CN1269948C - Process for changing intact phycobilisome of spirulina platensis - Google Patents
Process for changing intact phycobilisome of spirulina platensis Download PDFInfo
- Publication number
- CN1269948C CN1269948C CN 200510042027 CN200510042027A CN1269948C CN 1269948 C CN1269948 C CN 1269948C CN 200510042027 CN200510042027 CN 200510042027 CN 200510042027 A CN200510042027 A CN 200510042027A CN 1269948 C CN1269948 C CN 1269948C
- Authority
- CN
- China
- Prior art keywords
- phycobilisome
- spirulina plalensis
- solution
- repacking
- spirulina
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to a method for improving the intact phycobilisome of spirulina platensis, which belongs to the technical field of marine biotechnology. The present invention is characterized in that the cyanophyceae spirulina platensis which can be cultivated in a large scale is used as material; firstly, cells are broken by ultrasonic waves, the phycobilisome is dissociated from a thylakoid membrane by a detergent TritonX-100, and then, the TritonX-100, chlorophyll and cell debris are conventionally removed centrifugally to obtain the phycobilisome of spirulina platensis; subsequently, the phycobilisome of spirulina platensis is improved by RPE by a glutaraldehyde crosslinking technology; finally, the pure phycobilisome of spirulina platensis improved by RPE can be obtained through polyethylene glycol precipitation and SepharoseCL-4B column chromatography. The improved phycobilisome can greatly improve detecting quality and detecting sensitivity, and has good application prospects in the process of super-sensitive biomedical detection.
Description
(1) technical field
The present invention relates to a kind of method of changing intact phycobilisome of spirulina platensis, belong to the marine biotechnology field.
(2) background technology
For cellular localization, interaction and the dynamic change thereof of studying protein and other, the researchist is badly in need of new technology and novel material and is realized " sign ", " reading " and " inquiry " to protein and other.Fluorescent mark now commonly used because the restriction of luminescent dye molecule fluorescent characteristic (fluorescence spectrum broad, quantum yield low), can not be applicable to the single-minded sign of high-throughout biomacromolecule far away.
Phycobiliprotein is that a class photosynthesis is caught photopigment-protein complex, mainly is present in blue-green algae, red algae, latent algae and the minority dinoflagellate.In photosynthesis, play a part to catch and transmit luminous energy, have intensive fluorescence.In blue-green algae and red algae, form the supramolecular structure phycobilisome by 2-3 kind phycobiliprotein, form transmission ofenergy sequence efficiently.At the mid-80, the scholar of American Studies algae photosynthesis proposition as fluorescent marker, is used for diagnostic reagent with phycobiliprotein.Because its unique advantage, the diagnostic reagent of phycobiliprotein and phycobiliprotein mark enters the world market in the early 1990s.
Compare with fluorescent marker commonly used, phycobiliprotein has following advantage: production process safety, nontoxic, and luminous energy absorbs strong, the fluorescent yield height surpasses 90%, and bias light interference and false positive rate are low, stable in the scope of pH4-11, can make double-colored, three looks and four color markers.So the range of application of this class reagent constantly enlarges.But,, be not applied to popular reagent for clinical diagnosis as yet owing to cost an arm and a leg.The whole imports of China also only limit to the diagnosis of carrying out with cell streaming instrument.
Phycobiliprotein has its special advantages as fluorescent probe, but one phycobiliprotein molecule, because its molecule is less, the pigment group that contains in the molecule is less, therefore, the brightness that produces fluorescence is not enough, makes in high-resolution detection medium sensitivity not high.U.S. Mo Senman (J.P.Morseman) proposes, and applying immobilized intact phycobilisome can be used for multiple biomedical the detection as fluorescent probe.Because contain more pigment group in the phycobilisome, therefore, its brightness is higher, and excitation wavelength is the absorbing wavelength of phycoerythrin (PE) or Phycocyanins, C-(PC), and fluorescent emission is from APC, therefore, excitation wavelength and emission wavelength are avoided the phase mutual interference at a distance of farther.Spirulina plalensis (Spirulina platensis) is the unicellular filamentous cyanobacteria that can carry out large-scale cultivation at present; it is the good material of preparation intact phycobilisome; the phycobilisome of spirulina plalensis is by Phycocyanins, C-(CPC) and Allophyxoxyanin (APC) and be connected albumen on a small quantity and form; the maximum absorption band 620nm that its efficient excitation wavelength is CPC; its emission peak is the fluorescence emission peak 680nm of APC; stoke shift is 80nm; in order further to strengthen stoke shift; better avoid the interference of exciting light, can manually reequip the spirulina plalensis phycobilisome that is easy to prepare to emitting fluorescence.
