Antisense phenomena is generally absent in bacteria. But u can knock down gene using Sigma targetron kit and is effective for M. tb. Another way is using suicidal vector. Clone gene of interest in suicidal vector and introduce in native strain. If the protein corresponding to gene of interest is prest in bacterial, the bacteria will survive else it will die.
The antisense technology in prokaryotes does not work like in eukaryotes although the production of a complete complementary RNA will decrease the translation of your target RNA; rRNA cannot translate dsRNA.
My advice is to produce a complementary mRNA for your target gene, here an interesting article about that:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1716169/
Hi Silvia
I agree with Manuel
I successfully demonstrated the antisense tecnology in Mtb
plz look into the following link...
http://www.ncbi.nlm.nih.gov/pubmed/20864475
Gud Luck
Great! Ive read your paper and it's very interesting. I would like to use this vector (pMV361) but I don't know where I could find it. Do you know where can I get it? I'm working in Peru and I would like to introduce this kind of antisense technology applied in M. smegmatis or M. tuberculosis.
Thank you a lot.
Thanks Silvia
Actually I left my PhD lab where I used this vector but You can always get from company Like addgene....
plz look at this link http://www.addgene.org/29462/
In this vector you will find additional sequences like ccdB and CAT insert, which You can easily excise to get pMV361.
I hope this will help.
Gud Luck
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