Difference between revisions of "Part:BBa K1415004"
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<partinfo>BBa_K1415003 short</partinfo>
 
<partinfo>BBa_K1415003 short</partinfo>
  
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[[File:LD.png|300px|link=|frameless|right]]
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<h1>'''Introduction:''' PBAN (Pheromone Biosynthesis Activating Neuropeptide)</h1>
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[[File:PCRLD6.png|200px|link=|frameless|right]]
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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>
 
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 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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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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Target insect:Gypsy Moth (Lymantria dispar)
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Spread: It has a range which covers Europe, Africa, and North America.
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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:Gypsy moth caterpillars change appearance as they grow. Young caterpillars are black or brown and about ¼ inch (.6 cm) in length. As they grow, bumps develop along their backs along with coarse, black hairs. Each of the 11 sections of a developed caterpillar will have two coloured spots, the first five pairs, blue, and the last six, red. Mature caterpillars can be as long as 2 ½ inches (6.35 cm).
 
  
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Damage: It is classified as a pest, and its larvae consume the leaves of over 500 species of trees, shrubs and plants. The gypsy moth is one of the most destructive pests of hardwood trees in the eastern United States.he gypsy moth was considered a nuisance just ten years after their release. It included an account of all the trees being defoliated, caterpillars covering houses and sidewalks and that the caterpillars would rain down upon residents. The first outbreak occurred in 1889. An eradication program was begun in 1890.
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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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Control: Tanglefoot Pest Barrier or Sticky Tree Bands can be placed around tree trunks to help curtail the caterpillars movement into and out of the tree canopy. Apply Bacillus thuringiensis, var. kurstaki or Monterey Garden Insect Spray (Spinosad) to the leaves of trees to kill gypsy moth caterpillars.
 
  
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<h2>The experiment of PBAN</h2>
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[[File:PCRMB4.png|left|thumb|900px|<p style="padding: 10px !important;">'''Fig.2-2''' The PCR result of the PBAN-MB. 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:Lymantria dispar.png|300px|link=|frameless|left]]
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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.
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===Usage and Biology===
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<span class='h3bb'>Peptide Sequence： LADDTPATPADQEMYRPDPEQIDSRTKYFSPRL</span>
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<partinfo>BBa_K1415003 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K1415003 parameters</partinfo>
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Revision as of 15:54, 17 October 2014

PBAN (Agrotis ipsilon)

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 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 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-MB. 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.