CN102120968A - Baker's yeast with high sensibility on oxidative damage and preparation method thereof - Google Patents

Baker's yeast with high sensibility on oxidative damage and preparation method thereof Download PDF

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CN102120968A
CN102120968A CN 201010594171 CN201010594171A CN102120968A CN 102120968 A CN102120968 A CN 102120968A CN 201010594171 CN201010594171 CN 201010594171 CN 201010594171 A CN201010594171 A CN 201010594171A CN 102120968 A CN102120968 A CN 102120968A
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yeast
minutes
yap1
bread
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戴和平
肖伟
张超
张敏
张晓华
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Institute of Hydrobiology of CAS
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Abstract

The invention discloses a baker's yeast with high sensibility on oxidative damage and a preparation method thereof, relating to the monitoring of microorganisms modified by a genetic gene to environmental chemistry pollutants. After being modified by a gene technology, the yeast improves the sensitivity to an oxidative damage reagent and ensures wider detection range and higher sensitivity to the environmental pollutants. The yeast is characterized in that an upstream regulatory gene YAP1 of the oxidative damage responding and modifying gene is knocked out on the basis of the original high-permeability baker's yeast, and a yeast mutant strain capable of knocking five genes of yap1, cwp1, cwp2, snq2 and pdr5 with high sensitivity to oxidative damage and super-high permeability on high molecular weight chemicals is obtained. A DNA damage detection element RNR3-1acZ is transferred into the baker's yeast, thus the sensitivity of a detection system to oxidized type genotoxic carcinogens is greatly improved. The invention is more suitable for the detection of chemical carcinogens with low dosage in environment.

