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Critirea for choosing 5' and 3' flanking sequences of yeast tRNA

tRNA cloning sequence

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#1 Balakumaran

Balakumaran

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Posted 03 August 2012 - 09:04 AM

I am working on enhancing recombinant protein productivity in yeasts through translation engineering.

With reference to this we have I have the following queries:

1. We would like to clone yeast tRNAs in a shuttle vector under the control of a strong promoter. From several publications we understood the role of 5' and 3' flanking sequences for efficient expression of tRNAs. In our study, we have chosen 40 bp upstream (5' TBP binding site) and 40 bp downstream (3' Termination site) of the mature tRNA sequence (from tRNA genome database). Is this selection procedure correct or is there any specific critirea for choosing the 5' and 3' flanking sequence of yeast tRNA?

2. Which online software tool is reliable for confirming the TBP binding site, termination site of tRNA sequences?

3. Which technique is used for analyzing functional expression of tRNAs




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