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Development of an artificial zinc finger – Luciferase fusion protein for the rapid detection of Salmonella typhimurium

Authors :
Zhiyang Dong
Yanfeng Zhang
Li Wang
Hanlin Cai
Qi Zhou
Source :
Biochemical and Biophysical Research Communications. 579:35-39
Publication Year :
2021
Publisher :
Elsevier BV, 2021.

Abstract

A novel artificial Zinc finger – luciferase fusion protein was successfully developed for rapid detection of Salmonella typhimurium, a worldwide-distributed foodborne pathogen. The designed Zinc finger (ZF) protein bound specifically to a 12 bp region of the Salmonella spp invasion gene invA. While the luciferase from Gaussia princeps called Gaussia luciferase (Gluc) was for the first time fused with the artificial ZF domain to improve the detection sensitivity. The fusion protein successfully recognized and bound to the synthesized invA dsDNA with high specificity and sensitivity. The detection limit was as low as 10 fmol of dsNDA. Then, the bacteria PCR products were subsequently used to assess the zinc finger – luciferase fusion protein. The final results indicated that the ZF-Gluc fusion protein system could detect S. typhimurium as low as 1 CFU/mL in 2 h after the PCR. Therefore, this study provided us with a novel artificial zinc finger fusion protein and an efficient method to accomplish the rapid detection of the major foodborne pathogen S. typhimurium. In addition, the specific artificial ZF proteins that bund to particular dsDNA sequences could be easily designed, the ZF-Gluc might has broad application prospects in the field of rapid pathogenic bacteria detection.

Details

ISSN :
0006291X
Volume :
579
Database :
OpenAIRE
Journal :
Biochemical and Biophysical Research Communications
Accession number :
edsair.doi.dedup.....61b423ac59c2c844135cf7e318761ab1
Full Text :
https://doi.org/10.1016/j.bbrc.2021.09.033