Part:BBa_K1415002

PBAN (Mamestra brassicae)

Introduction:PBAN (Pheromone Biosynthesis Activating Neuropeptide)

Fig.1-1 A coding gene of a Spodoptera litura'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 Spodoptera litura's PBAN.
See our expanding PBAN(SL) parts collection: Pcons+B0034+PBAN(Spodoptera litura) and Pcons+B0034+PBAN(Spodoptera litura)+B0034+BFP+J61048

Fig.1-2 Working mechanism of PBAN

Target insect:Oriental Leafworm Moth (Spodoptera litura)

Fig.1-1 A coding gene of a Spodoptera litura's PBAN

The experiment of PBAN

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.1-4 showed the correct size of the PBAN, and proved that we successful ligated the PBAN DNA sequence onto an ideal backbone.

Fig.1-4 The PCR result of the 9 different kinds of PBAN. The DNA sequence length of PBANs are around 100～150 bp, so the PCR products should appear at 415～515 bp.

Below are biobrick serial numbers of PBAN abbrevation:

BM: BBa_K1415001　　　　MB: BBa_K1415002　　　　AI: BBa_K1415003

LD: BBa_K1415004　　　　SL: BBa_K1415005　　　　HAH:BBa_K1415006

AS: BBa_K1415007　　　　SI: BBa_K1415008　　　　AA: BBa_K1415009

Application of the part

The experiment of P_cons+RBS+PBAN-SL

Fig.2-1-6-1 Protein Electrophoresis of Pcons + RBS + 5 different kinds of PBAN (control: plasmid of Pcons+RBS) Each peptide of PBAN is an around 30 amino acids, so we can see the band of PBANs at 2~4 kDa.

Below are biobrick serial numbers of PBAN abbrevation:

BM: BBa_K1415001　　　AA: BBa_K1415009　　　LD: BBa_K1415104

AS: BBa_K1415007　　　SL: BBa_K1415005

Behavior of Target Insects After PBAN Treatment

　

The experiment of P_cons+RBS+PBAN-SL+RBS+BFP+Ter

Fig.3-4-1 Biobrick of Pcons + RBS + PBAN(SL) + BFP + Term.

Fig.2-1-7 The growth curve of E.coli containing Pcons + RBS + 9 different kinds of PBAN + RBS + BFP + Ter plasmid (control is the competent cells which can not emit blue light).

Fig.2-1-8 The blue light fluorescence expression curve of E.coli containing Pcons + RBS + 9 different kinds of PBAN + RBS + BFP + Ter plasmid (control is the competent cells which can not emit blue light).

Fig.2-1-5 Blue Fluorescence of Pcons + RBS + 9 different kinds of PBAN (control: E.coli containg Pcons+RBS Plasmid).　Below are biobrick serial numbers of PBAN abbrevation:</p> SL: BBa_K1415005　　　　BM: BBa_K1415001　　　　MB: BBa_K1415002

AI: BBa_K1415003　　　　LD: BBa_K1415004　　　　HAH:BBa_K1415006

AS: BBa_K1415007　　　　SI: BBa_K1415008　　　　AA: BBa_K1415009

modeling

Fig.3-4-2 Modeling result of Pcons + RBS + PBAN(SL) + BFP + Ter. The blue line is the expression profile of the theoretical biobrick. And the green line is the expression data of Pcons + RBS + PBAN(SL) + BFP + Ter. And the red line is the adjusting line from the green and blue one. This line represent the correcting line of theoretical data and real condition data which can make our model not only fit the theoretical condition but also stay away from experimental bias.

The device we design and working mechanism

Fig.1-2-7 Our Project Overview.