Difference between revisions of "Part:BBa K1415009"
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<partinfo>BBa_K1415009 short</partinfo>
 
<partinfo>BBa_K1415009 short</partinfo>
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[[File:AAA.png|thumb|right|200px|'''Fig.1-1''' The biobrick of PBAN.]]
 
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[[File:PCRAA5.png|thumb|right|200px|'''Fig.1-2''' The PCR result of the PBAN(AA). The DNA sequence length of PBANs are around 100～150 bp, so the PCR products should appear at 415～515 bp ]]
 
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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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<h1>'''Introduction:''' PBAN (Pheromone Biosynthesis Activating Neuropeptide)</h1>
  
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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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<div>[[File:MB.png|thumb|right|300px|'''Fig.1-1''' A coding gene of a Mamestra brassicae's PBAN ]]</div>
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<p style="font-size:120%">'''Mechanism of PBAN'''</p>
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Target insect:Yellow fever mosquito (Aedes aegypyi)
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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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Spread:The yellow fever mosquito, Aedes aegypti, is a mosquito that can spread the dengue fever, chikungunya, and yellow fever viruses, and other diseases. The mosquito can be recognized by white markings on its legs and a marking in the form of a lyre on the thorax. The mosquito originated in Africa, but is now found in tropical and subtropical regions throughout the world.
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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 it's targeted receptor, resulting in the production of a particular pheromone.
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Characteristics: The mosquito can be recognized by white markings on its legs and a marking in the form of a lyre on the thorax.
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'''2. Small Simple Peptide:''' The coding sequence for a PBAN is only around 100 base pairs. To E.coli 100 base pairs is totally within its working capacity. And therefore, E.coli can be our low-cost PBAN factory. By synthesizing the DNA sequences for different PBAN into our factory, we can even produce a variety of PBANs. In addition, this factory is totally environmental friendly, unlike any pesticide we have seen.　
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Damage: The yellow fever mosquito, Aedes aegypti, is a mosquito that can spread the dengue fever, chikungunya, and yellow fever viruses, and other diseases.
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<br>
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'''3. Insects' own secretion:''' Because PBAN is a insect's own secretion, insects could not form resistance it. In addition, it can easily trigger pheromone production by coming in contact with its receptor.
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Control:(1) Empty water from containers such as flower pots, birdbaths, pet water dishes, cans, gutters, tires and buckets regularly to disrupt the mosquito breeding cycle. (2)Consider using an insect repellent, be sure to follow the label directions for applying the repellent. For help selecting a mosquito repellent, try our Insect Repellent Locator.
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<br><br>
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'''This part is a coding gene of a Mamestra brassicae's PBAN.'''
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<br>
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See our expanding PBAN(Mamestra brassicae) parts collection:
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[https://parts.igem.org/Part:BBa_K1415102 Pcons+B0034+PBAN(Mamestra brassicae)] and
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[https://parts.igem.org/Part:BBa_K1415202 Pcons+B0034+PBAN(Mamestra brassicae)+B0034+BFP+J61048 ]
  
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Peptide Sequence： DASSSNENNSRPPFAPRL
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[[File:2014NCTUGprotein.jpg|800px|thumb|center|'''Fig.1-2''' Working mechanism of PBAN ]]
  
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[[File:pestAA.png|thumb|left|300px|'''Fig.1-3'''  Targeted insect:Yellow fever mosquito. ]]
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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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<!-- Add more about the biology of this part here
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<br><br><br>
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===Usage and Biology===
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<h1>'''Target insect:''' Cabbage Moth (Mamestra brassicae)</h1>
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[[File:MB-insect.png|thumb|right|900px|'''Fig.2-1''' Introduction of Mamestra brassicae]]
  
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<partinfo>BBa_K1415009 SequenceAndFeatures</partinfo>
 
  
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<!-- Uncomment this to enable Functional Parameter display
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<h2>The experiment of PBAN</h2>
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===Functional Parameters===
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[[File:PCRMB4.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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<partinfo>BBa_K1415009 parameters</partinfo>
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<div style="display: block; height: 520pt;">
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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.

Revision as of 15:25, 17 October 2014

PBAN (Aedes aegypti)

Introduction: PBAN (Pheromone Biosynthesis Activating Neuropeptide)

Fig.1-1 A coding gene of a Mamestra brassicae'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 it's targeted receptor, resulting in the production of a particular pheromone.
2. Small Simple Peptide: The coding sequence for a PBAN is only around 100 base pairs. To E.coli 100 base pairs is totally within its working capacity. And therefore, E.coli can be our low-cost PBAN factory. By synthesizing the DNA sequences for different PBAN into our factory, we can even produce a variety of PBANs. In addition, this factory is totally environmental friendly, unlike any pesticide we have seen.　
3. Insects' own secretion: Because PBAN is a insect's own secretion, insects could not form resistance it. In addition, it can easily trigger pheromone production by coming in contact with its receptor.

This part is a coding gene of a Mamestra brassicae's PBAN.
See our expanding PBAN(Mamestra brassicae) parts collection: Pcons+B0034+PBAN(Mamestra brassicae) and Pcons+B0034+PBAN(Mamestra brassicae)+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: Cabbage Moth (Mamestra brassicae)

Fig.2-1 Introduction of Mamestra brassicae


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.