2 replies to this topic

[HOYAJM]

I inserted a gene of interest into a pET32a vector downstream of the T7 promoter. However, while doing this I also removed the RBS unintentionally. I plan to express the construct in BL21 E. coli to purify the protein of interest. My question is... will there be expression of the protein despite the lack of the RBS? Will the ribosomes be able to translate the mRNA without it? I am doubtful it can, but I am certainly trying to prevent having to make a new construct.


Thanks

Edited by HOYAJM, 09 February 2012 - 10:46 AM.

[scolix]

I am not sure how it works with prokaryotic expression. But in eukaryotes, there can be reduction in the translation if RBS is lacking.

[phage434]

It might express, but I would not count on it. An experiment will tell you, or you can fix it.