Construction of a RFP-lacZα bicistronic reporter system and its application in lead biosensing

Autor: Lisa Liu, Zhang Wen, Gao Chaoxian, Hui Changye, Hao-qu Zheng, Guo Yan
Jazyk: angličtina
Rok vydání: 2020
Předmět:
0106 biological sciences
Luminescence
Operon
lac operon
Gene Expression
Artificial Gene Amplification and Extension
Biosensing Techniques
01 natural sciences
Biochemistry
Polymerase Chain Reaction
Fluorescence Microscopy
Transcription (biology)
Genes
Reporter
Nucleic Acids
Gene expression
0303 health sciences
Microscopy
Multidisciplinary
Chemistry
Physics
Electromagnetic Radiation
Light Microscopy
Genomics
Cell biology
Lac Operon
Physical Sciences
Medicine
Research Article
Gene prediction
Science
DNA construction
Research and Analysis Methods
Fluorescence
03 medical and health sciences
010608 biotechnology
Escherichia coli
Genetics
RNA
Messenger
Gene Prediction
Operons
Molecular Biology Techniques
Gene
Molecular Biology
030304 developmental biology
Reporter gene
Biology and Life Sciences
Computational Biology
DNA
Vector Cloning
Genome Analysis
Luminescent Proteins
Genes
Lead
Plasmid Construction
mCherry
Cloning
Zdroj: PLoS ONE, Vol 15, Iss 1, p e0228456 (2020)
PLoS ONE
ISSN: 1932-6203
Popis: The combination of a fluorescent reporter and enzymatic reporter provides a flexible and versatile way for the study of diverse biological processes, such as the detection of transcription and translation. Thus, there is an urgent need to develop this novel bifunctional reporter system. This study reports the design, construction, and validation of a new dicistronic mCherry-lacZα reporter system by artificial lac operon and pbr operon models in lacZM15-producing E. coli. It allows two reporter genes to be co-transcribed into a dicistronic mRNA strand, followed by coupled expression of mCherry and lacZα. In artificial lac operons, expression of the downstream lacZα was demonstrated to be positively related to expression of the upstream ORF. In artificial pbr operons, compared with the insertion of downstream full-length lacZ, the insertion of downstream lacZα exerted a slight effect on the response from the upstream mCherry. Furthermore, the downstream lacZα reporter showed stronger response to Pb(II) than the downstream full-length lacZ. Importantly, the response sensitivity of downstream lacZα was still higher than that of upstream mCherry in a dual RFP-lacZα reporter construct. The highly efficient expression profile of the reporter lacZα peptide makes it a preferred downstream reporter in polycistronic constructs. This novel bifunctional reporter system offers a robust tool for biological studies.
Databáze: OpenAIRE
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