Two Approaches to Enhance the Processivity and Salt Tolerance of Staphylococcus aureus DNA Polymerase.
Processivity
Recombinase polymerase amplification
Salt tolerance
Sso7d
Thioredoxin-binding domain
Journal
The protein journal
ISSN: 1875-8355
Titre abrégé: Protein J
Pays: Netherlands
ID NLM: 101212092
Informations de publication
Date de publication:
04 2019
04 2019
Historique:
pubmed:
14
2
2019
medline:
26
11
2019
entrez:
14
2
2019
Statut:
ppublish
Résumé
In this article, two engineering-strategies were carried out to enhance the processivity of the DNA polymerase used in recombinase polymerase amplification (RPA). We demonstrate that covalent linkage of a non-specific, double-stranded DNA binding protein, Sso7d, to the large fragment of Staphylococcus aureus Pol I (Sau) caused a moderate enhancement of processivity and a significant improvement in the salt tolerance of Sau. Meanwhile, we provide evidence suggesting that insertion of the thioredoxin-binding domain from bacteriophage T7 DNA polymerase into the analogous position of the large fragment of Sau dramatically enhanced the processivity and mildly increased the salt tolerance of Sau when a host DNA binding protein, thioredoxin, was annexed. Both of these two strategies did not improve the amplifying performance of Sau in RPA, indicating that optimum processivity is crucial for amplifying efficiency.
Identifiants
pubmed: 30759302
doi: 10.1007/s10930-019-09818-7
pii: 10.1007/s10930-019-09818-7
pmc: PMC6486907
doi:
Substances chimiques
Bacterial Proteins
0
DNA-Binding Proteins
0
Recombinant Proteins
0
Thioredoxins
52500-60-4
bacteriophage T7 induced DNA polymerase
EC 2.7.7.-
DNA-Directed DNA Polymerase
EC 2.7.7.7
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
190-198Références
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