How can we make the proteins using Cell-free display, starting with mRNA directly?

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Catherine Vaquero

What sort of cell free system do you use and what sort of mRNA (purified, total RNA, polyA+RNA ???. Does your protein have a signal peptide??

The folding of a protein takes place while the translation progresses from the N terminus to the C terminus of the protein.

Jitendra Pant

Hello Catherine,

Thanks for replying. I want to know theoretically how is it possible to make protein without using cells but using mRNA? Regarding mRNA, its total RNA, and yes it has a signal peptide.

Please also forward if you have simple and good papers on the same.

regards and thanks

Jitendra

You have to prepare extra good total RNA preparation and remember that polyA+ RNA represents around 2 % of total RNA. Then add your RNA preparation to an in vitro tranlastion system and if your specific mRNA is sufficiently represented and if there is no degradation in your RNA preparation, you might hope to produce sufficient protein that you could detect with a specific AB among all proteins translated. You have to look in the web for recent review on protein translation and translationale in vitro system.

I am no more in the field of in vitro translation but I have translated mRNA in vitro and in different contexts during many years.

Thanks for your kind reply. Your suggestions are very helpful.

regards