Difference between revisions of "Part:BBa K1415001"
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1415002 short</partinfo>
 
<partinfo>BBa_K1415002 short</partinfo>
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[[File:BM.png|right|300px| ]]
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<h1>'''Introduction:''' PBAN (Pheromone Biosynthesis Activating Neuropeptide)</h1>
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[[File:PCRBM6.png|right|200px| The PCR result of our 9 different kinds of PBAN. Because our PBAN DNA sequence length is around 100～150 bp., the PCR result should be 415～515 bp.]]
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<div>[[File:HAH.png|thumb|right|300px|'''Fig.1-1''' A coding gene of a (Helicoverpa armigera Hubner'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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https://parts.igem.org/Registry:Feature_requests
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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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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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<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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Target insect:Silkworm (Bombyx mori)
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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 Helicoverpa armigera Hubner's PBAN.'''
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<br>
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See our expanding PBAN(Helicoverpa armigera Hubner) parts collection:
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[https://parts.igem.org/Part:BBa_K1415106 Pcons+B0034+PBAN(Helicoverpa armigera Hubner)] and
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[https://parts.igem.org/Part:BBa_K1415206 Pcons+B0034+PBAN(Helicoverpa armigera Hubner)+B0034+BFP+J61048 ]
  
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Spread: the place where farmers want to feed.
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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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Characteristics : It is entirely dependent on humans for its reproduction and does not occur naturally in the wild.
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<h1>'''Target insect:''' Cotton bollworm (Helicoverpa armigera Hubner)</h1>
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[[File:HAH-insect.png|thumb|right|900px|'''Fig.2-1''' Introduction of Helicoverpa armigera Hubner]]
  
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Damage: None
 
  
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Control: Not necessary
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<h1>The experiment of PBAN</h1>
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Peptide Sequence： LSEDMPATPADQEMYQPDPEEMESRTRYFSPRL
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[[File:PCRHAH5.png|left|thumb|900px|<p style="padding: 10px !important;">'''Fig.2-2''' The PCR result of the PBAN-SL. 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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[[File:Bombyx mori.png|300px|link=|frameless|left]]
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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(Helicoverpa armigera Hubner)]]
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</div>
  
  

Revision as of 15:46, 17 October 2014

PBAN (Mamestra brassicae)

Introduction: PBAN (Pheromone Biosynthesis Activating Neuropeptide)

Fig.1-1 A coding gene of a (Helicoverpa armigera Hubner'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 Helicoverpa armigera Hubner's PBAN.
See our expanding PBAN(Helicoverpa armigera Hubner) parts collection: Pcons+B0034+PBAN(Helicoverpa armigera Hubner) and Pcons+B0034+PBAN(Helicoverpa armigera Hubner)+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 (Helicoverpa armigera Hubner)

Fig.2-1 Introduction of Helicoverpa armigera Hubner


The experiment of PBAN

Fig.2-2 The PCR result of the PBAN-SL. 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(Helicoverpa armigera Hubner)






Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 18
  • 1000
    COMPATIBLE WITH RFC[1000]