Part:BBa_K1974012

T7Promoter+RBS+Sf1a+linker+6X His-Tag

Introduction:

Figure 1.T7 promoter+RBS+Sf1a+linker+His-tag+terminator

      By ligating the IPTG induced promoter T7 (BBa_ I712074), strong ribosome binding site (BBa_B0034), sf1a, linker, and the 6xHistag (BBa_ K1223006), we can express Sf1a, the toxin by IPTG induction .
      This year we create a revolutionary system that integrates biological pesticides, an automatic detector, a sprinkler, and IoT. We made a database that contains most of the spider toxins and selected the target toxins by programming. U2-segestritoxin-Sf1a is coded for the venom of a spider, Segestria florentina. It is under the control of the strong T7 promoter. A 6xHistag is added for further protein purification.

Mechanism of Sf1a

      U2-segestritoxin-Sf1a has a structure called ICK(inhibitor cysteine knot). This kind of structure contains four disulfide bonds. With this structure, Sf1a can resist the high temperature, acid-base solution and the digest juice of insect gut. Sf1a can bind on insect voltage-gated sodium channel Site-1, making it paralyze and die eventually.

Features of Sf1a

1. Non-toxic: U2-segestritoxin-Sf1a is non-toxic to mammals and bees. Since the structure of the target ion channel is different, U2-segestritoxin-Sf1a does not harm mammals and bees. So it is safe to use it as a biological pesticide.

2. Biodegradable: The toxin is a peptide, so it must degrade over time. After degradation, the toxin will become nutrition in the soil.

3. Species-specific:According to reference, U2-segestritoxin-Sf1a has specificity to Lepidopteran (moths) and Dipteran (flies). So another insect such as bees will not be killed.

4. Eco-friendly: Compare with a chemical pesticide, U2-segestritoxin-Sf1a will not remain in soil and water so that it will not pollute the environment and won’t harm the ecosystem.

      Together, using Sf1a is totally an environmentally friendly way for solving harmful insect problems by using this ion channel inhibitor as a biological pesticide.

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]