(3) summary of the invention
The present invention is directed to the deficiencies in the prior art, a kind of method of reequiping the spirulina plalensis intact phycobilisome is provided.Can further strengthen spirulina plalensis intact phycobilisome stoke shift, better avoid the interference of exciting light emitting fluorescence.
The method of repacking spirulina plalensis intact phycobilisome of the present invention, step is as follows:
(1) is raw material with fresh blue-green algae-spirulina plalensis, is dissolved in Sodium phosphate dibasic-potassium dihydrogen phosphate, adopt the ultrasonic disruption cell.
(2) in the frond of fragmentation, add stain remover, phycobilisome is disintegrated down from thylakoid membrane, centrifugal, remove stain remover and broken cell, the crude extract of preparation phycobilisome.
Add-on the present invention of stain remover is not particularly limited, and is by state of the art, suitable so that phycobilisome is dissociated into from thylakoid membrane.
(3) adopt the glutaraldehyde cross-linking technology, with R-phycoerythrin (RPE) repacking spirulina plalensis phycobilisome, specific as follows:
The crude extract of getting the phycobilisome of step (2) is mixed with solution with Sodium phosphate dibasic-potassium phosphate buffer of 0.9mol/L, get spirulina plalensis phycobilisome solution 0.3mL～0.4mL, mix with 0.07mL～0.09mL R-phycoerythrin solution rapidly, R-phycoerythrin solution is prepared with the 0.1mol/L phosphoric acid buffer, and pH 6.8.Mixed solution is slowly dripped 0.1% (W/V) glutaraldehyde solution 0.02mL～0.04mL of Sodium phosphate dibasic-potassium phosphate buffer preparation with 0.75mol/L while vibrating, 19～22 ℃ were reacted 2 hours～2.5 hours, add the glycine solution 0.04mL of 1mol/L～0.06mL termination reaction 30 minutes～40 minutes, and promptly got the spirulina plalensis phycobilisome of reequiping.
Spirulina plalensis phycobilisome to repacking is further purified, and continues following steps:
(4) with the phycobilisome after the polyethylene glycol precipitation repacking, centrifugal, collecting precipitation.
(5) the phycobilisome precipitation with step (4) gained is dissolved in Sodium phosphate dibasic-potassium dihydrogen phosphate again, again this phycobilisome solution is crossed Sepharose CL-4B column chromatography, collect the purplish red solution of wash-out front end, obtain the changing intact phycobilisome of spirulina platensis of purifying.
The changing intact phycobilisome of spirulina platensis that step (5) is obtained purifying is measured spectrum, detects the repacking effect (see figure 3) of phycobilisome.
In the above steps, the part that is not particularly limited all can adopt state of the art.The detailed operation instructions that also can adopt the present invention to provide below.
The broken concrete operations of above-mentioned steps (1) raw material are: fresh spirulina plalensis 0.75～1.0g is dissolved in 15～20mL1molL
-1Sodium phosphate dibasic-potassium phosphate buffer (pH=6.8～7.0) in, ultrasonic disruption, broken power are 12～15W, broken 5～6 times, each 1.5～2min, 2～3min at interval.
The concrete operations of dissociating of above-mentioned steps (2) phycobilisome are: add stain remover tween X-100 (TritonX-100) in the frond solution of fragmentation, concentration of volume percent 1.9～2.1%, under the room temperature, slowly stirred 1 hour～1.5 hours, centrifugal 3-4 time of 13000rpm～15000rpm removes the cell debris of TritonX-100, chlorophyll and the bottom on top layer, collects out the solution at centrifuge tube middle part, be the phycobilisome crude extract, wherein be mixed with a spot of broken phycobilisome.
The used polyoxyethylene glycol of above-mentioned steps (4) is a polyethylene glycol 6000.
