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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. GC66026 SIRT2-IN-9 SIRT2-IN-9 (compound 12) 是 SRIT2 的选择性抑制剂,IC50 值为 1.3 μM。SIRT2-IN-9 抑制 MCF-7 乳腺癌细胞的增殖活性。SIRT2-IN-9 可用于癌症的研究。
  3. GC65935 CAM833 CAM833 是一种有效的抑制 BRCA2 和 RAD51 之间相互作用的正位抑制剂,对 ChimRAD51 蛋白的 Kd 为 366 nM。 CAM833 也抑制 RAD51 的寡聚。
  4. GC65909 Bexotegrast Bexotegrast 是 αΝβ6 整合素的有效抑制剂。Bexotegrast 可用于研究特发性肺纤维化 (IPF) 和非特异性间质性肺炎 (NSIP) 等纤维化 (信息摘自专利 WO2020210404A1,compound 5)。
  5. GC52173 N-desmethyl-Doxylamine (succinate) A metabolite of doxylamine
  6. GC65597 Abciximab Abciximab (C7E3) 是一种小鼠/人嵌合单克隆抗体,是一种糖蛋白 IIb/IIIa(glycoprotein IIb/IIIa)抑制剂。 Abciximab 通过与糖蛋白 IIb/IIIa、vitronectin 和 Mac-1 受体结合抑制血小板聚集和白细胞粘附。
  7. GC65581 MIND4-19 MIND4-19 是一种有效的 SIRT2 抑制剂,IC50 为 7.0 μM。MIND4-19 可用于亨廷顿氏舞蹈症 (Huntington's disease) 的研究。
  8. GC65455 c(phg-isoDGR-(NMe)k) c(phg-isoDGR-(NMe)k) 是有效选择性的 α5β1 整合素 配体,IC50 为 2.9 nM。
  9. GC65383 RAD51-IN-2 RAD51-IN-2 (化合物 example 67A) 是一种 RAD51 抑制剂,详细信息请参考专利文献 WO2019/051465A1。
  10. GC65322 FABPs ligand 6 FABPs ligand 6 (MF6) 是一种 FABP5 和 FABP7 抑制剂,KD 值分别为 874 nM 和 20 nM。FABPs ligand 6 可用于多发性硬化症的研究。
  11. GC65030 Sirtuin modulator 2 Sirtuin modulator 2 (Compound 132) 是一种 sirtuin 调节剂,ED50 不超过 50 μM。
  12. GC65019 SRT 1460 SRT 1460 是一种有效的 Sirtuin-1 (SIRT1) 激动剂,其 EC1.5 值为 2.9 μM。与对 SIRT2 和 SIRT3 (EC1.5>300 μM) 相比,SRT 1460 对 SIRT1 有很好的选择性,并且比白藜芦醇和相近 sirtuin 同系物更为有效。
  13. GC64946 SRT 2183 SRT 2183 是一种选择性 Sirtuin-1 (SIRT1) 激活剂,其 EC1.5 值为 0.36 μM。SRT 2183 诱导生长停滞和凋亡,同时伴随 STAT3 和 NF-κB 去乙酰化,并降低 c-Myc 蛋白水平。
  14. GC64932 αvβ5 integrin-IN-1 αvβ5 integrin-IN-1 是第一个有效和选择性 αvβ5 整合素抑制剂(pIC50 = 8.2)。
  15. GC64575 Et-29 Et-29 是一种有效的,选择性的 SIRT5 抑制剂 (Ki=40 nM)。
  16. GC64571 Sibrafiban Sibrafiban (RO 48-3657) 是一种 Ro 44-3888 的具有口服活性的,非肽,双重前药,是一种选择性的糖蛋白 IIb/IIIa 受体 (glycoprotein IIb/IIIa receptor) 拮抗剂。Sibrafiban 可抑制血小板聚集。
  17. GC64527 ADTL-SA1215 ADTL-SA1215 是一种调节三阴性乳腺癌自噬的特异性 SIRT3 小分子激活剂。
  18. GC52105 Montelukast Montelukast (MK0476 free base) 是半胱氨酰白三烯受体 1 (CysLT1) 的有效、选择性和口服活性拮抗剂。
  19. GC52088 Methylchloroisothiazolinone (hydrochloride) A biocide
  20. GC52081 Chamazulene Chamazulene 是一种天然化合物,是一种抗氧化型的白三烯 B4 形成抑制剂。
  21. GC52074 Loratadine N-oxide Loratadine n-oxide 是氯雷他定的代谢物。
  22. GC52045 N-desmethyl Azelastine N-去甲基氮卓斯汀是氮卓斯汀的主要活性代谢物,经细胞色素P450酶系统氧化代谢,蛋白结合率为97%,消除半衰期为54小时。
  23. GC52043 Hydrocortisone Aceponate Hydrocortisone aceponate (Hydrocortisone 17-propionate 21-acetate) 是一种有效的外用糖皮质激素。
  24. GC52016 Cetirizine N-oxide An oxidative degradation product of cetirizine
  25. GC49884 Trigonelline-d3 (chloride) An internal standard for the quantification of trigonelline
