| dc.contributor.author |
Chronopoulou, EG |
en |
| dc.contributor.author |
Labrou, NE |
en |
| dc.date.accessioned |
2014-06-06T06:51:02Z |
|
| dc.date.available |
2014-06-06T06:51:02Z |
|
| dc.date.issued |
2011 |
en |
| dc.identifier.issn |
19343655 |
en |
| dc.identifier.uri |
http://dx.doi.org/10.1002/0471140864.ps2606s63 |
en |
| dc.identifier.uri |
http://62.217.125.90/xmlui/handle/123456789/5284 |
|
| dc.subject |
Combinatorial mutation libraries |
en |
| dc.subject |
Mutagenesis |
en |
| dc.subject |
Protein engineering |
en |
| dc.subject.other |
oligonucleotide |
en |
| dc.subject.other |
plasmid DNA |
en |
| dc.subject.other |
primer DNA |
en |
| dc.subject.other |
reagent |
en |
| dc.subject.other |
restriction endonuclease |
en |
| dc.subject.other |
single stranded DNA |
en |
| dc.subject.other |
amino acid substitution |
en |
| dc.subject.other |
article |
en |
| dc.subject.other |
bacterium culture |
en |
| dc.subject.other |
clinical protocol |
en |
| dc.subject.other |
DNA degradation |
en |
| dc.subject.other |
DNA template |
en |
| dc.subject.other |
Escherichia coli |
en |
| dc.subject.other |
gel electrophoresis |
en |
| dc.subject.other |
mutational analysis |
en |
| dc.subject.other |
polymerase chain reaction |
en |
| dc.subject.other |
priority journal |
en |
| dc.subject.other |
protein engineering |
en |
| dc.subject.other |
protein function |
en |
| dc.subject.other |
protein variant |
en |
| dc.subject.other |
site directed mutagenesis |
en |
| dc.subject.other |
structure activity relation |
en |
| dc.title |
Site-saturation mutagenesis: A powerful tool for structure-based design of combinatorial mutation libraries |
en |
| heal.type |
journalArticle |
en |
| heal.identifier.primary |
10.1002/0471140864.ps2606s63 |
en |
| heal.identifier.secondary |
26.6 |
en |
| heal.publicationDate |
2011 |
en |
| heal.abstract |
This unit describes a method for site-saturation mutagenesis (SSM) using PCR amplification with degenerate synthetic oligonucleotides as primers. SSM allows the substitution of predetermined protein sites against all twenty possible amino acids at once. Therefore, SSM is a powerful approach in protein engineering to characterize structure-function relationships, as well as to create improved protein variants. The procedure accepts double-stranded plasmid isolated from the dam+ E. coli strain. The procedure is simple, fast, efficient, and eliminates time-consuming subcloning and ligation steps. © 2011 by John Wiley & Sons, Inc. |
en |
| heal.journalName |
Current Protocols in Protein Science |
en |
| dc.identifier.issue |
SUPPL.63 |
en |
| dc.identifier.doi |
10.1002/0471140864.ps2606s63 |
en |