Luciferase-Zinc-Finger System for the Rapid Detection of Pathogenic Bacteria

被引:12
作者
Shi, Chu [1 ]
Xu, Qing [2 ]
Ge, Yue [1 ]
Jiang, Ling [3 ]
Huang, He [2 ]
机构
[1] Nanjing Tech Univ, Coll Biotechnol & Pharmaceut Engn, 30 Puzhu Rd, Nanjing 211800, Jiangsu, Peoples R China
[2] Nanjing Tech Univ, Sch Pharmaceut Sci, State Key Lab Mat Oriented Chem Engn, 30 Puzhu Rd, Nanjing 211800, Jiangsu, Peoples R China
[3] Nanjing Tech Univ, Coll Food Sci & Light Ind, 30 Puzhu Rd, Nanjing 211800, Jiangsu, Peoples R China
关键词
luciferase; zinc-finger protein; pathogenic bacteria; detection; FIREFLY LUCIFERASE; PCR PRODUCTS; PROTEIN; CLASSIFICATION; DIVERSITY; LINKERS;
D O I
10.1021/acs.jafc.7b02204
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
Rapid and reliable detection of pathogenic bacteria is crucial for food safety control. Here, we present a novel luciferase-zinc finger system for the detection of pathogens that offers rapid and specific profiling. The system, which uses a zinc-finger protein domain to probe zinc finger recognition sites, was designed to bind the amplified conserved regions of 16S rDNA, and the obtained products were detected using a modified luciferase. The luciferase-zinc finger system not only maintained luciferase activity but also allowed the specific detection of different bacterial species, with a sensitivity as low as 10 copies and a linear range from 10 to 10(4) copies per microliter of the specific PCR product. Moreover, the system is robust and rapid, enabling the simultaneous detection of 6 species of bacteria in artificially contaminated samples with excellent accuracy. Thus, we envision that our luciferase-zinc finger system will have far-reaching applications.
引用
收藏
页码:6674 / 6681
页数:8
相关论文
共 30 条
[1]   Rapid and Sensitive Detection of Foodborne Pathogenic Bacteria (Staphylococcus aureus) Using an Electrochemical DNA Genomic Biosensor and Its Application in Fresh Beef [J].
Abdalhai, Mandour H. ;
Fernandes, Antonio Maximiano ;
Bashari, Mohand ;
Ji, Jian ;
He, Qian ;
Sun, Xiulan .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2014, 62 (52) :12659-12667
[2]   Detection of Pathogenic Bacteria by Using Zinc Finger Protein Fused with Firefly Luciferase [J].
Abe, Koichi ;
Kumagai, Takenori ;
Takahashi, Chiaki ;
Kezuka, Aki ;
Murakami, Yoshiyuki ;
Osawa, Yuko ;
Motoki, Hiroaki ;
Matsuo, Takafumi ;
Horiuchi, Michio ;
Sode, Koji ;
Igimi, Shizunobu ;
Ikebukuro, Kazunori .
ANALYTICAL CHEMISTRY, 2012, 84 (18) :8028-8032
[3]  
Abe T. K. K, 2015, J MICROB BIOCH TECHN, V7, P278
[4]  
[Anonymous], EST GLOB BURD FOODB
[5]   Fusion protein linkers: Property, design and functionality [J].
Chen, Xiaoying ;
Zaro, Jennica L. ;
Shen, Wei-Chiang .
ADVANCED DRUG DELIVERY REVIEWS, 2013, 65 (10) :1357-1369
[6]  
Chhabra P, 2016, FOOD MICROBIOL FOOD, P303, DOI 10.1007/978-3-319-30723-7_11
[7]  
Chung HJ, 2013, NAT NANOTECHNOL, V8, P369, DOI [10.1038/NNANO.2013.70, 10.1038/nnano.2013.70]
[8]  
DeLuca M., 1978, METHODS ENZYMOLOGY, V57, P3, DOI DOI 10.1016/0076-6879(78)57003-1
[9]   An analysis of protein domain linkers: their classification and role in protein folding [J].
George, RA ;
Heringa, J .
PROTEIN ENGINEERING, 2002, 15 (11) :871-879
[10]   Thermostabilization of firefly luciferase by in vivo directed evolution [J].
Koksharov, Mikhail I. ;
Ugarova, Natalia N. .
PROTEIN ENGINEERING DESIGN & SELECTION, 2011, 24 (11) :835-844