Do you know of any website which would predict RNAse E cutting site? or how to predict it?
We are having problems expressing a foreign mRNA in E.coli and one possibility was that the mRNA was being degraded very quickly.
RNase E cleaves AU-rich single stranded RNA, and is responsible for general mRNA decay. In general, the half-lives of E.coli mRNAs is very short (1~2min). Faster decay of your mRNA by specific RNase E attack might not be the top reason for your problem. Do you have a strong/constitutive promoter? and good SD/ATG?
the gene is hat could lder the T7 promoter. what does SD here refers to? and yes I do have a start codon in the construct. What does this tells?
I am wrapping up my thesis. So I am just going to summarise all the probable reasons that could lead to my failed expression in nut shell.
SD: Shine Dalgarno
Inefficient translation could be a likely reason for instability because the mRNA is not well protected by translating ribosomes.
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