Yes, definitely. This way you get the gen into bac. subt. Sal1 has to be in the vector too, but I think one of their vectors will have.
But:
The lux operon is from gram negative bacteria, and b.s. is a gram positive. It may glow weaker (or even not at all. )
But if you have access to the lab, and can use the chemicals for free, I'd try it ;) And the distributor too may give you the plasmid for free.
Those plasmids don't cut out the resistance. So it's not wise to handle them without care so they escape into the wilderness. Although, b.s. is quite the opposite of a pathogen. Just make sure to take the same care you should with each bacterium.
Best, choose a resistance that's very common in natural bacteria. I read an organization saying "Kanamycin resistance is very wide-spread in wild bacteria. So it's the best bet for use in genetically modified crops" (So if you make the bacilli resistant to kanamycin and if one escapes into the wilderness, it's still better than chloramphenicol resistant etc. )
That source was very helpful, I had never heard of B.Subtilis, seems like a great bacteria. I think I would definitely choose one of their plasmids. Since their plasmids already have resistance, could I make the construct by just cutting pVIB and my vector of choice with Sal1, ligating together, then transforming into B.Subtilis?
On Sunday, July 22, 2012 2:08:51 PM UTC-4, Mega wrote:> Also how exactly would you build a plasmid that would insert part of its genes into the chromosome, any certain plasmids need to be used?
http://www.bgsc.org/Catpart4.pdf
Here are some plasmids listed. It is written that on request, they may send you a plasmid for free if you use it for non-profit things.
Well, the plasmid needs two flanking sites that fit to a gene in the Bac. Subt. Chromosome. Then in 1 out of 10000 cases, homologous recombination occurs. The plasmid does not replicate, so if the bacillus doesn't die from the antibiotic and forms a colony, it has the gene casette integrated.
In theory, you could also use pUC19, fit in flanking sites of B.Subt. (where to get the template?, can you design the primers?), inside the flanking sites Lux operon + Resistance , amplify it in E.coli, transform B.S.
> I will have full time access to my schools laboratory.
Awsome ;)
> For that project would you recommend B.Subtilis?
Yeah, because of the spore formation. E.Coli will die on the plates after a while when the sugar is out. After at last one year, you cannot revive them again.
The spores of B.S., when they 'smell' better conditions, will 'wake up' again and in theory, still after 50 years of dryness, etc. you'll get a glowing B.Subt. again.
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