Difference between revisions of "Part:BBa K1415003"
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1415003 short</partinfo>
 
<partinfo>BBa_K1415003 short</partinfo>
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[[File:AI.png|800px|link=|frameless|right]]
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<h1>'''Introduction:''' PBAN (Pheromone Biosynthesis Activating Neuropeptide)</h1>
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[[File:PCRAI5.png|200px|link=|frameless|right]]
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<div>[[File:HAH.png|thumb|right|300px|'''Fig.1-1''' A coding gene of a (Agrotis ipsilon's PBAN ]]</div>
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<p style="font-size:120%">'''Mechanism of PBAN'''</p>
 
PBAN (Pheromone Biosynthesis Activating Neuropeptide) is one kind of peptides that can activate biosynthesis of pheromones of insects we target. Once a PBAN binds with the G-protein coupled receptor on an insect’s pheromone gland, the signal send by the G-protein coupled receptor activates the kinase and phosphatase, and then kinase and phosphatase can activate enzymes that participate in the biosynthesis of insect pheromone, which will be emitted.
 
PBAN (Pheromone Biosynthesis Activating Neuropeptide) is one kind of peptides that can activate biosynthesis of pheromones of insects we target. Once a PBAN binds with the G-protein coupled receptor on an insect’s pheromone gland, the signal send by the G-protein coupled receptor activates the kinase and phosphatase, and then kinase and phosphatase can activate enzymes that participate in the biosynthesis of insect pheromone, which will be emitted.
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<br><br>
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<p style="font-size:120%">'''Features of PBAN'''</p>
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'''1. Species-specific''': PBAN is species-specific just like pheromones, meaning that every kind of insect produces specific PBAN that only binds with its specific receptor, resulting in the production of a particular pheromone.
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<br><br>
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'''2. Small and simple:''' The coding sequence for a PBAN is only around 100 base pairs. For E.coli, 100 base pairs is totally within its working capacity. Therefore, E.coli can be a low-cost PBAN factory. By transforming the DNA sequences for different PBAN into the E.coil, we can even gain a variety of PBANs. 　
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<br><br>
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'''3. Secreted directly:''' Because PBAN is secreted by the insect itself, the insect would not form a resistance to it compare to use pesticide.
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<br><br>
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Together, using PBAN is totally a environmental friendly way for solving harmful insects problems with easily triggering pheromone production by contacting with its receptor.
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<br><br>
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'''This part is a coding gene of a Agrotis ipsilonr's PBAN.'''
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<br>
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See our expanding PBAN(Agrotis ipsilonr) parts collection:
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[https://parts.igem.org/Part:BBa_K1415103 Pcons+B0034+PBAN(Agrotis ipsilon)] and
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[https://parts.igem.org/Part:BBa_K1415203 Pcons+B0034+PBAN(Agrotis ipsilon)+B0034+BFP+J61048 ]
  
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So,if we ligate the constitutive promoter and ribosome binding site,we can make our E.coli produce our special peptides constantly.
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[[File:2014NCTUGprotein.jpg|800px|thumb|center|'''Fig.1-2''' Working mechanism of PBAN ]]
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''Reference:<p>Ada Rafaeli, Pheromone biosynthesis activating neuropeptide (PBAN): Regulatory role and mode of action, ELSEVIER, General and Comparative Endocrinology 162 (2009) 69–78.</p>''
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<br><br><br>
  
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Target insect:Black cutworm (Agrotis ipsilon)
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<h1>'''Target insect:''' Cotton bollworm (Agrotis ipsilon)</h1>
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[[File:HAH-insect.png|thumb|right|900px|'''Fig.2-1''' Introduction of Agrotis ipsilonr]]
  
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Spread: This Caterpillar can be found, as various species, throughout North America.
 
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Characteristics: Cutworms common in Georgia fields are black (Agrotis ipsilon (Ashmed)), granulate (Agrotis subterranea (Fabricius)) and variegated cutworms (Peridroma saucia(Hubner)). These are moths in the family Noctuidae. Full-grown cutworm larvae are 1.5 to 2 inches long. Coloration will vary among species, but all tend to be stout-bodied caterpillars with four sets of prolegs. They have the tendency to curl into a ball when disturbed.
 
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Damage: Almost any plant can be attacked in the seedling stage. Cotton and certain vegetables sometimes have stand reductions. Climbing cutworms, such as the granulate cutworm, can be serious foliage feeders on some crops such as peanuts.
 
