Part:BBa_K4003006

pET-24a-PPM1A

Usage and Biology

Studies have shown that an Alzheimer’s patient is born every three seconds. As of 2019, more than 52 million people worldwide are living with Alzheimer’s disease. The patients in China achieved a quarter of the total. In the same year, an expert in Alzheimer’s research in the United States said,” They're maybe more people who go undiagnosed. Nowadays millions of people worldwide are suffered from the pain of Neurodegenerative diseases (NIH) and currently, no one in the world can completely cure these kinds of diseases, all we can do is slow down the process.

So our product is targeting those patients who suffer from the disease related to the microglia Inflammation, such as Alzheimer’s.

1. Purification principle of recombinant human PPM1A protein:

Fig1. ÄKTA™ pure protein purifier..

Six histidines. Histidine can specifically bind to nickel ions, and the recombinant protein with histidine tag can be adsorbed by the nickel column to achieve the purpose of purification. In addition, the eluted PPM1A protein is further separated and purified by ÄKTA™ pure protein purifier with Superdex 200 molecular sieve chromatography column. Finally, replace the storage buffer (50 mM Tris-HCl, pH 7.0, 10% glycerol, 1 mM DTT) through an ultrafiltration tube, concentrate the protein, and quickly freeze and store in liquid nitrogen.

Fig2. PPM1A enzyme activity screening principle..

pNPP (p-nitrophenyl phosphate disodium salt) is used as a substrate. When the compound and PPM1A protein are added, the enzyme activity reaction will be initiated. The absorbance at 410 nm is measured with a microplate reader. By calculating the reaction rate, the agonistic efficiency of the compound was evaluated.

Fig3. Inflammation of microglia..

Microglia will produce inflammatory factors such as IL-6 and TNFα under the stimulation of LPS.

BBa_K4003002

Usage and Biology

The promoter can react specifically to T7 RNA polymerase and is a sequence that starts gene transcription of T7 phage. It is recognized for binding and initiation of the transcription. Placed before a gene, it promotes its transcription.

BBa_K4003004

Usage and Biology

It is an polyhistidine tag, which is used in the purification of recombinant proteins

BBa_K4003000

Properties: Gene technology for protecting patented bacterial strains or cell

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 5075
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 4964
  Illegal BglII site found at 6045
  Illegal BamHI site found at 5108
  Illegal XhoI site found at 5148
  Illegal XhoI site found at 6516
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 137
  Illegal NgoMIV site found at 3184
  Illegal NgoMIV site found at 3344
  Illegal NgoMIV site found at 4932
  
• 1000
  COMPATIBLE WITH RFC[1000]