  26. GC49755 Meclizine N-oxide A metabolite of meclizine
  27. GC49753 LCS3 LCS3 是一种可逆且非竞争性的谷胱甘肽二硫化物还原酶 (GSR) 和硫氧还蛋白还原酶 1 (TXNRD1) 抑制剂(IC50=3.3 μM 和 3.8 μM 分别)。 LCS3 显示出抗肿瘤活性,并诱导细胞凋亡。 LCS3 可用于肺腺癌 (LUAD) 研究。
  28. GC64255 CHIC35 CHIC35 是 EX-527 的结构类似物,是 SIRT1 (IC50=0.124 µM) 的有效选择性抑制剂。CHIC35 对 SIRT1 的选择性远大于对 SIRT2 (IC50=2.8 µM) 和 SIRT3 (IC50>100 µM)。CHIC35 具有抗炎作用,可用于 CHARGE 综合征的研究。
  29. GC64212 Valategrast hydrochloride Valategrast hydrochloride (R-411) 是一种有效的整联蛋白 α4β1 (VLA-4) 和 α4β7 双重拮抗剂。Valategrast hydrochloride 可用于慢性阻塞性肺疾病 (COPD) 和哮喘的研究。
  30. GC49724 BIO-1211 (trifluoroacetate salt) A peptide inhibitor of α4β1 integrin
  31. GC49676 6β-hydroxy Budesonide A metabolite of budesonide
  32. GC49674 Schizandrin 五味子素(Schizandrin),一种二苯并环辛二烯木脂素,从五味子的果实中分离得到。
  33. GC49652 Chlorpheniramine-d6 (maleate) An internal standard for the quantification of chlorpheniramine
  34. GC63490 Valategrast Valategrast (R-411 free base) 是一种有效的口服活性整联蛋白 α4β1 (VLA-4) 和 α4β7 双重拮抗剂。Valategrast 可用于慢性阻塞性肺疾病 (COPD) 和哮喘的研究。
  35. GC63165 RAD51-IN-1 RAD51-IN-1 是 B02 的衍生物,是 RAD51 的有效抑制剂。 RAD51-IN-1 可用于癌症研究。
  36. GC62886 Cannabisin F Cannabisin F 是一种 SIRT1 调制器。Cannabisin F 作为大麻籽木素酰胺,可用于抗炎、抗氧化研究。Cannabisin F 作为SIRT1/NF-κB 和 Nrf2 的调节因子,可能是一种潜在的神经退行性疾病的调节剂。
  37. GC62872 BMS-587101 BMS-587101 是一种有效的具有口服活性的 leukocyte function associated antigen-1 (LFA-1) 拮抗剂。BMS-587101 具有抗炎作用,可用于类风湿关节炎的研究。
  38. GC49433 Capsiate Capsiate 是一种从无刺激性 CH-19 甜红辣椒品种中提取的辣椒素类似物,是一种口服活性的 TRPV1 激动剂 。
  39. GC49420 Gallic Acid-d2 An internal standard for the quantification of gallic acid
  40. GC49419 Aniline-d5 An internal standard for the quantification of aniline
  41. GC49343 Isoproterenol-d7 (hydrochloride) An internal standard for the quantification of isoproterenol
  42. GC49309 A-943931 (hydrochloride hydrate) A histamine H4 receptor antagonist
  43. GC49294 1-(4-Chlorobenzhydryl)piperazine An inactive metabolite of meclizine and chlorcyclizine
  44. GC49197 Carbinoxamine-d6 (maleate) An internal standard for the quantification of carbinoxamine
  45. GC49138 Naphazoline-d4 (hydrochloride) An internal standard for the quantification of naphazoline
  46. GC49093 Safflower Red A red pigment with diverse biological activities
  47. GC49089 FR900359 A cyclic depsipeptide and an inhibitor of Gαq, Gα11, and Gα14<•sub>
  48. GC49082 Cicloprofen An NSAID
  49. GC49072 Tribenoside Tribenoside 是一种血管保护剂,可用于痔疮的研究。
  50. GC49063 Emedastine-13C-d3 (fumarate) An internal standard for the quantification of emedastine
  51. GC49062 Tiopronin-d3 硫普罗宁-d3 是氘标记的硫普罗宁。

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