Difference between revisions of "Part:BBa K1974020"
 
(6 intermediate revisions by 3 users not shown)
Line 4: Line 4:
 
<h1>'''Introduction:'''</h1>
 
<h1>'''Introduction:'''</h1>
 
[[File:2016_NCTU-FORMOSA_L.png|800px|thumb|center|'''Figure 1.'''snowdrop lectin+linker+6xHistag]]
 
[[File:2016_NCTU-FORMOSA_L.png|800px|thumb|center|'''Figure 1.'''snowdrop lectin+linker+6xHistag]]
−
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;According to reference, snowdrop-lectin is resistant to high temperature and would not be degraded by digestive juice. The species-specificity is based on the toxin, and the snowdrop lectin is the role of the carrier. When it is ligated by Pantide, which can increase the oral activity of Pantide.<br>
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Snowdrop (Galanthus nivalis) lectin is a protein that has special affinity to the glycoprotein on the surface of insect epithelial cells. According to reference, snowdrop-lectin is resistant to high temperature and would not be degraded by digestive juice. The species-specificity is based on the toxin, and the snowdrop lectin is the role of the carrier. When it is ligated by Pantide, which can increase the oral activity of Pantide.<br>
−
[[File:Lectin-basic.png|200px|thumb|center|'''Figure 2.'''. Snowdrop lection
+
 
 +
<h1>''' GreatBay-SCIE 2024'''</h1>
 +
This year, GreatBay-SCIE intergrate Galanthus nivalis agglutinin (GNA) into our experiment design. GNA is used for enhancing the oral and contact toxicity of the expressed mite and spider venom peptides (MVPs/SVPs). We constructed the plasmid pET28a-G1M5-His-SUMO-MVP/SVP-GNA-His (the MVPs can be PpVP2S, PpVP1S, or PpVP1F; the SVPs can be rCtx-4, Cs1A or HxTx-Hv1h). According to the toxicity bioassay, the MVP/SVP-GNA fusion proteins exhibit high contact efficacy. PpVP2S, a novel MVP we discovered by ourselves through conducting gene mining, achieves a spider mite mortality rate of 100% in 48 hours.
 +
<center><html><img src="https://static.igem.wiki/teams/5184/parts/vp-lethality-v.webp" width="600"/></html></center>
 +
<center><b>Fig2: A. Survival plot of 6 venom peptides against female ''T. urticae'' using a spraying method, CK is induced liquid culture of BL21(DE3), of which acts as control D. Lethality data of 6 venom peptides over 24, 48, and 72 hours, CK is induced liquid culture of BL21(DE3), of which acts as control; data is the means of ± SD of three parallel replicate experiments</b></center>
 +
 
 +
[[File:Lectin-basic.png|200px|thumb|center|'''Figure 3.'''. Snowdrop lection
 
The DNA sequence length of Snowdrop lectin is around 300-400 b.p. In this PCR experiment, the product’s size should be close to 550-700 b.p.]]
 
The DNA sequence length of Snowdrop lectin is around 300-400 b.p. In this PCR experiment, the product’s size should be close to 550-700 b.p.]]
−
[[File:2016NCTU_FORMOSA_mechanism.png|800px|thumb|center|'''Figure 3.''' Mechanism: In the alimentary canal, lectin binds to gut epithelial glycoproteins, and crosses the epithelium cell by endocytosis and exocytosis, and transported into haemolymph.]]
+
[[File:2016NCTU_FORMOSA_HL.png|200px|thumb|center|'''Figure 4.''']]
 +
[[File:2016NCTU_FORMOSA_SL.png|200px|thumb|center|'''Figure 5.''']]
 +
[[File:2016NCTU_FORMOSA_OL.png|200px|thumb|center|'''Figure 6.''']]
 +
[[File:2016NCTU_FORMOSA_mechanism.png|800px|thumb|center|'''Figure 7.''' Mechanism: In the alimentary canal, lectin binds to gut epithelial glycoproteins, and crosses the epithelium cell by endocytosis and exocytosis, and transported into haemolymph.]]
  
 +
<h1>'''Reference'''</h1>
 +
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
 +
<br>
 +
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,”
  
 
<!-- -->
 
<!-- -->

Latest revision as of 13:22, 2 October 2024


Snowdrop lectin+linker+6X His-Tag

Introduction:

Figure 1.snowdrop lectin+linker+6xHistag

      Snowdrop (Galanthus nivalis) lectin is a protein that has special affinity to the glycoprotein on the surface of insect epithelial cells. According to reference, snowdrop-lectin is resistant to high temperature and would not be degraded by digestive juice. The species-specificity is based on the toxin, and the snowdrop lectin is the role of the carrier. When it is ligated by Pantide, which can increase the oral activity of Pantide.

GreatBay-SCIE 2024

This year, GreatBay-SCIE intergrate Galanthus nivalis agglutinin (GNA) into our experiment design. GNA is used for enhancing the oral and contact toxicity of the expressed mite and spider venom peptides (MVPs/SVPs). We constructed the plasmid pET28a-G1M5-His-SUMO-MVP/SVP-GNA-His (the MVPs can be PpVP2S, PpVP1S, or PpVP1F; the SVPs can be rCtx-4, Cs1A or HxTx-Hv1h). According to the toxicity bioassay, the MVP/SVP-GNA fusion proteins exhibit high contact efficacy. PpVP2S, a novel MVP we discovered by ourselves through conducting gene mining, achieves a spider mite mortality rate of 100% in 48 hours.

Fig2: A. Survival plot of 6 venom peptides against female T. urticae using a spraying method, CK is induced liquid culture of BL21(DE3), of which acts as control D. Lethality data of 6 venom peptides over 24, 48, and 72 hours, CK is induced liquid culture of BL21(DE3), of which acts as control; data is the means of ± SD of three parallel replicate experiments
Figure 3.. Snowdrop lection The DNA sequence length of Snowdrop lectin is around 300-400 b.p. In this PCR experiment, the product’s size should be close to 550-700 b.p.
Figure 4.
Figure 5.
Figure 6.
Figure 7. Mechanism: In the alimentary canal, lectin binds to gut epithelial glycoproteins, and crosses the epithelium cell by endocytosis and exocytosis, and transported into haemolymph.

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,”

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]