Sodium phosphate dibasic-potassium phosphate buffer pH=6.8～7.0 that above-mentioned steps (5) is used, concentration is 1molL
-1, the phycobilisome precipitation is 1: 1.5～2.0 with the weight ratio of Sodium phosphate dibasic-potassium phosphate buffer.
Above-mentioned steps (5) Sepharose CL-4B post is 2.0cm * 30cm, with pH=6.98 concentration 0.8molL
-1～0.9molL
-1Sodium phosphate dibasic-potassium phosphate buffer wash-out, flow velocity are 0.4mLmin
-1～0.5mLmin
-1
The present invention is a material with blue-green algae spirulina plalensis that can large scale culturing, at first using ultrasound ripple smudge cells, stain remover TritonX-100 obtain the spirulina plalensis phycobilisome from dissociate phycobilisome, conventional centrifugal removal TritonX-100, chlorophyll and cell debris of thylakoid membrane, use the glutaraldehyde cross-linking technology then, with RPE repacking spirulina plalensis phycobilisome, after polyethylene glycol precipitation and Sepharose CL-4B post (2.0 * 30cm) chromatographies obtain the pure spirulina plalensis phycobilisome with the RPE repacking.
The excellent results of present method is: the spirulina plalensis that (1) is made up of CPC and APC with the phycobilisome of artificial culture on a large scale is a material, thereby can obtain starting material cheaply in a large number; (2) prepare phycobilisome in a large number, fast based on simple and easy method, thereby the sucrose density tonsure ultracentrifugation method of having avoided the traditional preparation process phycobilisome is to equipment requirements height, complicated operation and the shortcoming that is difficult to prepare in a large number with conventional equipment and reagent; (3) with excitation wavelength be the RPE of 498nm, rely on the glutaraldehyde cross-linking technology, the spirulina plalensis phycobilisome is reequiped, phycobilisome after the repacking, excitation wavelength are the absorption peak 498nm of RPE, and emission wavelength is fluorescence emission peak 660～680nm of APC, stoke shift reaches 180nm, stoke shift before the repacking is 80nm, and stoke shift strengthens 80～100nm, thereby can effectively avoid exciting light to radiative interference.Simultaneously, owing to contain 6 pigment pigment groups on each RPE, therefore, the phycobilisome after the repacking contains more pigment group, and brightness is higher.Therefore, the phycobilisome after the repacking can significantly improve the quality and the detection sensitivity of detection, has good application prospects in overdelicate biomedical the detection.
(4) description of drawings
Fig. 1 is the absorption spectrum of R-phycoerythrin (curve 1) and spirulina plalensis phycobilisome (curve 2).Can see that by curve 1 the R-phycoerythrin has three absorption peaks, lay respectively at 565nm, 539nm, 498nm, this is the charateristic avsorption band of three peak type R-phycoerythrin, curve 2 can see that the maximum absorption band of spirulina plalensis at 613nm, has acromion at the 650nm place.
Fig. 2 is the cross-linking agent (thick line) of R-phycoerythrin and spirulina plalensis phycobilisome and the absorption spectrum of mixture (fine rule).Compare with Fig. 1, the absorption spectrum of R-phycoerythrin and spirulina plalensis phycobilisome mixture (fine rule among the figure) obviously is the stack of R-phycoerythrin and phycobilisome absorption spectrum, wherein the absorption peak of R-phycoerythrin 539nm becomes the absorption acromion in mixture, and mixture at the absorption peak of ultraviolet region still at 278nm.
Fig. 3 is the cross-linking agent (1) of R-phycoerythrin and spirulina plalensis phycobilisome and the room temperature fluorescence emmission spectrum (excitation wavelength 480nm) of mixture (2).Can see, mixture (curve 2) has maximum emission peak at 578nm under the 480nm optical excitation, this peak is the feature emission peak of R-phycoerythrin, does not have the peak to occur in other positions, illustrate after the R-phycoerythrin absorbs energy and disperse middle noenergy transmittance process with the form of autofluorescence.And the emmission spectrum of cross-linking agent (curve 1) is except that occurring at 578nm the emission peak of R-phycoerythrin, another emission peak appears at 660nm, this is the fluorescence emission peak (or emission peak of Allophyxoxyanin) of phycobilisome, and the fluorescence intensity of the emission peak of R-phycoerythrin obviously reduces, illustrate that the R-phycoerythrin has passed to phycobilisome with portion of energy, prove the crosslinked success of R-phycoerythrin and phycobilisome.
