Part:BBa_K4632003

Cowpea trypsin inhibitor(CPTI)

Description

1. How does it work? CPTI can inhibite the serine protease by interacting with the active site of serine protease. Simultaneously, CPTI inhibits insect feeding by stimulating feedback signals from insect (Xuhong-lin et al.,2008)

2. Eco-friendly and Safe CPTI, a member of the Bowman-Birk protein family. Anti-insect spectrum tests show that CPTI inhibits almost all tested major agricultural pests. (Xuhong-lin et al.,2008) And, most importantly. it is Eco-friendly and Safe. (see more detail on https://2023.igem.wiki/scau-china/safety）

3. What we have done? (SCAU-China 2023) Differing from the original publication (Wei Gao et al., 2013), we did not fuse the CPTI gene with a GST purification tag for expression. Our aim is to utilize CPTI expression directly for the eradication of red fire ants, without the subsequent GST tag removal step. Furthermore, if CPTI without the GST tag can be successfully expressed, its protein molecular weight would be only 10.4 kDa.

After conducting Tricine-PAGE and Western Blot experiments, the results indicate that the CPTI gene without the GST tag does not express as expected, as described in the literature. In the next step, we have constructed the CPTI gene with the GST tag, and protein induction expression is currently underway.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Construction and Characterization

1. Verifying the Expression and Secretion Proficiency of CPTI