Part:BBa_K2599016

T7 Promoter+RBS+Sf1a+GS linker+snowdrop-lectin+linker+6X His-Tag

NCTU_Formosa 2018 designed this sequence to improve the part from NCTU_Formosa 2016.

We changed the general linker to a GS linker to optimize the function of Sf1a.

Previous Part:(BBa_K1974022)

The existing part from NCTU_Formosa 2016 contains the IPTG induced PT7 (BBa_I712074), strong ribosome binding site (BBa_B0034), Sf1a, AAA linker, snowdrop lectin (BBa_K1974020) and the 6X His-Tag (BBa_K1223006).

Figure 1. Previous part

Improvement part

In many application, the fusion proteins consisting of multiple protein domains is a popular and highly successful approach to engineering new protein functions. The linker that connects the fusion protein often plays an important role in fusion proteins. Therefore, we decided to change the linker between Sf1a and lectin to try whether the function of this protein become stronger. The GS linker we used in this improvement part was also provided by NCTU_Formosa 2016. It contained 18 amino acid sequence, which is a Gly-Gly-Ser repeated linker. We utilize this linker to optimize the fuction of Sf1a and lectin.

Figure 1. Improvement part

Introduction

Figure 1. Comparison of plant protecting effects

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]

Reference

1. Elaine Fitches, Martin G. Edwards, Christopher Mee, Eugene Grishin, Angharad M. R. Gatehouse, John P. Edwards, John A. Gatehouse “Fusion proteins containing insect-specific toxins as pest control agents: snowdrop lectin delivers fused insecticidal spider venom toxin to insect haemolymph following oral ingestion,” Journal of Insect Physiology, 2004, 50, pp.61-71
2. Elaine C. Fitches, Prashant Pyati, Glenn F. King, John A. Gatehouse, “ Fusion to Snowdrop Lectin Magnifies the Oral Activity of Insecticidal Omega-Hexatoxin-Hv1a Peptide by Enabling Its Delivery to the Central Nervous System,”
3. Monique J. Windley, Volker Herzig, Slawomir A. Dziemborowicz, Margaret C. Hardy, Glenn F. King and Graham M. Nicholson, “Spider-Venom Peptide as Bioinsecticide,” Toxins Review, 2012, 4, pp. 191-227.
4. A. Lipkin, S. Kozlov, E. Nosyreva, A. Blake, J.D. Windass, E. Grishin (2001, April 9). Novel insecticidal toxins from the venom of the spider Segestria florentina. Toxicon, 40, 125-130.