Protein Expression Vectors – Template, Codon Bias, Affinity Tags and Epitopes [Part 2]

References/Resources: https://www.patreon.com/the_Crux

All videos on Genetic Engineering: https://www.youtube.com/playlist?list=PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf

In this video, we explore how to obtain templates for protein expression vectors depending on the gene structure (intron and exon language). Prokaryotic genes are intron-less, whereas eukaryotic genes often have introns. Obtaining template for protein expression vectors is a challenge and not straightforward for eukaryotic hosts. Converting mRNA to cDNA to clone the templates becomes the way to obtain templates for eukaryotic genes, unless they are intron-less. For heterologous protein expression, a serious problem appears from codon bias when optimizing for efficient protein expression. Codon bias and its optimization is a host specific issue. We discuss some strategies and issues around codon usage optimization. Codon optimization is always a good thing because it can affect translation speed and efficiency and even interfere with protein folding. Finally, you could engineer the expressed proteins according to your requirements with purification tags, localization signals, and many other things. We look at some commonly used protein engineering and modifications along with their application and use.

Jump to your favorite section:
00:00 Template
00:30 Gene Structure
02:48 Getting the template
07:02 Codon Bias
09:30 Codon Bias Optimization
11:18 Extra Engineering
11:55 Localization Signal
12:48 Affinity Tags
14:38 Other Modifications

References/Resources: https://www.patreon.com/the_Crux

Suggested videos:
Protein Expression Vectors [Part 1]: https://youtu.be/3_WnvVgIkYo

*Must watch playlists*
All videos on Genetic Engineering: https://www.youtube.com/playlist?list=PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf
All videos on DNA Replication: https://youtube.com/playlist?list=PL0Ymnd-zt4IgLefDaeqicJbCBnbLqnDJ7
All videos on Transcription: https://www.youtube.com/playlist?list=PL0Ymnd-zt4Ij2VcAOHNUweftSElVsc-MX
All videos on Translation: https://www.youtube.com/playlist?list=PL0Ymnd-zt4IjAAAMlPLDIXbs692Yp14xa

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