Elaboration on Teacher's Project: Fast Gene Expression and Detection of a Bacteria's presenceTeacher had suggested we come out with a way for bacteria to rapidly Express a gene, so we can rapidly detect the presence of the bacteria in question and i guess I would blog on some ways bacteria can rapidly Express a Gene.
Using IGEM BioBrick Parts: By Optimizing Promoters, Ribosome Binding Sites and changing the DNA code, we could make our bacteria express a Gene Rapidly.- Use a High Copy plasmid , Eg pMB1 , which leads to quick expression of a protein.
- Use a strong promoter , to promote strong binding of a Gene. A strong promoter would have high POPs or polymerase (transcription activity) per second,
- Codon optimization. Where you use Codons , where you have a higher tRNA pool for. Read Codon usage bias or Codon Degeneracy(Multiple codons for one Amino acid) , I am not sure if a high GC content which would make the RNA more stable help increase DNA expression
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| E. coli has more tRNA for CGC than CGG so using CGC will increase gene expression :source Biologics Corp |
Other Ideas we can explore include:
- Using natural Selection to select for a Bacteria that will express the Gene Fast.
- Using a better bacteria Chassis, eg a Lactobacillus that grows fast, We could explore Yeast, which if it works would be the battle 75% won, The Yeast in instant Yeast work well in multiplying rapidly, Might be be cause there is already a high cell density or some other reason; Needs investigation!!!!!
- Using Super Stable mRNA that has a long lifespan, this is not likely to work, as mRNAs are inherently unstable but we could explore using mRNA that has a long lifespan . This way the bacteria grows and produces mRNA , that does not degrade and thus expression of a gene just increases .
- Design a bacteria to get rid of RNAse which degrades RNA, i skimed though this paper for the idea. As getting rid of RNAse would likely kill the bactria eventually, we could activate the Gene for deactivating RNAse or knocking our RNAse, crispr? upon detection of IPTG or Lactose.
- Using Multiple Plasmids with Diffrent origin of replication, this may work but it will come at the cost of increased metabolic Strain during bacteria Multiplication.
- Use Antibiotic resistance? , get our bacteria to express an antibiotic to kill off competing bacteria so as to grow fast and furious. This way we can rapidly detect the bacteria's pressance.
- Work on my Bacteria USB3.0 Idea where we have a bacteria1 that can only produce AHL or a Biosignaling molecule. Students can freely play with the bacteria1, and upon a need to detect the bacteria you can add a small sample of the bacteria to a secondary culture medium full of bacteria2, upon detection of bacteria1's biosignalling molecule the bacteria2 will produce an output say a color . This Idea tho may be suceptible to False positives , if there are other bacteria with a similar biosignaling molecule If sucessful we could extend the idea and produce diffrent variations of bacteria1 with diffrent biosignalling molecules and Bacteria2 will be able to give a diffrent output based on the diffrent biosingaling molecules
- Supplement Bacteria with Iron, to make them grow faster, :\
