While the art of reading hand-poured sequencing gels is forgotten as the next generation sequencers spit out genome after genome, protein expression still often requires sweat, tears, and a lot of luck. I think this happens not because there is something different about protein expression in comparison with sequencing, but because people do not think about protein expression as an engineering challenge.
But where there is engineering, there should be a manual, right? Neither the literature reviews, which are full of confusing technical details, nor the protocols, which tell you how to do something but not why, fulfill this function.
What you need is something in between, a simple and maybe even funny introduction into protein expression, how to select parts, and optimize production. The good news is that our site is launching The Manual – an eBook called “The Bitesize Bio Guide to Protein Expression”. Written by a team of hands-on molecular biologists: an experienced postdoc and prolific BsB writer (me) and edited by an excellent scientific editor Emily Crow.
Check out the e-book content:
Chapter 1 Introduction
Chapter 2 Choosing an expression system
What is an expression system anyway?
Choosing your heterologous expression system
A horse called Escherichia
There is more to E. coli than BL21: Strain choice
Other parameters that you have at your disposal
Fifty shades of yeast
A very friendly armyworm
Insect cell lines
Vectors for expressing heterologous proteins in insect cells
Popular (cell) culture
I want to be (cell) free
Why would you use cell extracts?
Chapter 3 Exogenous protein expression: the problems
Why you need to know basic facts about your protein
Coding 101: Codon optimization 26
Are you content with your gene content?
Repeat sequences: Lego blocks of secondary structure
The game of introns
The importance of being soluble
The art of biological origami, or: Consider folding
Mess with protein sequence at your peril
Not so fast: How speed of expression can screw up your protein folding
But I couldn’t possibly fold without my chaperone!
Why won’t my freaky protein fold?
Return to sender: Expressing a protein in the wrong compartment
Export your protein
What if the expressed protein is toxic?
Pick up your copy of the eBook “The Bitesize Bio Guide to Protein Expression” today.
And if you want to know more about protein expression watch this webinar where I discuss the ins and outs of protein expression.