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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. GP26368 Tri a 14.0101 Recombinant Non-Specific Lipid-Transfer Protein Tri a 14 produced in SF9 is a glycosylated, polypeptide chain having a calculated molecular mass of 13kDa
  3. GP26367 Pen a 1.0101 Recombinant Tropomyosin Pen a 1
  4. GP26366 MALD3 Recombinant Non-Specific Lipid-Transfer Protein Mal d 3 produced in SF9 is a glycosylated, polypeptide chain having a calculated molecular mass of 12kDa
  5. GP26365 Cor a 14.0101 Recombinant 2S albumin produced in SF9 is a glycosylated, polypeptide chain having a calculated molecular mass of 14kDa
  6. GP26364 Cor a 9 The native protein Corylus Avellana Cor a 9 is purified from hazelnut by protein chemical methods
  7. GP26363 Cor a 8.0101 Recombinant Non-specific Lipid-Transfer Protein Cor a 8 produced in SF9 is a glycosylated, polypeptide chain having a calculated molecular mass of 11kDa
  8. GP26362 HSP27 Mouse HSP27 Mouse Recombinant produced in E
  9. GP26357 VAMP2 Human, (1-94) VAMP2 Human Recombinant produced in e
  10. GP26355 ULBP3 Human, Sf9 ULBP3 Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 430 amino acids (30-217a
  11. GP26354 TXN Mouse TXN Mouse Recombinant produced in E
  12. GP26353 Troponin C-I-T Complex Human Cardiac Troponin C-I-T Complex Protein produced in Human heart tissue having a molecular mass of approximately 75kDa
  13. GP26349 TFRC Native Human Transferrin Receptor Protein produced in human serum tissue having a molecular mass of 85kDa
  14. GP26347 RAGE Human, Sf9 RAGE Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 561 amino acids (24-342 a
  15. GP26346 SPON1 Human SPON1 Human Recombinant produced in HEK293 cells is a single, glycosylated, polypeptide chain (29-807 a
  16. GP26341 SOD Human, 15N Recombinant Human Superoxide Dismutase, 15N produced in E
  17. GP26340 SNCA E46K, Human SNCA E46K Human Recombinant produced in E
  18. GP26339 SNCA 1-95, Human SNCA Human Recombinant produced in E
  19. GP26337 SIRPA Human, HEK SIRPA Human Recombinant produced in HEK293 cells is a single, glycosylated polypeptide chain (27-373a
  20. GP26336 SIGLEC10 Human SIGLEC10 Human produced in Sf9 Insect cells is a single, glycosylated polypeptide chain containing 678 amino acids (17-455 a
  21. GP26334 SFRP2 Mouse SFRP2 Mouse Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 280 amino acids (25-295 a
  22. GP26333 SERPING1 Human, Native Human Serpin Peptidase Inhibitor, Clade G Member 1 produced in Human plasma having a molecular mass of 110 kDa
  23. GP26332 SERPINA7 Human Human Serpin Peptidase Inhibitor, Clade A Member 7 Protein produced in Human plasma having a molecular mass of approximately 55kD
  24. GP26331 SERPINA6 Human SERPINA6 Human Recombinant is a single, glycosylated polypeptide chain containing 389 amino acids (23-405a
  25. GP26329 SELE Human, Sf9 SELE Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 541 amino acids (22-556 a
  26. GP26328 RAET1L Human RAET1L Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 202 amino acids (26-218 a
  27. GP26326 Prealbumin Human Human Transthyretin dimer protein produced in Human plasma having a molecular mass of 30kD
  28. GP26321 PDCD1 Human, Active PDCD1 Human Recombinant produced in HEK293 Cells is a single, glycosylated polypeptide chain containing 383 amino acids (21-170 a
  29. GP26320 ORM1 Human, HEK ORM1 Human Recombinant produced in HEK is a polypeptide chain containing 189 amino acids (19-201) and having a molecular mass of 22
  30. GP26319 NME4 Human, Active NME4 Human Recombinant produced in E
  31. GP26318 NME3 Human, Active NME3 Human Recombinant produced in E
  32. GP26317 NME2 Human, Active NME2 Human Recombinant produced in E
  33. GP26316 NME1 Human, Active NME1 Human Recombinant produced in E
  34. GP26315 NECTIN1 Human NECTIN1 Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 334amino acids (31-355a
  35. GP26313 NCAM1 Human NCAM1 Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 593 amino acids (20-603 a
  36. GP26309 MORC3 Human Recombinant Human MORC Family CW-Type Zinc Finger 3 produced in SF9 is a glycosylated, polypeptide chain having a calculated molecular mass of 122kDa
  37. GP26302 LILRB2 Human LILRB2 Human Recombinant produced in HEK293 Cells is a single, glycosylated polypeptide chain containing 444 amino acids (24-461 a
  38. GP26301 LILRB1 Human LILRB1 Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 446 amino acids (24-461 a
  39. GP26300 KLRF1 Human KLRF1 Human Recombinant produced in HEK293 Cells is a single, glycosylated polypeptide chain containing 414 amino acids (60-231a
  40. GP26299 IgM Human Human Immunoglobulin-M produced in human plasma having a molecular mass of 950kDa
  41. GP26297 IgG1 Fc Human IgG1 Fc Human Recombinant produced in HEK is a single polypeptide chain containing 231 amino acids (100-330) and having a molecular mass of 25
  42. GP26296 IMPAD1 Human, Active IMPAD1 Human Recombinant produced in E
  43. GP26295 ICAM5 Human ICAM5 Human Recombinant produced in HEK293 is a single, glycosylated polypeptide chain containing 1052 amino acids (23-835 a
  44. GP26294 ICAM3 Human ICAM3 Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 698 amino acids (30-485 a
  45. GP26293 ICAM1 Mouse ICAM1 Mouse Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 466 amino acids (28-485 a
  46. GP26292 Gliadin Native The native Gliadin Triticum Aestivum Grain is purified from wheat by protein chemical methods
  47. GP26286 EPHB1 Human EPHB1 Human Recombinant produced in HEK293 Cells is a single, glycosylated polypeptide chain containing 529 amino acids (18-540 a
  48. GP26283 EIF3I Human, Sf9 EIF3I Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 331 amino acids (1-325 a
  49. GP26282 EFNA3 Human, Sf9 EFNA3 Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 434 amino acids (23-214aa) and having a molecular mass of 48
  50. GP26280 DKK1 Mouse DKK1 Mouse Recombinant produced in HEK293 cells is a single, glycosylated polypeptide chain (32-272a
  51. GP26278 CTHRC1 Human, Sf9 CTHRC1 produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 222 amino acids (31-243a

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