What is it about?
Linkers are short polypeptide sequences that connect proteins in protein fusions, analogous to the couplings that hold freight cars together on a train. This paper describes a straightforward method for cloning multiple linkers into a single protein fusion and the subsequent evaluation of linker impacts on protein expression and activity.
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Why is it important?
Linkers are known to be important for the optimal function of protein fusions, but there are relatively few straightforward methods for their incorporation and evaluation. This straightforward method can be used to insert linker sequences that vary by both length and sequence into genes encoding protein fusions.
Perspectives
This work was inspired by two observations 1) the lack of a method for the iterative evaluation of the impacts of linkers on protein fusion activity and expression and 2) the ability of PCR primers with repetitive sequence content to anneal promiscuously and generate amplicons that were longer than originally intended. Where other published methods require dozens of primers for linker incorporation and evaluation, protaTETHER requires only a handful of PCR primers (minimum of 4) to accomplish the same result. We are happy consider collaborations if you're interested in trying the protaTETHER method on your protein fusion of choice.
Robert Hughes
East Carolina University
Read the Original
This page is a summary of: protaTETHER - a method for the incorporation of variable linkers in protein fusions reveals impacts of linker flexibility in a PKAc-GFP fusion protein, FEBS Open Bio, April 2018, Wiley,
DOI: 10.1002/2211-5463.12414.
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