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Control: Bacillus thuringiensis, a widely available caterpillar-killing bacterium,is a very effective control for climbing cutworms as well as for the surface feeders.
 
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Peptide Sequence： LADDTPATPADQEMYRPDPEQIDSRTKYFSPRL
 
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<br><br><br>
 
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[[File:Agrotis ipsilon.png|300px|link=|frameless|left]]
 
  
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<h1>The experiment of PBAN</h1>
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[[File:PCRHAH5.png|left|thumb|900px|<p style="padding: 10px !important;">'''Fig.2-2''' The PCR result of the PBAN-AI. The DNA sequence length of PBANs are around 100～150 bp, so the PCR products should appear at 415～515 bp.</p>]]
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<div style="display: block; height: 530pt;">
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After receiving the DNA sequences from the gene synthesis company, we recombined each PBAN gene to PSB1C3 backbones and conducted a PCR experiment to check the size of each of the PBANs. The DNA sequence length of the PBAN are around 100～150 bp. In this PCR experiment, the PBAN products size should be near at 415～515 bp. The '''Fig.2-2''' showed the correct size of the PBAN, and proved that we successful ligated the PBAN DNA sequence onto an ideal backbone.
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[[File:nctu006pcr.jpg|right|thumb|650px|'''Fig.2-3''' The plate of part-PBAN(Agrotis ipsilon)]]
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</div>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 15:59, 17 October 2014

PBAN (Agrotis ipsilon)

Introduction: PBAN (Pheromone Biosynthesis Activating Neuropeptide)

Fig.1-1 A coding gene of a (Agrotis ipsilon's PBAN

Mechanism of PBAN

PBAN (Pheromone Biosynthesis Activating Neuropeptide) is one kind of peptides that can activate biosynthesis of pheromones of insects we target. Once a PBAN binds with the G-protein coupled receptor on an insect’s pheromone gland, the signal send by the G-protein coupled receptor activates the kinase and phosphatase, and then kinase and phosphatase can activate enzymes that participate in the biosynthesis of insect pheromone, which will be emitted.

Features of PBAN

1. Species-specific: PBAN is species-specific just like pheromones, meaning that every kind of insect produces specific PBAN that only binds with its specific receptor, resulting in the production of a particular pheromone.

2. Small and simple: The coding sequence for a PBAN is only around 100 base pairs. For E.coli, 100 base pairs is totally within its working capacity. Therefore, E.coli can be a low-cost PBAN factory. By transforming the DNA sequences for different PBAN into the E.coil, we can even gain a variety of PBANs. 　

3. Secreted directly: Because PBAN is secreted by the insect itself, the insect would not form a resistance to it compare to use pesticide.

Together, using PBAN is totally a environmental friendly way for solving harmful insects problems with easily triggering pheromone production by contacting with its receptor.

This part is a coding gene of a Agrotis ipsilonr's PBAN.
See our expanding PBAN(Agrotis ipsilonr) parts collection: Pcons+B0034+PBAN(Agrotis ipsilon) and Pcons+B0034+PBAN(Agrotis ipsilon)+B0034+BFP+J61048

Fig.1-2 Working mechanism of PBAN
Reference:

Ada Rafaeli, Pheromone biosynthesis activating neuropeptide (PBAN): Regulatory role and mode of action, ELSEVIER, General and Comparative Endocrinology 162 (2009) 69–78.




Target insect: Cotton bollworm (Agrotis ipsilon)

Fig.2-1 Introduction of Agrotis ipsilonr


The experiment of PBAN

Fig.2-2 The PCR result of the PBAN-AI. The DNA sequence length of PBANs are around 100～150 bp, so the PCR products should appear at 415～515 bp.

After receiving the DNA sequences from the gene synthesis company, we recombined each PBAN gene to PSB1C3 backbones and conducted a PCR experiment to check the size of each of the PBANs. The DNA sequence length of the PBAN are around 100～150 bp. In this PCR experiment, the PBAN products size should be near at 415～515 bp. The Fig.2-2 showed the correct size of the PBAN, and proved that we successful ligated the PBAN DNA sequence onto an ideal backbone.

Fig.2-3 The plate of part-PBAN(Agrotis ipsilon)














Peptide Sequence： LADDTPATPADQEMYRPDPEQIDSRTKYFSPRL


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]