Recombinant Human PTEN
Recombinant Human RAB5A
Recombinant Human Ras-Related C3 Botulinum Toxin substrate 1
Recombinant Human Retinoblastoma Associated Protein
Recombinant Human Recoverin
Recombinant Human Regenerating Protein 1 alpha
Recombinant Human Regenerating Protein 1 beta
Recombinant Human Regenerating Protein IV
Recombinant Human Retinoid X Receptor Alpha
Recombinant Human RO-52/SS-A
Recombinant Human RO-60/SS-A
Recombinant Human Ribosomal Phosphoprotein P0
Recombinant Rat Receptor Associated Protein
Recombinant Human S100A1
Recombinant Human S100A10
Recombinant Human S100A11
Recombinant Human S100A4
Recombinant Human S100b
Recombinant Human S100P
Recombinant Human S100Z
Recombinant Human Sentrin-2
Recombinant Human Synaptosomal-associated protein 25kDa
Recombinant Synaptosomal-associated protein 25kDa, C.elegans
Recombinant Human Superoxide Dismutase
Recombinant Staphylococcal Protein-A
Recombinant Human Secreted Protein Acidic & Rich in Cysteine
Recombinant Spectrin-alpha
Recombinant Mouse Schmidt-Ruppin A-2 (SH3 Domain)
Recombinant Human Stefin A
Recombinant Streptavidin-NC
Recombinant Human Syntaxin-1A
Recombinant Human Small Ubiquitin-Related Modifier 1
Recombinant Human Small Ubiquitin-Related Modifier 2
Recombinant Human Syndecan-4
Recombinant Tobacco Etch Virus Protease
Recombinant Human Transcription Factor Binding to ighm Enhancer 3
Recombinant E.Coli Thioredoxin
Recombinant Human Thioredoxin
Recombinant Yeast Thioredoxin
Recombinant Human Thrombospondin
Recombinant Rat Tropic 1808
Recombinant Human Tumor Protein D52 L1
Recombinant Human TransGlutaminase-2
Recombinant Human fast skeletal Troponin 1
Recombinant Human Cardiac Troponin I-C Complex
Recombinant Single Chain Cardiac Troponin I-C
Recombinant Single Chain Cardiac Troponin I-C 2nd generation
Recombinant Human Cardiac Troponin I-T-C Complex
Recombinant Human Cardiac Troponin-C
Recombinant Human Cardiac Troponin-I
Recombinant Bovine Trypsin
Recombinant Human Thyroid Transcription Factor-1
Recombinant Human U1-snRNP 68kDa
Recombinant Human Ubiquitin
Recombinant Human Ubiquitin Gly76 to Ala76 mutation
Recombinant Human Ubiquitin Lys 48 to Arg 48 mutation
Recombinant Human Urokplakin-3
Recombinant Human Synaptobrevin-1
Recombinant Human Synaptobrevin-2
Recombinant Human Vascular cell adhesion molecule 1
Recombinant Human Von Hippel-Lindau Protein
Recombinant Human Vimentin
Recombinant Human Ventricular Myosin Light Chain LCII
Recombinant Human Yamaguchi Sarcoma Virus Oncogene Homolog 1
Recombinant Human Zinc-Alpha 2 Glycoprotein
Recombinant Human Zeta Chain Associated Protein kinase 70
Recombinant Human Zona Occludens 1 | 
