Hello everyone,
I would like to study the protein profile of TB, so I have to transfect DNA of M. TUBERCULOSIS with a type of cell line. Kindly your Opinions and recommendations.
Thank you
In PubMed there is this article "Establishment of RAW264.7 cell line stably expressing Mycobacterium tuberculosis protein ESAT-6" but unfortunately is in chinese. http://www.ncbi.nlm.nih.gov/pubmed/22117523.
Also, in english there is this interesting one from 2012:
Mycobacterial Shuttle Vectors Designed for High-Level Protein Expression in Infected Macrophages
(which I even attached to this message).
Please if helps, mark with green up arrow (upvote) for this answer. Thanks.
Hii,
It all depends upon what proteins you want to study. MTB proteomics has been studies extensively.
Instead of Transfecting, DNA, you can use THP-1 cell line and induce infection with MTB bacilli, provided you have a BSL facility Since problems with DNA is amount of expression in ex vivo system. after infecting the cells you could study host as well as MTB proteins and their expression. Possibly TLR induced proteins are better to investigate.To make study interesting you could use WCL fraction from different growth periods of MTB in liquid culture and study differences in expression of proteins in culture and in ex vivo system.
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