CN106706580A - Application of green fluorescent protein mutant as aluminum ion detection probe - Google Patents
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- CN106706580A CN106706580A CN201611065899.0A CN201611065899A CN106706580A CN 106706580 A CN106706580 A CN 106706580A CN 201611065899 A CN201611065899 A CN 201611065899A CN 106706580 A CN106706580 A CN 106706580A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
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Abstract
The invention relates to the application of a green fluorescent protein mutant as an aluminum ion detection probe. A green fluorescent protein serves as a matrix of the green fluorescent protein mutant, and tyrosine in the amino acid sequence is directionally mutated into L-dopa. Such a method is easy to operate and high in selectivity, the aluminum ion detection limit can reach the [mu]mol*L<-1> level, and the detection requirement of a practical sample is met.
Description
Technical field
The present invention relates to aluminium ion detection field, and in particular to a kind of modified enhanced green fluorescent protein is detected as aluminium ion
The application of probe.
Background technology
Aluminium is a kind of common metal, has been widely used in life.But due to food chain accumulation, abuse abuse
Etc. reason, aluminium element easily with aluminium ion ground form influence ecological environment, even into human body, cause cerebral nerve degenerate,
Memory loss etc..Therefore, the Concentration Testing to aluminium ion in different environments has great importance.
At present, with traditional Element detection method, such as atomic absorption method, atomic emissions method and inductivity coupled plasma mass spectrometry
Compared etc. method, fluorescence probe detection has the low advantage of quick, convenient, sensitivity.But, the fluorescence probe of report leads at present
All it is often to be obtained by organic fully synthetic means, preparation process is cumbersome, and a large amount of organic examinations have been used in building-up process
Agent, heavy metal catalyst etc..In order to detect certain metal ion species, secondary pollution is but caused in detection process, against green
Chemical principle.Additionally, in biological medicine analysis, many detections are carried out in live body, it is desirable to which fluorescence probe has high biological
Compatibility, it is nontoxic, be easy to degraded.
Green fluorescent protein (green fluorescent protein, GFP) be it is a kind of it is naturally occurring be green fluorescence
Albumen in jellyfish, is made up of 238 natural amino acids.By after folding, its fluorescence chromophore is wrapped in 11 well
Among beta sheet block, barreled structure is formed.Its fluorescence radiation stabilization, efficiency high is a kind of efficient fluorescence probe.Additionally,
It is not related to the use of any harmful toxic matter due to albumen acquisition process, and its biocompatibility is high, is widely used in point
The research fields such as sub- mark, drug screening, Antibody Fusion, biology sensor.
Ayyadurai et al. (Bioconjugate chemistry, 2011,22 (4):551-555.) in tyrosine defect
Culture has obtained a kind of green fluorescent protein (GFPdopa) of alpha-non-natural amino acid insertion, Argine Monohydrochloride in type Escherichia coli
Tyrosine in sequence is substituted by levodopa.The research has entered research to the property of GFPdopa, but does not disclose it
It is applied to aluminum ions detection.
The content of the invention
The purpose of the present invention is to solve the shortcomings of the prior art, there is provided a kind of modified enhanced green fluorescent protein as aluminium from
The application of sub- detection probe, the method is simple to operate, and aluminium ion test limit is up to μm olL-1Level, and selectivity is high, meets real
The detection of border sample needs.
Technical scheme provided by the present invention is：
A kind of modified enhanced green fluorescent protein as aluminium ion detection probe application, the modified enhanced green fluorescent protein
With green fluorescent protein as parent, tyrosine is directed and sports levodopa in its amino acid sequence.
In above-mentioned technical proposal, because levodopa is in water solution system, there is high compatibility to aluminium ion.Therefore
After the tyrosine in green fluorescent protein chromophore is mutated into levodopa, chromophore can be carried out with free aluminium ion
Coordinate, then produce the change of fluorescence.
The green fluorescent protein parent amino acid sequence is in (Bioconjugate such as Ayyadurai
chemistry,2011,22(4):Disclosed in article 551-555.) delivered.