Fig. 4 is the cross-linking agent (1) of R-phycoerythrin and spirulina plalensis phycobilisome and the room temperature fluorescence excitation spectrum (emission wavelength 660nm) of mixture (2).Among the figure, the excitation peak of the 660nm of mixture (curve 2) is at 618nm, and this is the peak of phycobilisome, does not have tangible excitation peak in other positions, illustrates that mixture 660nm fluorescence mainly comes from the photoabsorption of phycobilisome self; And the excitation spectrum of cross-linking agent (curve 1) shows the excitation peak except 618nm, the excitation peak that also has 498nm and two R-phycoerythrin of 568nm, the peak value of 568nm even suitable with the peak value of 618nm, the fluorescence of this explanation cross-linking agent 660nm is to absorb luminous energy by the R-phycoerythrin to produce excited state, and with the fluorescence of transmission ofenergy to the phycobilisome generation.By above analysis as can be known, have only R-phycoerythrin and phycobilisome covalent cross-linking together, just exist energy,, then do not have transmission ofenergy to carry out if both do not have covalent cross-linking together from of the transmission of R-phycoerythrin to phycobilisome.
(5) embodiment
Embodiment 1:
(1) fresh spirulina plalensis 1.0g is dissolved in the 20mL 1molL of pH=6.9
-1In Sodium phosphate dibasic-potassium phosphate buffer, adopt the ultrasonic disruption cell, broken power is 15W, broken 5 times, and each 2min, 2min at interval.
(2) add stain remover TritonX-100 in the frond of fragmentation, frond and stain remover volume ratio are 1: 0.2, under the room temperature, slowly stir 1.5 hours, and phycobilisome is disintegrated down from thylakoid membrane.Centrifugal 4 times of 14000rpm removes the cell debris of TritonX-100, chlorophyll and the bottom on top layer, collects out the solution at centrifuge tube middle part, is the phycobilisome crude extract, wherein is mixed with a spot of broken phycobilisome.
(3) crude extract of getting the phycobilisome of step (2) is mixed with solution with Sodium phosphate dibasic-potassium phosphate buffer pH=6.8～7.0 of 0.9mol/L in 1: 1 ratio, get spirulina plalensis phycobilisome solution 0.3mL, mix with 0.07mL R-phycoerythrin solution rapidly, R-phycoerythrin solution is prepared with the 0.1mol/L phosphoric acid buffer, pH 6.8, protein concentration 1mg/ml.Mixed solution is slowly dripped 0.1% (W/V) glutaraldehyde solution 0.02mL of Sodium phosphate dibasic-potassium phosphate buffer preparation with 0.75mol/L while vibrating, 20 ℃ of reaction 2h, add the glycine solution 0.05mL termination reaction 30min of 1mol/L, promptly get the spirulina plalensis phycobilisome of reequiping.
(4) precipitate phycobilisome with polyethylene glycol 6000, collecting precipitation.
(5) will precipitate and be dissolved in pH=6.9 concentration 1molL again
-1Sodium phosphate dibasic-potassium dihydrogen phosphate in, cross the Sepharose CL-4B column chromatography of 2.0 * 30cm then, with pH=6.98 concentration 0.9molL
-1Sodium phosphate dibasic-potassium phosphate buffer wash-out, flow velocity are 0.45mLmin
-1Collect the blue-violet solution of wash-out front end, obtain the whole phycobilisome of spirulina plalensis of the repacking of purifying.
The whole phycobilisome of the spirulina plalensis of repacking excites with the absorption peak 498nm of R-phycoerythrin, and emission wavelength is the fluorescence emission peak 660nm of APC, stoke shift reaches 160nm, and the stoke shift before the repacking is 80nm, and stoke shift has strengthened 80nm, thereby can effectively avoid exciting light to radiative interference, simultaneously, owing to contain 6 pigment pigment groups on each R-phycoerythrin, therefore, phycobilisome after the repacking contains more pigment group, and brightness is higher.Therefore, the phycobilisome after the repacking can significantly improve the quality and the detection sensitivity of detection.
