9000 turnovers per second = from Desulfovibrio vulgaris.

Glucose transporter: "

Trophic Conversion of an Obligate Photoautotrophic Organism Through Metabolic Engineering"

http://www.kazusa.or.jp/codon/ - "Count Codon" service

Inosine = universal base pair

Stratagene Mutazyme = good system for ePCR (less mutational bias - inherently error prone polymerase)

Functional proteins from a random-sequence library: completely random sequence of DNA, combined with mRNA display (?) allowed the generation of a novel ATP binding protein:O

Directed Evolution of Novel Protein Functions:
- From a theoretical perspective, the frequency of functional proteins in the entire protein sequence space is high enough that novel proteins could be generated simply by random methods (but still, low frequency). Pretty cool! If you can come up with a screening mechanism, then you can generate proteins that can do anything:O
- Recognising that it may take several sequential mutations to achieve a desired function, Chen and Zhao developed a screening methodology that involves iterative experiments that attempt to develop a function protein through a series of logical, intermediate steps
- To develop novel proteins, only a small number of mutations are necessary and these are normally localised to the active site

"GC clamp" - add a GC rich region next to sequence of interest. Use in primers to enhance binding to sequence (GC bonds = stronger)

Directed evolution of biocatalytic processes:
- Improvements in one trait may be offset by decline in another trait
- In order to generate useful industrial biocatalysts, screening conditions in directed evolution experiments should closely mimic those that will be found in the industrial setting. Can perform microscale experiments that can be scaled up to large bioreactors
- It's quite hard to ligate mutagenised DNA fragments into a plasmid to transform an E coli - ie you lost a fair bit of your mutation realm simply due to the inefficiency of ligation and transformation

Directed evolution of industrial enzymes
- 'you get what you screen for'