Difference between revisions of "Part:BBa K4140008"

Revision as of 07:59, 3 October 2022

lacZ alpha

Part Description

The LacZ-alpha fragment is encoded by this region and is derived from the pUC19 cloning vector. Because it is smaller than the LacZ-omega region, the LacZ-alpha fragment can be easily incorporated into a plasmid when the two non-functional LacZ gene fragments (alpha and omega) are co-expressed. X-gal (5-bromo-4-chloro-3-indoyl-d-galactopyranoside), a soluble colourless molecule that is a substrate of ß-galactosidase and generates a blue product when cleaved, is used to detect the alpha-complementation

Usage

alpha domain of ß-galactosidase (our reporter protein) which is produced in response to high levels of phenylalanine and generates a blue product when X-gal is cleaved which is a soluble colorless molecule that is a substrate of ß-galactosidase and generates a blue product when cleaved, is used to detect the alpha-complementation

Characterization of Mutational Landscape

After creating a multiple sequence alignment of the protein sequence and predicting mutational landscapes, the effect of these mutations on the evolutionary fitness of the protein is tested. The prediction of the mutational landscape by saturation mutagenesis of the LacZ protein. The (W77S ) mutation, as depicted in the chart, had the greatest score when compared to other mutations. On the other hand, it's clear that the (Y59L) had the least evolutionary fitness for LacZ protein. As displayed in Figure(1)

Figure(1) shows the mutational landscape of the LacZ protein

References

Sequence and Features