CN102147364B - Efficient microbial fluorescent sensor for detecting steroids hormone and polycyclic aromatic hydrocarbon - Google Patents
Efficient microbial fluorescent sensor for detecting steroids hormone and polycyclic aromatic hydrocarbon Download PDFInfo
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- CN102147364B CN102147364B CN 201010595097 CN201010595097A CN102147364B CN 102147364 B CN102147364 B CN 102147364B CN 201010595097 CN201010595097 CN 201010595097 CN 201010595097 A CN201010595097 A CN 201010595097A CN 102147364 B CN102147364 B CN 102147364B
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Abstract
The invention relates to an efficient microbial fluorescent sensor for detecting steroids hormone and polycyclic aromatic hydrocarbon, and belongs to the technical fields of molecular biology and microbiology. The efficient microbial fluorescent sensor is characterized in that plasmids pK-3alpha-green fluorescent protein (GFP) 3 and p6 are co-transformed into Escherichia coli; base pairs 468 in a regulatory sequence and base pairs 722 in a GFP reporter gene of a 3alpha-hydroxysteroid dehydrogenase (HSD) promoter and the like are sequentially inserted into the downstream of a cis Beta-galactosidase promoter gene of a recombinant plasmid pK-3alpha-GFP3; and a protein for expressing a regulatory effect in the regulatory sequence of the 3alpha-HSD on the plasmid p6 can be induced when substrates such as estrogen and the like exit in the environment. The invention has the advantages that: the detection range is wider than the other detection methods by over three orders of magnitude; compared with an original GFP-labeled sensor, the efficient microbial fluorescent sensor has wider detection linear range of standard detection liquid of different steroids hormone and polycyclic aromatic hydrocarbon by one to six orders of magnitude because the expression of the plasmids pTopo-3alpha-GFP3 on GFP has amplification and enhancement effects; and the efficient microbial fluorescent sensor has the characteristics of shorter detection time, small detection size, stability, high speed and efficiency.
Description
Technical field
The invention belongs to molecular biology and microbiological technique field.
Background technology
Be accompanied by the fast development of worker, farming, animal husbandry, environment that steroid hormone and multiring aromatic hydrocarbon substance cause and food pollution are serious day by day, have brought significant damage for human lives's health.How to detect the concern of extremely domestic and international government department of this two pollutant and correlative study mechanism fast and efficiently.At present; Steroid hormone and polycyclic aromatic hydrocarbon pollutant detection method are mainly chemistry and biological two big classes in environment, the food: mostly the former is chromatogram and mass spectroscopy, the large-scale costliness of instrument and equipment, and test sample is single; Sample pre-treatments is complicated, and can't detect types of unknown pollutants; Some biological detection method such as cell increment experiment etc., loaded down with trivial details, the consuming time length of process, cost are high, and test sample is single, and is strict to testing environment, experimenter.
Summary of the invention
The objective of the invention is: a kind of high-effective microorganism fluorescent optical sensor that detects steroid hormone and palycyclic aromatic is provided; Plasmid pK-3 α-GFP3 and p6 cotransformation are advanced in the Escherichia coli; Set up a kind of green fluorescent protein microorganisms marked fluorescence detecting system, detected object is steroid hormone and two big types of materials of palycyclic aromatic.
Technical scheme of the present invention is:
The present invention has made up based on Comamonas testosteroni (C.testosteroni, steroid hormone C.T) and palycyclic aromatic microorganism fluorescence detecting system.Comamonas testosteroni can be with steroid hormones such as stosterones as the sole carbon source and the energy, the materials such as palycyclic aromatic of the non-steroid of can also degrading, and (the 3 α-HSD) metabolism of enzyme such as grade digest this type substrate through 3 α-hydroxysteroid dehydrogenase.Green fluorescent protein (GFP) is the reporter gene that is widely used in the biological marker field in recent years; The protein that its gene produced; Light at blue wavelength region excites down; Green fluorescent can be sent, the relative content of specific substrate can be calculated according to its fluorescence intensity through the reference standard curve.
The present invention advances plasmid pK-3 α-GFP3 and p6 cotransformation in the Escherichia coli, has set up a kind of green fluorescent protein microorganisms marked fluorescence detecting system.3 α-regulating and controlling sequence (seeing sequence 2) 470 base-pairs and GFP reporter gene (seeing sequence 3) 717 base-pairs such as HSD promoter have been inserted in the beta galactosidase of taking advantage of a situation (LacZ) promoter gene (seeing sequence 1) downstream at recombinant plasmid pK-3 α-GFP3 successively; Recombinant plasmid p6 contains the whole controlling genes of 3 α-HSD (seeing sequence 4), totally 5257 base-pairs.When having substrates such as estrogen in the environment; Can induce the albumen of the regulating and controlling sequence expression regulation effect of 3 α on the p6 plasmid-HSD; This albumen can make the last promoter of plasmid pK-3 α-GFP3 start and then give expression to GFP albumen, and this moment, the expression strictness of GFP received the control of inducing amount of substrate.
The LacZ promoter gene of taking advantage of a situation among plasmid pK-3 α-GFP3 can act on regulating and controlling sequence and the promoter gene of the 3 α-HSD in itself plasmid, promotes green fluorescent protein great expression thereafter, and fluorescence signal is had amplification, humidification.According to the fluorescence intensity size of this fluorescent optical sensor, can calculate the relative content of specific substrate through the reference standard curve.
Construction method of the present invention is:
1. plasmid pK-3 α-GFP3 and p6 corotation are dissolved Escherichia coli, obtain a strain recombination bacillus coli BL21-GFP through two resistance screenings,
(1) get the competent cell of prepared fresh, cell is evenly suspended to be placed on ice.
(2) add plasmid pK-3 α-GFP3 and p6, ice bath.
(3) 42 ℃ of water-bath heat shocks 90 seconds were placed rapidly 3 minutes on ice.
(4) add the nutrient culture media of 37 ℃ of preheatings, in 37 ℃ of constant-temperature shaking shaking tables, 120 revolution per seconds are cultivated more than 60 minutes.
(5) bacterium is coated on the LB agar plate that contains kanamycins (kannmycin) and the two resistances of ampicillin (ampicillin).
(6) plate is placed more than 1 hour at 37 ℃ of following forwards, after Liquid Absorption to be coated is advanced agar, plate is inverted, and cultivates more than 12 hours.
(7) picking list bacterium colony in liquid LB medium culture, extracts plasmid and identifies that screening positive clone promptly obtains recombination bacillus coli BL21-GFP.
2, set up bioluminescence standard detection curve
(1) will detect the substrate standard items and use anhydrous alcohol solution, compound concentration is 10 respectively
-6M, 10
-9M, 10
-10M, 10
-11M/L, 10
-2The standard detection liquid of M/L.
(2) get bacterium liquid and add standard detection liquid respectively, blank adds absolute ethyl alcohol, leaves standstill, and detects its fluorescence radiation value with fluorescence microplate reader, sets up the standard detection curve.
Detected object of the present invention is:
Two types of materials of steroid hormone and palycyclic aromatic, the sample detection concentration range is: 10
-12To 10
-3Every liter of mole.
The invention has the beneficial effects as follows:
1, the green fluorescent protein microorganisms marked sensor of the present invention's structure carries out the detection of steroid hormone and palycyclic aromatic; Sensing range exceeds more than 3 one magnitude than other detection method; Utilize plasmid pTopo-3 α-GFP3 that the expression of green fluorescent protein is had amplification, enhancement effect, further enlarged the range of linearity (10 that detects
-15To 10
-3Every liter of mole), compare with the sensor of primitive green fluorescin (GFP) mark, the detection range of linearity of the standard detection liquid of different steroid hormones, palycyclic aromatic can exceed 1 to 6 one magnitude.
2, utilize microorganism to make up sensor, detection time is shorter, can accomplish detection with interior in 60 minutes.
3, in the past detection method must be under the prerequisite of known detection material can detection material content, the microorganism fluorescent optical sensor that the present invention makes up can testing environment in the content of unknown steroid hormone, multiring aromatic hydrocarbon substance.
4, can be applicable to the detection of all steroid hormones and palycyclic aromatic in environment, the food.It is little to have detection volume, stable, characteristics fast and efficiently.
Description of drawings
Fig. 1 is plasmid pK-3 α-GFP3 synoptic diagram;
Fig. 2 is a plasmid p6 synoptic diagram;
Fig. 3 is stosterone fluoroscopic examination result;
Fig. 4 is estradiol fluoroscopic examination result;
Fig. 5 is the fluorescent naphthalimide testing result.
Embodiment
The present invention is based on 3 α-hydroxysteroid dehydrogenase (regulating and expressing principle of 3 α-HSD); Utilize recombinant plasmid pK-3 α-GFP3 and p6 corotation to dissolve Escherichia coli; Set up a kind of high-effective microorganism fluorescent optical sensor that detects steroid hormone and palycyclic aromatic; With stosterone, estradiol, naphthalene are that example is set up the standard detection curve respectively.This method has detection time short (within 60 minutes), and sensing range is wide by (10
-12To 10
-6Every liter of mole), highly sensitive (10
-12Level), easy and simple to handle, the obvious advantage of grade with low cost.It is the better method of content of material such as steroid hormone and palycyclic aromatic in testing environment, the food.
Concrete technical scheme is following:
1. plasmid pK-3 α-GFP3 and p6 corotation are dissolved Escherichia coli, obtain a strain recombination bacillus coli BL21-GFP, have following characteristic through two resistance screenings:
(1) comprises recombinant plasmid pK-3 α-GFP3, it is characterized in that: comprise the beta galactosidase of taking advantage of a situation (LacZ) promoter gene, 3alpha-Hydroxysteroid dehydrogenase (the 1190bp base-pair of the regulating and controlling sequence of 3 α-HSD) and reporter gene-green fluorescent protein;
(2) comprise recombinant plasmid p6, it is characterized in that: contain the whole 5.2Kb base-pair of 3 α-HSD complete sequence controlling gene;
(3) can be by steroid hormone and palycyclic aromatic abduction delivering green fluorescent protein:
(4) have kanamycins (kannmycin) and the two resistances of ampicillin (ampicillin).
Above-mentioned recombinant plasmid pK-3 α-GFP3 and p6 make up for this laboratory.
2. recombination bacillus coli BL21-GFP is cultured to OD at 37 ℃
595=1.0 states mix incubation by a certain percentage with bacterium liquid and sample, detect its fluorescent value with fluorescence microplate reader, formulate the fluorescence-content standard curve of different material.Wherein, sample comprises steroid hormone and two types of materials of palycyclic aromatic, and the sample detection scope is: 10
-12To 10
-3Every liter of mole.
Fluoroscopic examination is the result show:
This method is 10 to the stosterone sensing range
-11To 10
-6Every liter of mole, typical curve is y=-52.278x3+657.77x2-1415.8x+40185, R
2=0.9921; The sensing range of estradiol is 10
-12To 10
-3Every liter of mole, standard detection curve: y=-170.92x3+1961.1x2-4755x+39596, R
2=0.9969; The sensing range of naphthalene is 10
-12To 10
-6Standard detection curve: y=-104.92x3+987.54x2-1700.5x+40123, R
2=0.9986.
1, the structure of recombination bacillus coli BL21-GFP
(1) get 100 μ L competent cells of prepared fresh, cell is evenly suspended to be placed on ice.
(2) add plasmid pK-3 α-GFP3 and p6 1.5 a μ L, placed on ice 30 minutes.
(3) 42 ℃ of water-bath heat shocks 90 seconds were placed rapidly 3 minutes on ice.
(4) add the nutrient culture media of 37 ℃ of preheatings of 600 μ L, 37 ℃, 120 revolution per second shaken cultivation 60 minutes;
(5) 200 μ L bacteriums are coated on the LB agar plate that contains kanamycins (kannmycin) and the two resistances of ampicillin (ampicillin).
(6) plate was placed 1 hour at 37 ℃ of following forwards, after Liquid Absorption to be coated is advanced agar, plate was inverted incubated overnight 16 hours.
(7) picking list bacterium colony in liquid LB medium culture, extracts plasmid and identifies that screening positive clone promptly obtains recombination bacillus coli BL21-GFP.
2, set up bioluminescence standard detection curve
(1) the stosterone standard items are used anhydrous alcohol solution, compound concentration is 10 respectively
-6M, 10
-9M, 10
-10M, 10
-11M/L, 10
-2The standard detection liquid of M/L.
(2) the bacterium liquid of getting 100Ml OD595=1.0 adds 2 μ L standard detection liquid respectively, and blank adds 2 μ L absolute ethyl alcohols, 30 ℃ leave standstill 60 minutes after, detect its fluorescence radiation value with fluorescence microplate reader, set up the standard detection curve.
Fluoroscopic examination result such as following table:
Table 1. stosterone fluoroscopic examination result
The stosterone detectability is respectively 10
-11To 10
-6Every liter of mole, fluorescence intensity and concentration meet cubic polynomial curve: y=-52.278x3+657.77x2-1415.8x+40185, R
2=0.9921.
1, the structure of recombination bacillus coli BL21-GFP
(1) get 100 μ L competent cells of prepared fresh, cell is evenly suspended to be placed on ice.
(2) add plasmid pK-3 α-GFP3 and p6 1.5 a μ L, placed on ice 30 minutes.
(3) 42 ℃ of water-bath heat shocks 90 seconds were placed rapidly 3 minutes on ice.