Embodiment 2:
Different is that step (3) is as follows as embodiment 1:
The crude extract of getting the phycobilisome of step (2) is mixed with solution with Sodium phosphate dibasic-potassium phosphate buffer pH=6.8～7.0 of 0.9mol/L in 1: 1 ratio, get spirulina plalensis phycobilisome solution 0.4mL, mix with 0.08mLR-phycoerythrin solution rapidly, R-phycoerythrin solution is prepared with the 0.1mol/L phosphoric acid buffer, pH 6.8, protein concentration 1.5mg/ml.Mixed solution is slowly dripped 0.1% (W/V) glutaraldehyde solution 0.03mL of Sodium phosphate dibasic-potassium phosphate buffer preparation with 0.75mol/L while vibrating, 19 ℃ were reacted 2.5 hours, add the glycine solution 0.055mL termination reaction 35 minutes of 1mol/L, promptly get the spirulina plalensis phycobilisome of reequiping.
Claims (8)
1, a kind of method of reequiping the spirulina plalensis intact phycobilisome, step is as follows:
(1) is raw material with fresh spirulina plalensis, is dissolved in Sodium phosphate dibasic-potassium dihydrogen phosphate, adopt the ultrasonic disruption cell;
(2) in the frond of fragmentation, add stain remover, phycobilisome is disintegrated down from thylakoid membrane, centrifugal, remove stain remover and broken cell, the crude extract of preparation phycobilisome;
(3) adopt the glutaraldehyde cross-linking technology, with R-phycoerythrin repacking spirulina plalensis phycobilisome, specific as follows:
The crude extract of getting the phycobilisome of step (2) is mixed with solution with Sodium phosphate dibasic-potassium phosphate buffer of 0.9mol/L, get spirulina plalensis phycobilisome solution 0.3mL～0.4mL, mix with 0.07mL～0.09mL R-phycoerythrin solution rapidly, R-phycoerythrin solution is prepared with the 0.1mol/L phosphoric acid buffer, pH6.8; Mixed solution is slowly dripped the 0.1%W/V glutaraldehyde solution 0.02mL～0.04mL of Sodium phosphate dibasic-potassium phosphate buffer preparation with 0.75mol/L while vibrating, 19～22 ℃ were reacted 2 hours～2.5 hours, add the glycine solution 0.04mL of 1mol/L～0.06mL termination reaction 30 minutes～40 minutes, and promptly got the spirulina plalensis phycobilisome of reequiping.
2, the method for repacking spirulina plalensis intact phycobilisome as claimed in claim 1 is characterized in that, the spirulina plalensis phycobilisome of reequiping is further purified, and continues following steps:
(4) with the phycobilisome after the polyethylene glycol precipitation repacking, centrifugal, collecting precipitation;
(5) the phycobilisome precipitation with step (4) gained is dissolved in Sodium phosphate dibasic-potassium dihydrogen phosphate again, again this phycobilisome solution is crossed Sepharose CL-4B column chromatography, collect the purplish red solution of wash-out front end, obtain the changing intact phycobilisome of spirulina platensis of purifying.
3, the method for repacking spirulina plalensis intact phycobilisome as claimed in claim 1 is characterized in that, described step (1) raw material spirulina plalensis 0.75～1.0g is dissolved in 15～20mL 1molL
-1Sodium phosphate dibasic-potassium phosphate buffer in.
4, the method for repacking spirulina plalensis intact phycobilisome as claimed in claim 1 is characterized in that, described step (1) ultrasonic disruption power is 12～15W, broken 5～6 times, and each 1.5～2min, 2～3min at interval.
5, the method for repacking spirulina plalensis intact phycobilisome as claimed in claim 1, it is characterized in that, described step (2) stain remover is tween X-100, concentration of volume percent 1.9～2.1%, under the room temperature, slowly stirred 1 hour～1.5 hours, centrifugal 3-4 time of 13000rpm～15000rpm collects out the solution at centrifuge tube middle part.
6, the method for repacking spirulina plalensis intact phycobilisome as claimed in claim 2 is characterized in that, the used polyoxyethylene glycol of described step (4) is a polyethylene glycol 6000.
