Signal-enhanced fluorescent protein probe for detecting mercury ions as well as preparation and application of fluorescent protein probe

• Main Authors: YAN YIYU, HUANG JINMIAO, WEI MIN, WANG DAN
• Format: Patent
• Language: Chinese; English
• Published: 21.04.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PHYSICS; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TESTING; VINEGAR; WINE

The invention discloses a signal-enhanced fluorescent protein probe for detecting mercury ions. The fluorescent protein probe has an amino acid sequence as shown in SEQ ID No. 1. According to the invention, amino acids near an EGFP chromophore are mutated, and EGFP is modified into a biosensor sensitive to Hg < 2 + >, i.e. 203 and 205 amino acids near the EGFP chromophore are mutated into cysteine capable of being specifically bound with Hg < 2 + >, and the obtained fluorescent protein double mutant EGFP T203C S205C is transformed into escherichia coli for expression, so that the Hg < 2 + > is detected. The fluorescent protein probe for detecting the mercury ions in the signal enhancement mode is obtained. When Hg < 2 + > is added into expressed escherichia coli, cysteine residues at the 203 site and the 205 site are coordinated with the Hg < 2 + >, so that an electronic environment around a chromophore is changed, and a fluorescence signal is changed. Therefore, the mercury ion signal enhanced green fluoresc