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Recombinant protein is a protein coded by the manipulated gene - Recombinant Gene and generated by the specific recombinant expression system (see protein expression systems). The recombinant gene is a new genetic combination that one or more DNA segments or genes have been inserted from a different molecule or from from other species. Compared with natural protein, recombinant protein can be produced in large amounts with relative ease.
Recombinant protein plays an important role in studying biological processes such as Cytokines and Growth Factors, enzymes and kinases researches and complement system functions. Moreover, recombinant proteins are known as highly potent medicines that are safe from off-target side effects, take a shorter time to develop than small molecules (see recombinant protein drugs).
To make recombinant proteins, the gene is isolated and cloned into an expression vector. Generating a recombinant protein requires the protein expression system, protein purification system and protein identification systems.
Basic steps to get recombinant Protein:
1. Amplification of gene of interest.
2. Insert into cloning vector.
3. Sub cloning into expression vector.
4. Transformation into protein expressing host (bacteria (E coli), yeast, mammalian cells or baculovirus-insect cell system).
5. Test for identification of recombinant protein.( Western blot or Fluoroscence)
6. Large scale production. (Large scale fermentor)
7. Isolation and purification.
Because the chemical and physical properties of proteins vary, they can be isolated from one another for subsequent analysis by several commonly used methods:
Centrifugation – A protein's mass and shape give it a specific rate of sedimentation under high g-forces, allowing separation
Gel electrophoresis – Polypeptide chains of different molecular weights migrate through a gel matrix (such as polyacrylamide) at different rates when an electric field is applied
Liquid chromatography – Proteins are separated based on their movement through a column containing spherical beads. Liquid chromatography allows separation by mass (gel filtration column), charge (ion-exchange column) and ligand-binding behavior (affinity column)
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Recombinant protein is a protein coded by the manipulated gene - Recombinant Gene and generated by the specific recombinant expression system (see protein expression systems). The recombinant gene is a new genetic combination that one or more DNA segments or genes have been inserted from a different molecule or from from other species. Compared with natural protein, recombinant protein can be produced in large amounts with relative ease.
Recombinant protein plays an important role in studying biological processes such as Cytokines and Growth Factors, enzymes and kinases researches and complement system functions. Moreover, recombinant proteins are known as highly potent medicines that are safe from off-target side effects, take a shorter time to develop than small molecules (see recombinant protein drugs).
To make recombinant proteins, the gene is isolated and cloned into an expression vector. Generating a recombinant protein requires the protein expression system, protein purification system and protein identification systems.
Basic steps to get recombinant Protein:
1. Amplification of gene of interest.
2. Insert into cloning vector.
3. Sub cloning into expression vector.
4. Transformation into protein expressing host (bacteria (E coli), yeast, mammalian cells or baculovirus-insect cell system).
5. Test for identification of recombinant protein.( Western blot or Fluoroscence)
6. Large scale production. (Large scale fermentor)
7. Isolation and purification.
Because the chemical and physical properties of proteins vary, they can be isolated from one another for subsequent analysis by several commonly used methods:
Centrifugation – A protein's mass and shape give it a specific rate of sedimentation under high g-forces, allowing separation
Gel electrophoresis – Polypeptide chains of different molecular weights migrate through a gel matrix (such as polyacrylamide) at different rates when an electric field is applied
Liquid chromatography – Proteins are separated based on their movement through a column containing spherical beads. Liquid chromatography allows separation by mass (gel filtration column), charge (ion-exchange column) and ligand-binding behavior (affinity column)