7, the method for repacking spirulina plalensis intact phycobilisome as claimed in claim 2 is characterized in that, Sodium phosphate dibasic-potassium phosphate buffer pH=6.8～7.0 that described step (5) is used, and concentration is 1mo1L
-1
8, the method for repacking spirulina plalensis intact phycobilisome as claimed in claim 2 is characterized in that, described step (5) Sepharose CL-4B post is 2.0cm * 30cm, with pH=6.98 concentration 0.8molL
-1～0.9molL
-1Sodium phosphate dibasic-potassium phosphate buffer wash-out, flow velocity are 0.4mLmin
-1～0.5mLmin
-1
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510042027 CN1269948C (en) | 2005-01-17 | 2005-01-17 | Process for changing intact phycobilisome of spirulina platensis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510042027 CN1269948C (en) | 2005-01-17 | 2005-01-17 | Process for changing intact phycobilisome of spirulina platensis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1654628A CN1654628A (en) | 2005-08-17 |
| CN1269948C true CN1269948C (en) | 2006-08-16 |
Family
ID=34894455
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200510042027 Expired - Fee Related CN1269948C (en) | 2005-01-17 | 2005-01-17 | Process for changing intact phycobilisome of spirulina platensis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1269948C (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108508214A (en) * | 2018-05-31 | 2018-09-07 | 福州大学 | A kind of phycoerythrin fluorescence probe and its method quickly detected for aflatoxin B1 |
-
2005
- 2005-01-17 CN CN 200510042027 patent/CN1269948C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN1654628A (en) | 2005-08-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102964608B (en) | Preparation method of carbon quantum dot containing calcium alginate gel for detecting copper ions | |
| CN102329381B (en) | Method for simultaneously separating high-purity phycocyanin and allophycocyanin and application thereof | |
| CN101240010B (en) | Method for fast separating and purifying C-phycocyanin and isophycocyanin from blue algae | |
| CN104140489A (en) | Amphipathy photoswitch fluorescent polymer nano particles and preparing method thereof | |
| CN111289336A (en) | Fungus fluorescent staining solution and preparation method thereof | |
| CN107384395A (en) | The preparation method of water-soluble nitrogen phosphorus boron codope carbon quantum dot | |
| CN106497561B (en) | Method for preparing graphene quantum dot yellow fluorescent powder or mixture thereof and method for preparing white light LED device | |
| CN110373184A (en) | A kind of synthetic method of high yield red fluorescence carbon quantum dot material | |
| CN115124998B (en) | N, S, B doped Chinese herbal medicine residue carbon point, fluorescent probe and application | |
| CN1269948C (en) | Process for changing intact phycobilisome of spirulina platensis | |
| CN101240011B (en) | Fast large preparation method for high-purity isophycocyanin | |
| CN113214301B (en) | Fluoroboropyrrole derivatives with cell membrane labeling function and used for monomolecular positioning super-resolution imaging and monomolecular tracking and application thereof | |
| CN105801562B (en) | A kind of solid broadband red emission luminous organic material and preparation method thereof | |
| CN1273513C (en) | Alcohol soluble high molecular material in poly-fluorene group containing phosphate group and preparation method | |
| CN111548445B (en) | Optical activity AIE fluorescent material and preparation method of aqueous polymer thereof | |
| CN105424930A (en) | Method for rapidly detecting staphylococcus aureus | |
| CN105001141A (en) | Fluoro phenylindole compound, application of fluoro phenylindole compound as red organic luminescent material, and preparation method of fluoro phenylindole compound | |
| CN1654629A (en) | Process for preparing intact phycobilisome | |
| Thimmaraju et al. | Selenium-containing bodipy dyes as photosensitizers | |
| CN115449368B (en) | Nitrogen-doped double-emission fluorescent carbon dot and preparation method and application thereof | |
| CN1271085C (en) | Rapid separating purification of R-phycoerythrin | |
| CN108714223B (en) | Contrast agent with magnetic resonance and fluorescence dual imaging characteristics and preparation method thereof | |
| CN1654627A (en) | Stabilization technology for intact phycobilisomes of red algae and blue algae | |
| CN109734925B (en) | Polymer fluorescence sensor for detecting hypochlorite based on porphyrin comparability, preparation method and application | |
| CN102115779A (en) | Detection method used for detecting variety and quantity of red tide algae |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |