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Recombinant Proteins(重组蛋白)

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.

Products for  Recombinant Proteins

  1. Cat.No. 产品名称 Information
  2. GP23789 LAIR1 Human Leukocyte-Associated Ig-Like Receptor 1 Human Recombinant
  3. GP23788 L1CAM Human L1 Cell Adhesion Molecule Human Recombinant
  4. GP23787 LAGE3 Human L Antigen Family Member 3 Human Recombinant
  5. GP23786 LA/SS-B Human, Biotin LA / SS-B Human Recombinant, Biotinylated
  6. GP23785 LA/SS-B Human LA / SS-B Human Recombinant
  7. GP23784 KXD1 Human KxDL Motif Containing 1 Human Recombinant
  8. GP23783 Ku P70/P80 Human Ku P70/P80 重组人
  9. GP23782 KRT5 Human Cytokeratin 5 Human Recombinant
  10. GP23781 KRT8 Human, His Cytokeratin 8 Human Recombinant, His Tag
  11. GP23780 KRT8 Human, GST Cytokeratin 8 Human Recombinant, GST Tag
  12. GP23779 KRT8 Human

    Cytokeratin 8 Human Recombinant

  13. GP23778 KRT20 Human, His Cytokeratin 20 Human Recombinant, His Tag
  14. GP23777 KRT20 Human Cytokeratin 20 Human Recombinant
  15. GP23776 KRT19 Human, His Cytokeratin 19 Human Recombinant , His Tag
  16. GP23775 KRT19 Human

    Cytokeratin 19 Human Recombinant

  17. GP23774 KRT18 Human, His Cytokeratin 18 Human Recombinant , His Tag
  18. GP23773 KRT18 Human

    Cytokeratin 18 Human Recombinant

  19. GP23772 KRT17 Human Cytokeratin 17 Human Recombinant
  20. GP23771 KRT16 Human Cytokeratin 16 Human Recombinant
  21. GP23770 KRT14 Human, His Cytokeratin 14 Human Recombinant, His Tag
  22. GP23769 KRT14 Human Cytokeratin 14 Human Recombinant
  23. GP23768 KRAS 2A Human Kirsten Rat Sarcoma Viral Oncogene, Isoform 2A Human Recombinant
  24. GP23767 KRAS 2B Human Kirsten Rat Sarcoma Viral Oncogene, Isoform 2B Human Recombinant
  25. GP23766 KPNB1 Human Karyopherin Beta 1 Human Recombinant
  26. GP23765 KPNA2 Human Karyopherin Alpha 2 Human Recombinant
  27. GP23764 KLRK1 Human, Sf9 Killer Cell lectin-Like Receptor Subfamily K, Member 1 Human Recombinant, Sf9
  28. GP23763 KLRK1 Human Killer Cell lectin-Like Receptor Subfamily K, Member 1 Human Recombinant
  29. GP23762 KLRG1 Human Killer Cell Lectin-like Receptor Subfamily G, Member 1 Human Recombinant
  30. GP23761 KLRD1 Human Killer Cell Lectin Like Receptor D1 Human Recombinant
  31. GP23760 KLRC3 Human Killer Cell Lectin-Like Receptor Subfamily C, Member 3 Human Recombinant
  32. GP23759 KLRC2 Human Killer Cell Lectin-Like Receptor Subfamily C, Member 2 Human Recombinant
  33. GP23758 KLRC1 Human Killer Cell Lectin-Like Receptor Subfamily C, Member 1 Human Recombinant
  34. GP23757 KLRB1 Human, Sf9 Killer Cell Lectin-Like Receptor Subfamily B, Member 1 Human Recombinant, Sf9
  35. GP23756 KLRB1 Human Killer Cell Lectin-Like Receptor Subfamily B, Member 1 Human Recombinant
  36. GP23755 KLHDC8B Human Kelch Domain Containing 8B Human Recombinant
  37. GP23754 KLF12 Human Kruppel-Like Factor 12 Human Recombinant
  38. GP23753 KLF7 Human Kruppel-Like Factor 7 Human Recombinant
  39. GP23752 KLF6 Human Kruppel-Like Factor 6 Human Recombinant
  40. GP23751 KLF4 Human, His Kruppel-Like Factor 4 Human Recombinant, His Tag
  41. GP23750 KLF4 Human Kruppel-Like Factor 4 Human Recombinant
  42. GP23749 KLF3 Human Kruppel-Like Factor 3 Human Recombinant
  43. GP23748 KISS1 Human KISS-1 Metastasis-Suppressor Human Recombinant
  44. GP23747 KIR2DL5A Human Killer Cell Immunoglobulin-Like Receptor, 2 Domains Long Cytoplasmic Tail 5A Human Recombinant
  45. GP23746 KIR2DL4 Human Killer Cell Immunoglobulin-Like Receptor, 2 Domains Long Cytoplasmic Tail 4 Human Recombinant
  46. GP23745 KIR3DL2 Human Killer Cell Immunoglobulin-Like Receptor, 3 Domains Long Cytoplasmic Tail 2 Human Recombinant, Sf9
  47. GP23744 KIR3DL1 Human Killer Cell Immunoglobulin-Like Receptor, 3 Domains Long Cytoplasmic Tail 1 Human Recombinant
  48. GP23743 KIR2DS4 Human Killer Cell Immunoglobulin-Like Receptor, 2 Domains Short Cytoplasmic Tail, 4 Recombinant Human
  49. GP23742 KIR2DL3 Human Killer Cell Immunoglobulin-Like Receptor, 2 Domains Long Cytoplasmic Tail 3 Human Recombinant
  50. GP23741 KIR2DL1 Human Killer Cell Immunoglobulin-Like Receptor, 2 Domains Long Cytoplasmic Tail 1 Human Recombinant
  51. GP23740 KIN Human KIN Human Recombinant

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