Quantitative specific detection of Staphylococcus aureus based on recombinant lysostaphin and ATP bioluminescence.
- Author:
Yuyuan LI
;
Zhiqiang MI
;
Xiaoping AN
;
Yusen ZHOU
;
Yigang TONG
- Publication Type:Journal Article
- MeSH:
Adenosine Triphosphate;
chemistry;
Chromatography, Affinity;
Escherichia coli;
Luminescent Measurements;
methods;
Lysostaphin;
chemistry;
Recombinant Proteins;
chemistry;
Staphylococcus aureus;
isolation & purification
- From:
Chinese Journal of Biotechnology
2014;30(8):1283-1290
- CountryChina
- Language:Chinese
-
Abstract:
Quantitative specific detection of Staphylococcus aureus is based on recombinant lysostaphin and ATP bioluminescence. To produce recombinant lysostaphin, the lysostaphin gene was chemically synthesized and inserted it into prokaryotic expression vector pQE30, and the resulting expression plasmid pQE30-Lys was transformed into E. coli M15 for expressing lysostaphin with IPTG induction. The recombinant protein was purified by Ni(2+)-NTA affinity chromatography. Staphylococcus aureus was detected by the recombinant lysostaphin with ATP bioluminescence, and plate count method. The results of the two methods were compared. The recombinant lysostaphin was successfully expressed, and a method of quantitative specific detection of S. aureus has been established, which showed a significant linear correlation with the colony counting. The detection method developed has good perspective to quantify S. aureus.