The amino acid sequence of the green fluorescent protein such as SEQ ID NO:1.
The green fluorescent protein is a kind of tubbiness albumen with fluorescence chromophore, can be entered in normal E. coli
Row expression, between 450~550nm, its ultraviolet maximum absorption band is between 450~550nm for its maximum emission wavelength.
Described modified enhanced green fluorescent protein is expressed in tyrosine defective escherichia coli；The green fluorescence egg
White variant amino acid sum being 238 (not including Histag labels), relative molecular weight 25000~30000 dalton it
Between.
Preferably, the modified enhanced green fluorescent protein by dialysis, it is lyophilized after, be distributed in prepare liquid and examined
Survey.
Preferably, the detection temperature is 4~60 DEG C, the reaction time, prepare liquid pH value was 5~9 in 1~60min.
Further preferably, the detection temperature is 15~40 DEG C, the reaction time in 1~10min, prepare liquid pH value is 6~
8。
Preferably, the prepare liquid is actual contaminated liquid or simulated cushioned liquid.
Preferably, the actual contaminated liquid is by aluminium ion contaminated soil leachate or by the thin of aluminium ion pollution
Born of the same parents' nutrient solution.
Preferably, the simulated cushioned liquid is phosphate buffer (PBS), 2-N- morpholine propane sulfonic acid buffer solutions
(MES), 3-N- morpholines propane sulfonic acid buffer solution (MOPS), 4- HEPESs buffer solution (HEPES) or trihydroxy methyl
Aminomethane-hydrochloride buffer (Tris-HCl).More preferably 3-N- morpholines propane sulfonic acid buffer solution (MOPS) or 4- hydroxyls
Ethyl piperazidine ethanesulfonic acid buffer (HEPES).
Preferably, addition of the modified enhanced green fluorescent protein in prepare liquid is 1~10 μm of olL-1。
Compared with the existing technology, beneficial effects of the present invention are embodied in：
(1) the invention provides a kind of brand-new aluminium ion detection probe, preparation process green non-pollution, and probe is water-soluble
Property high, good biocompatibility, can be used for the detection of biological sample including intracellular aluminium ion concentration；
(2) probe selectivity provided by the present invention is high, in Na+,K+,Ag+,Ca2+,Mg2+,Ba2+,Zn2+,Cd3+,V3+Deng
In the presence of different ions, exist to aluminium ion concentration and obvious response to.
Specific embodiment
Following application examples can make those skilled in the art be more fully understood from the present invention, but do not limit the invention in any way.
The expression of modified enhanced green fluorescent protein
Modified enhanced green fluorescent protein is oriented mutation using the means of the embedded alpha-non-natural amino acid of residue specificity.Will
Tyrosine defective escherichia coli containing green fluorescent protein plasmid DNA enters in minimal medium (Minimal medium)
Row culture, then in the minimal medium containing levodopa, is expressed under IPTG inductions.It is big after expressing completely
Enterobacteria cracks, and cracking supernatant is purified with nickel post, wherein sending the elution fraction as mesh of yellow-green fluorescence
Mark albumen.Freezed again after protein solution is dialysed and obtain greenish yellow solid as modified enhanced green fluorescent protein (GFPdopa).
The detection of aluminium ion concentration is after probe is mixed from different testing samples, by sepectrophotofluorometer meter
Calculate fluorescence intensity change and obtain corresponding aluminium ion concentration.
Application examples 1
1st, 0.14g GFPdopa protein solids are weighed and is dissolved in conduct detection liquid in 1L HEPES cushioning liquid (pH=7.4)
(5μmol·L-1)。
2nd, take appropriate aluminum nitrate and sodium nitrate is dissolved in HEPES cushioning liquid (pH=7.4), be made into 10,20,40,80 μ
mol·L-1Al3+With 50 μm of olL-1Na+The simulating pollution liquid for co-existing in.
3rd, 100 μ L detection liquid (is treated with 100 μ L HEPES buffer solutions (blank) with 100 μ L simulating pollution liquid respectively
Survey liquid) mixing shake up after, stand 5min, control detection temperature be 25 DEG C when, with fluorescence spectrophotometer range meter detection prepare liquid fluorescence
Strength Changes, according to linear equation, are calculated Al3+Concentration.Result shows that the detection probe can detect that simulation is dirty
Aluminium ion concentration in dye liquor, and Na is not received+The interference of presence.
Application examples 2
1st, 0.28g GFPdopa protein solids are weighed and is dissolved in conduct detection liquid in 1L HEPES cushioning liquid (pH=7.0)
(10μmol·L-1)。
2nd, take appropriate aluminum nitrate and potassium nitrate is dissolved in HEPES cushioning liquid (pH=7.0), be made into 10,20,40,80 μ
mol·L-1Al3+With 50 μm of olL-1K+The simulating pollution liquid for co-existing in.
3rd, 200 μ L detection liquid (is treated with 200 μ L HEPES buffer solutions (blank) with 200 μ L simulating pollution liquid respectively
Survey liquid) mixing shake up after, stand 1min, control detection temperature be 25 DEG C when, with fluorescence spectrophotometer range meter detection prepare liquid fluorescence
Strength Changes, according to linear equation, are calculated Al3+Concentration.Result shows that the detection probe can detect that simulation is dirty
Aluminium ion concentration in dye liquor, and K is not received+The interference of presence.
Application examples 3
1st, 0.14g GFPdopa protein solids are weighed and is dissolved in conduct detection liquid in 1L HEPES cushioning liquid (pH=8.0)
(5μmol·L-1)。
2nd, take appropriate aluminum nitrate and silver nitrate is dissolved in HEPES cushioning liquid (pH=8.0), be made into 10,20,40,80 μ
mol·L-1Al3+With 50 μm of olL-1Ag+The simulating pollution liquid for co-existing in.
3rd, it is 50 μ L detection liquid is (to be measured with 50 μ L HEPES buffer solutions (blank) and 50 μ L simulating pollution liquid respectively
Liquid) mixing shake up after, stand 5min, control detection temperature be 15 DEG C when, with fluorescence spectrophotometer range meter detection prepare liquid fluorescence it is strong
Degree change, according to linear equation, is calculated Al3+Concentration.Result shows that the detection probe can detect simulating pollution
Aluminium ion concentration in liquid, and Ag is not received+The interference of presence.
Application examples 4
1st, 0.084g GFPdopa protein solids are weighed and is dissolved in conduct inspection in 1L Tris-HCl cushioning liquid (pH=6.0)
Survey liquid (3 μm of olL-1)。
2nd, take appropriate aluminum nitrate and calcium nitrate is dissolved in Tris-HCl cushioning liquid (pH=6.0), be made into 10,20,40,80 μ
mol·L-1Al3+With 50 μm of olL-1Ca2+The simulating pollution liquid for co-existing in.
3rd, by 100 μ L detection liquid respectively with 100 μ L Tris-HCl buffer solutions (blank) and 100 μ L simulating pollution liquid
(prepare liquid) mixing shake up after, stand 3min, control detection temperature be 15 DEG C when, with fluorescence spectrophotometer range meter detection prepare liquid it is glimmering
Intensity variation, according to linear equation, is calculated Al3+Concentration.Result shows that the detection probe can detect simulation
Aluminium ion concentration in contaminated liquid, and Ca is not received2+The interference of presence.
Application examples 5
1st, 0.14g GFPdopa protein solids are weighed and is dissolved in conduct detection in 1L Tris-HCl cushioning liquid (pH=6.5)
Liquid (5 μm of olL-1)。
2nd, take appropriate aluminum nitrate and barium nitrate is dissolved in Tris-HCl cushioning liquid (pH=6.5), be made into 10,20,40,80 μ
mol·L-1Al3+With 50 μm of olL-1Ba2+The simulating pollution liquid for co-existing in.
3rd, by 200 μ L detection liquid respectively with 200 μ L Tris-HCl buffer solutions (blank) and 200 μ L simulating pollution liquid
(prepare liquid) mixing shake up after, stand 10min, control detection temperature be 25 DEG C when, with fluorescence spectrophotometer range meter detection prepare liquid
Fluorescence intensity change, according to linear equation, is calculated Al3+Concentration.Result shows that the detection probe can detect mould
Intend aluminium ion concentration in contaminated liquid, and do not receive Ba2+The interference of presence.
Application examples 6
1st, 0.28g GFPdopa protein solids are weighed and is dissolved in conduct detection in 1L Tris-HCl cushioning liquid (pH=6.0)
Liquid (10 μm of olL-1)。
2nd, take appropriate aluminum nitrate and magnesium nitrate is dissolved in Tris-HCl cushioning liquid (pH=6.0), be made into 10,20,40,80 μ
mol·L-1Al3+With 50 μm of olL-1Mg2+The simulating pollution liquid for co-existing in.
3rd, by 500 μ L detection liquid respectively with 500 μ L Tris-HCl buffer solutions (blank) and 500 μ L simulating pollution liquid
(prepare liquid) mixing shake up after, stand 10min, control detection temperature be 37 DEG C when, with fluorescence spectrophotometer range meter detection prepare liquid
Fluorescence intensity change, according to linear equation, is calculated Al3+Concentration.Result shows that the detection probe can detect mould
Intend aluminium ion concentration in contaminated liquid, and do not receive Mg2+The interference of presence.
Application examples 7
1st, 0.028g GFPdopa protein solids are weighed and is dissolved in conduct detection liquid in 1L MOPS cushioning liquid (pH=8.0)
(1μmol·L-1)。
2nd, take appropriate aluminum nitrate and zinc nitrate is dissolved in MOPS cushioning liquid (pH=8.0), be made into 10,20,40,80 μm of ol
L-1Al3+With 50 μm of olL-1Zn2+The simulating pollution liquid for co-existing in.
3rd, it is 100 μ L detection liquid is (to be measured with 100 μ L MOPS buffer solutions (blank) and 100 μ L simulating pollution liquid respectively
Liquid) mixing shake up after, stand 10min, control detection temperature be 37 DEG C when, with fluorescence spectrophotometer range meter detection prepare liquid fluorescence it is strong
Degree change, according to linear equation, is calculated Al3+Concentration.Result shows that the detection probe can detect simulating pollution
Aluminium ion concentration in liquid, and Zn is not received2+The interference of presence.
Application examples 8
1st, 0.14g GFPdopa protein solids are weighed and is dissolved in conduct detection liquid (5 in 1L MOPS cushioning liquid (pH=7.0)
μmol·L-1)。
2nd, take appropriate aluminum nitrate and cobalt nitrate is dissolved in MOPS cushioning liquid (pH=7.0), be made into 10,20,40,80 μm of ol
L-1Al3+With 50 μm of olL-1Co2+The simulating pollution liquid for co-existing in.
3rd, by 50 μ L detection liquid respectively with 50 μ L MOPS buffer solutions (blank) and 50 μ L simulating pollutions liquid (prepare liquid)
After mixing shakes up, 3min is stood, when control detection temperature be 37 DEG C, the change of prepare liquid fluorescence intensity is detected with fluorescence spectrophotometer range meter
Change, according to linear equation, be calculated Al3+Concentration.Result shows that the detection probe can be detected in simulating pollution liquid
Aluminium ion concentration, and Co is not received2+The interference of presence.
Application examples 9
1st, 0.28g GFPdopa protein solids are weighed and is dissolved in conduct detection liquid in 1L MOPS cushioning liquid (pH=7.5)
(10μmol·L-1)。
2nd, take appropriate aluminum nitrate and nitric acid vanadium is dissolved in MOPS cushioning liquid (pH=7.5), be made into 10,20,40,80 μm of ol
L-1Al3+With 50 μm of olL-1V3+The simulating pollution liquid for co-existing in.
3rd, by 1000 μ L detection liquid respectively with 1000 μ L MOPS buffer solutions (blank) and 1000 μ L simulating pollution liquid
(prepare liquid) mixing shake up after, stand 10min, control detection temperature be 37 DEG C when, with fluorescence spectrophotometer range meter detection prepare liquid
Fluorescence intensity change, according to linear equation, is calculated Al3+Concentration.Result shows that the detection probe can detect mould
Intend aluminium ion concentration in contaminated liquid, and do not receive V3+The interference of presence.
Application examples 10
1st, 0.14g GFPdopa protein solids are weighed and is dissolved in conduct detection liquid in 1L HEPES cushioning liquid (pH=7.4)
(5μmol·L-1)。
2nd, appropriate soil is taken, 10000rpm centrifuging and taking supernatants after being mixed with MOPS cushioning liquid, and add appropriate nitric acid
Aluminium, is made into 10,20,40,80 μm of olL-1Al3+Actual contaminated liquid.
3rd, 100 μ L detection liquid (is treated with 100 μ L HEPES buffer solutions (blank) with the 100 actual contaminated liquids of μ L respectively
Survey liquid) mixing shake up after, stand 5min, control detection temperature be 25 DEG C when, with fluorescence spectrophotometer range meter detection prepare liquid fluorescence
Strength Changes, according to linear equation, are calculated Al3+Concentration.Result shows that the detection probe can detect that simulation is dirty
Aluminium ion concentration in dye liquor, and do not disturbed by other materials in soil extract.
Application examples 11
1st, weigh during 0.084g GFPdopa protein solids are dissolved in 1L HEPES cushioning liquid (pH=7.4) cushioning liquid and make
It is detection liquid (3 μm of olL-1)。
2nd, appropriate aluminum nitrate is added in the DMEM culture mediums for cultivating cell, 10,20,40,80 μm of olL are made into-1Al3 +Actual contaminated liquid.
3rd, by 2000 μ L detection liquid respectively with 2000 μ L HEPES buffer solutions (blank) and the 2000 actual contaminated liquids of μ L
(prepare liquid) mixing shake up after, stand 3min, control detection temperature be 37 DEG C when, with fluorescence spectrophotometer range meter detection prepare liquid it is glimmering
Intensity variation, according to linear equation, is calculated Al3+Concentration.Result shows that the detection probe can detect simulation
Aluminium ion concentration in contaminated liquid, and do not disturbed by other materials in cell culture fluid.
Application examples 12
1st, after just expressing the Escherichia coli centrifugation for finishing, it is resuspended in HEPES cushioning liquid (pH=7.4), makes OD
(600nm) value is 0.1, used as detection liquid.
2nd, HEPES cushioning liquid (pH=7.4) is dissolved in taking appropriate aluminum nitrate, be made into 10,20,40,80 μm of olL-1Al3 +Simulating pollution liquid.
3rd, by 1000 μ L detection liquid respectively with 1000 μ L HEPES buffer solutions (blank) and the 1000 actual contaminated liquids of μ L
(prepare liquid) mixing shake up after, stand 10min, control detection temperature be 37 DEG C when, with fluorescence spectrophotometer range meter detection prepare liquid
Fluorescence intensity change, according to linear equation, is calculated Al3+Concentration, while the sight of confocal fluorescent microscope can also be used
Examine Escherichia coli change in fluorescence situation.Result shows that the detection probe can be with aluminium ion concentration, Ke Yiyong in detection bacterium body
In internal (in vivo) aluminium ion concentration detection.
SEQUENCE LISTING
<110>Zhejiang University
<120>A kind of modified enhanced green fluorescent protein as aluminium ion detection probe application
<130>
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 238
<212> PRT
<213>Artificial sequence
<400> 1
Met Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val
1 5 10 15
Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Arg Gly Glu
20 25 30
Gly Glu Gly Asp Ala Thr Asn Gly Lys Ile Thr Leu Lys Leu Ile Cys
35 40 45
Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Cys
50 55 60
Gly Tyr Gly Val Gln Cys Phe Ala Arg Tyr Pro Asp His Met Lys Arg
65 70 75 80
His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg
85 90 95
Thr Ile Ser Phe Lys Asp Asp Gly Thr Phe Lys Thr Arg Ala Glu Val
100 105 110
Lys Phe Glu Gly Asp Thr Ile Val Asn Arg Ile Lys Leu Lys Gly Ile
115 120 125
Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn
130 135 140
Phe Asn Ser His Asn Lys Tyr Ile Thr Ala Asp Lys Gln Lys Asn Gly
145 150 155 160
Ile Lys Ala Asn Phe Lys Ile Arg His Asn Val Glu Asp Gly Ser Val
165 170 175
Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro
180 185 190
Arg Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Val Ile Leu
195 200 205
Glu Asp Pro Asn Glu Lys Arg Asp His Met Val Leu His Glu Phe Val
210 215 220
Thr Ala Ala Gly Ile Thr His Gly Met Asp Glu Leu Tyr Lys
225 230 235
Claims (8)
1. a kind of modified enhanced green fluorescent protein as aluminium ion detection probe application, it is characterised in that the green fluorescence
With green fluorescent protein as parent, tyrosine is directed and sports levodopa protein mutant in its amino acid sequence.
2. modified enhanced green fluorescent protein according to claim 1 as aluminium ion detection probe application, its feature exists
In, the modified enhanced green fluorescent protein by dialysis, it is lyophilized after, be distributed in prepare liquid and detected.
3. modified enhanced green fluorescent protein according to claim 2 as aluminium ion detection probe application, its feature exists
In the detection temperature is 4~60 DEG C, and the reaction time, prepare liquid pH value was 5~9 in 1~60min.
4. modified enhanced green fluorescent protein according to claim 3 as aluminium ion detection probe application, its feature exists
In the detection temperature is 15~40 DEG C, and the reaction time, prepare liquid pH value was 6~8 in 1~10min.
5. modified enhanced green fluorescent protein according to claim 2 as aluminium ion detection probe application, its feature exists
In the prepare liquid is actual contaminated liquid or simulated cushioned liquid.
6. modified enhanced green fluorescent protein according to claim 5 as aluminium ion detection probe application, its feature exists
In the actual contaminated liquid is the cell culture fluid polluted by aluminium ion contaminated soil leachate or by aluminium ion.
7. modified enhanced green fluorescent protein according to claim 5 as aluminium ion detection probe application, its feature exists
In the simulated cushioned liquid is phosphate buffer, 2-N- morpholine propane sulfonic acid buffer solution, 3-N- morpholines propane sulfonic acid buffering
Liquid, 4- HEPESs buffer solution or tris-HCI buffer.
8. modified enhanced green fluorescent protein according to claim 2 as aluminium ion detection probe application, its feature exists
In addition of the modified enhanced green fluorescent protein in prepare liquid is 1~10 μm of olL-1。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112179876A (en) * | 2019-07-02 | 2021-01-05 | 南京工业大学 | Method for detecting levodopa and tyrosinase by in-situ formation of fluorescent copolymer |
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| CN102952181A (en) * | 2011-08-26 | 2013-03-06 | 南京大学 | Fluorescence sensor protein, plasmid or cell for detecting cuprous ions, DNA (deoxyribonucleic acid) encoding protein and preparation method |
| CN105585625A (en) * | 2014-10-31 | 2016-05-18 | 北京义翘神州生物技术有限公司 | Enhanced green fluorescence protein |
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| KR20120110633A (en) * | 2011-03-30 | 2012-10-10 | 영남대학교 산학협력단 | Fluorescent protein containing l-3, 4-dihydroxyphenylalanine as a metal biosensor |
| CN102952181A (en) * | 2011-08-26 | 2013-03-06 | 南京大学 | Fluorescence sensor protein, plasmid or cell for detecting cuprous ions, DNA (deoxyribonucleic acid) encoding protein and preparation method |
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| CN112179876A (en) * | 2019-07-02 | 2021-01-05 | 南京工业大学 | Method for detecting levodopa and tyrosinase by in-situ formation of fluorescent copolymer |
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