Revision as of 17:58, 16 October 2014

PBAN (Spodoptera litura)

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)

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.1-3 showed the correct size of the PBAN, and proved that we successful ligated the PBAN DNA sequence onto an ideal backbone.

Fig.1-3 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　　　　　　　　　

Fig.1-ˋ Protein Electrophoresis of Pcons + RBS + 4 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:

AI: BBa_K1415003　　　MB: BBa_K1415002　　　HAH:BBa_K1415006　　　SI: BBa_K1415008

Behavior of Target Insects After PBAN Treatment

Fig.2-3-1 Negative Control: Female moth without eating PBAN (Number = 0). Experiment: Female moth eating our PBAN (Number = 11). In this picture, we can see the PBAN effect that the female moth eating PBAN solution can release much sex pheromone, and attract many male moths.

Fig.2-3-2 Negative Control：sucrose solution, Experiment：Female moth eating PBAN solution. Also, we can see the PBAN effect again from this picture.

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

modeling

The device we design and working mechanism

modeling of device Sequence and Features