Part:BBa_K1298004

Added by LZU-HS-China-A

For the measurement of binding capacity

we took 2 μmol/L of chitinase in 50 mmol/L phosphate buffer (pH=8) and mixed thoroughly at 4°C (assuming no degradation), reacted for 1 h on a rotary mixer, 10,000 r/min for 5 min and collected the supernatant, which was the unbound protein, by Protein concentration was measured by the BCA method.

Figure 1 Binding rates

For the measurement of chitinase activity

chitinase is able to hydrolyse chitin to produce N-acetylglucosamine, which further reacts with 35-dinitrosalicylic acid to produce a brownish-red compound with a characteristic absorption peak at 540 nm, and the activity of chitinase can be characterised by the change in absorbance value.

Figure 2 Chitinase activity

PgeChia 1-1 (insert) using J04500

This part contains an RBS (BBa_B0034), a promoter (lacI inducible; BBa_R0010), and a coding region (PgeChia 1-1; BBa_K1298001). It is in a pSB1C3 backbone. With this, the chitinase protein is created. Chitinase breaks down chitin, a substance found in the exoskeleton of arthropods (spiders, insects, etc.) and in the cell walls of fungi. It should be noted that there is no secretory tag, meaning that the chitinase protein is not being secreted outside of the cell, but staying inside. The part should be used to have the chitinase protein break down chitin.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 296
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 822
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 682
  Illegal NgoMIV site found at 851
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 996