(4) add the nutrient culture media of 37 ℃ of preheatings of 600 μ L, 37 ℃, 120 revolution per second shaken cultivation 60 minutes;
(5) 200 μ L bacteriums are coated on the LB agar plate that contains kanamycins (kannmycin) and the two resistances of ampicillin (ampicillin).
(6) plate was placed 1 hour at 37 ℃ of following forwards, after Liquid Absorption to be coated is advanced agar, plate was inverted incubated overnight 16 hours.
(7) picking list bacterium colony in liquid LB medium culture, extracts plasmid and identifies that screening positive clone promptly obtains recombination bacillus coli BL21-GFP.
2, set up bioluminescence standard detection curve
(1) the estradiol standard items are used anhydrous alcohol solution, respectively compound concentration be 1mM/L, 1 μ M/L, 1nM/L, 1pM/L, standard detection liquid.
(2) get 100 μ L OD595=1.0 bacterium liquid and add 2 μ L standard detection liquid respectively, blank adds 2 μ L absolute ethyl alcohols, 30 ℃ leave standstill 60 minutes after, detect its fluorescence radiation value with fluorescence microplate reader, set up the standard detection curve.
Fluoroscopic examination result such as following table:
Table 2. estradiol fluoroscopic examination result
The estradiol detectability is respectively 10
-12To 10
-3Every liter of mole, fluorescence intensity and concentration meet cubic polynomial curve: y=-170.92x3+1961.1x2-4755x+ 39596, R
2=0.9969.
1, the structure of recombination bacillus coli BL21-GFP
(1) get 100 μ L competent cells of prepared fresh, cell is evenly suspended to be placed on ice.
(2) add plasmid pK-3 α-GFP3 and p6 1.5 a μ L, placed on ice 30 minutes.
(3) 42 ℃ of water-bath heat shocks 90 seconds were placed rapidly 3 minutes on ice.
(4) add the nutrient culture media of 37 ℃ of preheatings of 600 μ L, 37 ℃, 120 revolution per second shaken cultivation 60 minutes;
(5) 200 μ L bacteriums are coated on the LB agar plate that contains kanamycins (kannmycin) and the two resistances of ampicillin (ampicillin).
(6) plate was placed 1 hour at 37 ℃ of following forwards, after Liquid Absorption to be coated is advanced agar, plate was inverted incubated overnight 16 hours.
(7) picking list bacterium colony in liquid LB medium culture, extracts plasmid and identifies that screening positive clone promptly obtains recombination bacillus coli BL21-GFP.
2, set up bioluminescence standard detection curve
(1) the naphthalene standard items are used anhydrous alcohol solution, compound concentration is the standard detection liquid of 1mM/L, 1 μ M/L, 1nM/L, 1pM/L, 1fM/L respectively.
(2) get 100 μ L OD595=1.0 bacterium liquid and add 2 μ L standard detection liquid respectively, blank adds 2 μ L absolute ethyl alcohols, 30 ℃ leave standstill 60 minutes after, detect its fluorescence radiation value with fluorescence microplate reader, set up the standard detection curve.
Fluoroscopic examination result such as following table:
Table 3. fluorescent naphthalimide testing result
The estradiol detectability is respectively 10
-12To 10
-6Every liter of mole, fluorescence intensity and concentration meet cubic polynomial curve: y=-104.92x3+987.54x2-1700.5x+40123, R
2=0.9986.
Claims (5)
1. a high-effective microorganism fluorescent optical sensor that detects steroid hormone and palycyclic aromatic is characterized in that: plasmid pK-3 α-GFP3 and p6 cotransformation are advanced in the Escherichia coli, set up a kind of green fluorescent protein microorganisms marked fluorescence detecting system; 3 α-regulating and controlling sequence 470 base-pairs such as HSD promoter have been inserted in the beta galactosidase promoter gene downstream of taking advantage of a situation at recombinant plasmid pK-3 α-GFP3 successively; Said recombinant plasmid pK-3 α-GFP3 takes advantage of a situation the beta galactosidase promoter gene shown in sequence in the sequence table 1; Said 3 α-regulating and controlling sequence 470 base-pairs such as HSD promoter are shown in sequence in the sequence table 2; And GFP reporter gene 717 base-pairs, sequence 3 is said in said GFP reporter gene 717 base-pairs such as the sequence table; Recombinant plasmid p6 contains the whole controlling genes of 3 α-HSD, totally 5257 base-pairs, and the whole controlling genes of said 3 α-HSD, totally 5257 base-pairs are of sequence in the sequence table 4; When having the estrogen substrate in the environment; Can induce the albumen of the regulating and controlling sequence expression regulation effect of 3 α on the p6 plasmid-HSD; This albumen can make the last promoter of plasmid pK-3 α-GFP3 start and then give expression to GFP albumen, and this moment, the expression strictness of GFP received the control of inducing amount of substrate.
2. a kind of high-effective microorganism fluorescent optical sensor that detects steroid hormone and palycyclic aromatic according to claim 1; It is characterized in that: the LacZ promoter gene of taking advantage of a situation among plasmid pK-3 α-GFP3 can act on regulating and controlling sequence and the promoter gene of the 3 α-HSD in itself plasmid; Promote green fluorescent protein great expression thereafter, fluorescence signal is had amplification, humidification.
3. a kind of high-effective microorganism fluorescent optical sensor that detects steroid hormone and palycyclic aromatic according to claim 1; It is characterized in that:, can calculate the relative content of specific substrate through the reference standard curve according to the fluorescence intensity size of this fluorescent optical sensor.
4. a kind of high-effective microorganism fluorescent optical sensor that detects steroid hormone and palycyclic aromatic according to claim 1 is characterized in that: detected object is steroid hormone and two types of materials of palycyclic aromatic, and the sample detection concentration range is 10
-12To 10
-3Every liter of mole.
5. a kind of high-effective microorganism fluorescent optical sensor that detects steroid hormone and palycyclic aromatic according to claim 1, its construction method is:
A, plasmid pK-3 α-GFP3 and p6 corotation are dissolved Escherichia coli, obtain a strain recombination bacillus coli BL21-GFP through two resistance screenings;
(1) get the competent cell of prepared fresh, cell is evenly suspended to be placed on ice;
(2) add plasmid pK-3 α-GFP3 and p6, ice bath;
(3) 42 ℃ of water-bath heat shocks 90 seconds were placed rapidly 3 minutes on ice;
(4) add the nutrient culture media of 37 ℃ of preheatings, in 37 ℃ of constant-temperature shaking shaking tables, 120 revolution per seconds are cultivated more than 60 minutes;
(5) bacterium is coated on the LB agar plate that contains kanamycins and the two resistances of ampicillin;
(6) plate is placed more than 1 hour at 37 ℃ of following forwards, after Liquid Absorption to be coated is advanced LB agar, plate is inverted, and cultivates more than 12 hours;
(7) picking list bacterium colony in liquid LB medium culture, extracts plasmid and identifies that screening positive clone promptly obtains recombination bacillus coli BL21-GFP;
B, set up bioluminescence standard detection curve
(1) will detect the substrate standard items and use anhydrous alcohol solution, compound concentration is 10 respectively
-6/ Mol/L, 10
-9Mol/L, 10
-10Mol/L, 10
-11Mol/L, 10
-2The standard detection liquid of Mol/L;
(2) get bacterium liquid and add standard detection liquid respectively, blank adds absolute ethyl alcohol, leaves standstill, and detects its fluorescence radiation value with fluorescence microplate reader, sets up the standard detection curve.
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| CN108330139A (en) * | 2017-01-19 | 2018-07-27 | 上海交通大学 | A kind of regulatable putrescine bioluminescence sensor and its application |
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| CN103163114B (en) * | 2013-03-27 | 2015-01-28 | 北京化工大学 | Fluorescent sensor for determining serum protein concentration and manufacturing method thereof |
| CN107505309A (en) * | 2017-07-13 | 2017-12-22 | 长春理工大学 | Detect the preparation method and application of the optical biosensor of steroid hormone in environment |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7135613B1 (en) * | 1999-02-18 | 2006-11-14 | The National University Of Singapore | Chimeric gene constructs for generation of fluorescent transgenic ornamental fish |
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Non-Patent Citations (4)
| Title |
|---|
| 串联质谱同时测定动物组织中22种同化激素.《分析化学》.2009,第37卷(第2期), |
| 于源华 等.生物荧光方法快速测定环境水体中的甾体类激素.《分析化学》.2010,第38卷(第7期), * |
| 陈慧华 等.液相色谱􀀁 |
| 陈慧华 等.液相色谱􀀁串联质谱同时测定动物组织中22种同化激素.《分析化学》.2009,第37卷(第2期), * |
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|---|---|---|---|---|
| CN108330139A (en) * | 2017-01-19 | 2018-07-27 | 上海交通大学 | A kind of regulatable putrescine bioluminescence sensor and its application |
| CN108330139B (en) * | 2017-01-19 | 2021-06-15 | 上海交通大学 | Adjustable putrescine bioluminescence sensor and application thereof |
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