Team:NRP-UEA-Norwich/Results/Prebiotic
RESULTS - PREBIOTIC
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Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be.
Figure 1: A model structure of glycogen with acyl groups added.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
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Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
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Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
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Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
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Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
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Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
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Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Making constructs to express putative acyl-transferases in plant chloroplasts
AIM:
The aim of our prebiotic is to produce acylated/butrylated starch in plants. Methods to chemically acylate starch purified from plants already exist, but as they use strong chemicals and require heating they are not environmentally friendly. Using various acyltransferases, we hope to acylate starch in plants. We’ll be using a model plant, Nicotinia benthamiana, for initial tests because we can get results within a few days. Later on we’d aim to make transgenic plants in a species makes a lot of starch such as maize (corn), potatoes or wheat.
METHOD:
We used Golden Gate Cloning and the Plant Standard Syntax1 to make our constructs. We used 35s promoter from Cauliflower Mosiac Virus (BBa_K1467101), to drive constitutive expression. We made a chloroplast transit peptide (New part - BBa_K1618028) in an N-terminal translation fusion with the acyl transferases for the protein to reach the chloroplast, where starch is produced.
In order to confirm that the transit peptide was functional we made a second set of constructs that had a fluorescent reporter in a C-terminal translational fusion.
To deliver the constructs to plants we needed to use Agrobacterium tumefaciens as a shuttle chassis. This meant parts needed to be assembled into a binary vector with origins of replication for both E.coli and A. tumefaciens. In order to submit the parts to the registry we also assembled parts into the MoCloFlipper pSB1C3 that accepts Golden Gate Parts into the pSB1C3 backbone.
All cloning was done according to the Golden Gate one-step Digestion-Ligation Protocol provided on our protocols page here.
Results:
Successful assembly into the binary vector.
Putative clones were screened by colony PCR (see Protocols for method - here) using primers that flank the insertion sites (Figure 1).
Figure 1: The gel lanes are as follows – Ladder, 1&2 = MAO, 3&4 = MAO + YFP 5&6 = RV3034c, 7&8 = RV3034c + YFP, 9&10 = RV3037c, 11&12 = RV3037c + YFP, 13&14 = RV3030, 15&16 = RV3030 + YFP, ladder. The gel image above indicated that the complete transcriptional units were successfully cloned.
A single colony for each construct was mini-prepped and sequenced before progression to transformation into A. tumefaciens.
Successful assembly into pSB1C3.
Putative clones were miniprepped (see Protocols for method - here) and screened by digestion with NotI (Figure 2) and also with EcoRI/PstI (Figure 3) to confirm that cloning was successful and that our constructs had no internal BioBrick restriction sites.
Figure 2: Gel lanes are as follows - Ladder, 1&2 = MAO, 3&4 = MAO + YFP, 5&6 = RV3034c, 7&8 = RV3034c + YFP, 9&10 = RV3037c, 11&12 = RV3037c + YFP, 13&14 = RV3030, 15&16 =RV3030 + YFP, Ladder. These indicate that there are no unwanted internal Not1 restriction sites within our constructs.
Figure 3: Gel lanes are as follows - Ladder, 1&2 MAO + GFP, 3&4 = RV3034c + GFP, 5&6 = RV3037c + GFP, 7&8 = RV3030 + GFP, 9&10 = MAO + GFP, 11&12 = RV3034c + GFP, 13&14 = RV3037c + GFP, 15&16 = RV3030 + GFP. Lanes 1-8 were digested with Not1 while lanes 9-16 were digested with EcoR1/Pst1. The results indicate that the new tags allow our constructs to be BioBrick compatible.
A single colony for each construct was mini-prepped and sequenced before shipping to the Registry.
References
1. Patron et al (2015) Standards for Plant Synthetic Biology: A Common Syntax for Exchange of DNA Parts doi: 10.1111/nph.13532/full
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Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
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Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
This new property means that silicon-based light detectors identify spin, so more information can be. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin. Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.Currently, copper wires transfer information in a computer; the process is slowed down as the wires heat up. "Photonic" and "spintronic" computing is the principle of transferring information by light or electron spin.
