Imperial College London
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Professor Tony Cass

Faculty of Natural Sciences, Department of Chemistry

Senior Research Investigator
 
 
 
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Contact

 

+44 (0)20 7594 5195t.cass

 
 
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Location

 

301KMolecular Sciences Research HubWhite City Campus

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Summary

 

Publications

Citation

BibTex format

@article{Huang:2006:10.1002/biot.200600091,
author = {Huang, X and Zhang, X-E and Zhou, Y-F and Zhang, Z-P and Cass, AEG},
doi = {10.1002/biot.200600091},
journal = {Biotechnol J},
pages = {1275--1282},
title = {Directed evolution of the 5'-untranslated region of the phoA gene in Escherichia coli simultaneously yields a stronger promoter and a stronger Shine-Dalgarno sequence.},
url = {http://dx.doi.org/10.1002/biot.200600091},
volume = {1},
year = {2006}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Directed evolution has been widely applied for gene improvement through random mutagenesis of coding sequences. Through error-prone PCR both in the coding sequence and the regulatory sequence of E. coli alkaline phosphatase, the cellular enzyme activity has been efficiently enhanced. Sequence analysis revealed that the resultant mutant 34-B12, which showed a sevenfold increased enzyme activity at the cellular level, contains three mutations in the regulatory sequence and another three mutations in the coding sequence. Activity assays of the enzyme containing the corresponding amino acid substitutions proved that the amino acid mutations contribute only to a small portion to the increased cellular enzyme activity. So the mutations in the 5'-untranslated region were analyzed separately and combinationally. The results suggested that one mutation yielded a stronger promoter and the other two mutations both elevated the E. coli alkaline phosphatase expression at the translational level; moreover, a stronger Shine-Dalgarno sequence was generated.
AU  - Huang,X
AU  - Zhang,X-E
AU  - Zhou,Y-F
AU  - Zhang,Z-P
AU  - Cass,AEG
DO  - 10.1002/biot.200600091
EP  - 1282
PY  - 2006///
SP  - 1275
TI  - Directed evolution of the 5'-untranslated region of the phoA gene in Escherichia coli simultaneously yields a stronger promoter and a stronger Shine-Dalgarno sequence.
T2  - Biotechnol J
UR  - http://dx.doi.org/10.1002/biot.200600091
UR  - https://www.ncbi.nlm.nih.gov/pubmed/17109483
VL  - 1
ER  -
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