Description

A kind of bread yeast to the oxidative damage hypersensitivity and preparation method thereof
Technical field
The present invention relates to biotechnology, relate in particular to through of the monitoring of genetic microorganism of transforming to environmental chemical pollutants.
Background technology
Bread yeast is a kind of simple eukaryote, and unicellular, genetic modification is simple to operate, cultivates easy quick and environmental friendliness, is suitable as a kind of model animals and is applied in the life science.In environmental pollution detection technique field, bread yeast is more and more come into one's own as a kind of simple eukaryote at present, and some are that the detection system of model animals is developed with the bread yeast.
We are on a kind of basis of bread yeast (application number is 200910272505.2) of ultrahigh permeability, yeast oxidative damage response is knocked out with the upstream regulatory gene YAP1 that repairs gene, and having obtained a strain not only can have ultrahigh permeability but also to the extremely sensitive yeast mutant of oxidative damage chemicals to the macromolecule chemicals.YAP1 is the transcriptional regulator that a class plays an important role in response to oxidative stress, is a kind of b-zip (Basic leucine zipper) albumen, is a member in the transcriptional activators AP-1 family, is regulating and control the anti-oxidant expression of gene of many keys.Coerce down at oxyradical, YAP1 albumen can be assembled in nucleus, regulation and control are transcribing of 70 several genes nearly, comprising with a series of albumen of keeping redox environment in the cell, as thioredoxin reductase (TRR), catalase (CTT1), Trx (TRX2), superoxide-dismutase (SOD1), cytochrome c oxidase (CCP1) and a series of and synthetic expression that reaches circulation related gene (GSH1, GTT1 and GPX2) of Triptide.Therefore after YAP1 genetically deficient, can effectively improve the susceptibility of yeast cell to oxidative damage, YAP1 is as important transcriptional regulator simultaneously, play keying action in dna damage is replied, the bread yeast of YAP1 genetically deficient has also shown the susceptibility to many non-oxidizable dna damages.Therefore we determine to utilize the gene knockout technology to knock out the YAP1 gene on ultrahigh permeability zymic basis, set up a kind of bread yeast mutant strain to the oxidative damage hypersensitivity, are applied to the detection of oxidized form genotoxic carcinogens in the environmental pollution.
Gene knockout is a kind of sophisticated Protocols in Molecular Biology that grows up from the end of the eighties, is the technology that makes body special genes inactivation or disappearance by certain approach.Gene knockout on the ordinary meaning mainly is to use dna homology reorganization principle, substitutes target fragment with the homologous fragment that designs, thereby reaches the purpose of gene knockout.Because the full gene sequencing of bread yeast is finished, thus can be according to the sequence of goal gene, and design homologous gene sequence knocks out goal gene.The present invention has analyzed the effect of important transcriptional regulator YAP1 in the yeast, applying gene knocks out technology on the basis of ultrahigh permeability bread yeast (number of patent application is 200910272505.2), the YAP1 gene is knocked out, improved the susceptibility of yeast greatly oxidative damage reagent.
Summary of the invention
The objective of the invention is, a kind of bread yeast to the oxidative damage hypersensitivity is provided, this bread yeast is on the basis of ultrahigh permeability yeast (number of patent application is 200910272505.2), have new bacterial strain, be mainly used in the oxidized form genotoxic carcinogens in the testing environment the oxidative damage hypersensitivity.This yeast mutant not only to the macromolecule chemicals height permeability is arranged, also the oxidative damage effect to chemicals has hypersensitivity, in order to the detection sensitivity of the DNA oxidative damage effect that improves environmental chemical pollutants.Another object of the present invention is that a kind of preparation method to oxidative damage hypersensitivity bread yeast is provided.
In order to achieve the above object, the present invention adopts following technical scheme:
On the basis of ultrahigh permeability bread yeast (number of patent application is 200910272505.2), knock out the YAP1 gene, obtain disappearance Yap1cwp1cwp2pdr5snq2The bread yeast of five genes.
A kind of preparation method to oxidative damage hypersensitivity bread yeast is: extract the bread yeast genomic dna, pass through Auele Specific Primer, with the bread yeast genomic dna is template, pcr amplification obtains the YAP1 gene, structure obtains the yap1 Δ:: hisG-URA3-hisG knocks out assembly, and change this assembly over to ultrahigh permeability bread yeast, and homologous recombination takes place, obtain the bacterial strain of five genetically deficient hypersensitivities by screening.Because the homologous recombination between the hisG sequence, the URA3 gene can be cut, and the URA3 gene can continue property mark alternatively.To carry URA3 gene (uridylic synthase gene) again as selected marker RNR3-LacZThe gene fusion plasmid is transferred to Yap1cwp1cwp2pdr5snq2In the five gene knockout bread yeast mutant strains, obtain can be used for detecting the hypersensitivity bread yeast of oxidized form genotoxic carcinogens.
Genetoxic measuring element used in the present invention is RNR3-lacZThis detection system is model animals with the bread yeast, promotor and reporter gene lacZ(beta-galactosidase enzymes with bread yeast RNR3 gene) syzygy as measuring element, make the RNR3 gene expression dose can be by measuring that the zymetology color reaction be monitored and quantitatively.RNR3 is the big subunit of bread yeast ribonucleotide reductase, and ribonucleotide reductase then is the rate-limiting enzyme that the synthetic required deoxynucleotide of catalytic dna produces.The mRNA transcriptional level of RNR3 is lower under usual conditions, and higher abduction delivering level is but arranged.Its abduction delivering is only relevant with dna damage, and irrelevant with common cell growth cycle.Many environment genotoxic carcinogenses bring out cancer by causing dna damage, so this detection system is developed into a kind of novel detection system of environmental chemical carcinogen.The present invention is by measuring RNR3-lacZThe abduction delivering level judge the raising of yeast cell to the sensitivity of oxidized form genotoxic carcinogens.
Advantage of the present invention and effect:
Yeast of the present invention is compared with wild-type yeast and ultrahigh permeability bread yeast, and the susceptibility of oxidized form genotoxic carcinogens is obviously improved.Wild-type yeast and high-permeability yeast all can not damage by detected tertbutyl peroxide (t-BHP), and Yap1cwp1cwp2pdr5snq2Five gene knockout bread yeasts can detect tertbutyl peroxide (t-BHP) damage; Yap1cwp1cwp2pdr5snq2Five gene knockout bread yeasts have improved 3 times for the detection sensitivity of phleomycin (Phleomycin) than wild-type yeast, have improved 2 times than high-permeability yeast.Therefore Yap1cwp1cwp2pdr5snq2Five gene knockout bread yeasts have improved RNR3-lacZThe detection sensitivity of system makes it more help monitoring to the genotoxic carcinogens that exists with lower concentration in the environment.
Description of drawings
Fig. 1, different bread yeast transgenation strains RNR3-lacZDetection system is to the comparison of DNA oxidative damage reagent tertbutyl peroxide (t-BHP) detection sensitivity.
△: Yap1cwp1cwp2pdr5snq2Five gene knockout yeast; ▼: ultrahigh permeability bread yeast; Zero: Yap1Single-gene knocks out yeast; ●: wild-type yeast.
Fig. 2, different bread yeast transgenation strains RNR3-lacZDetection system is to the comparison of macromolecule DNA oxidative damage reagent phleomycin (Phleomycin) detection sensitivity.
△: Yap1cwp1cwp2pdr5snq2Five gene knockout yeast; ▼: ultrahigh permeability bread yeast; Zero: Yap1Single-gene knocks out yeast; ●: wild-type yeast.
Embodiment
Prepare a kind of method and comprise the following step oxidative damage hypersensitivity bread yeast:
(1), from wild bread yeast, extract genomic dna:
1a, get bread yeast, be inoculated in the YPD nutrient solution, at 30 ℃, under 200 rev/mins of conditions, shaking culture 16 hours obtains bread microzyme liquid;
Wherein: the formulation weight per-cent of YPD nutrient solution is: yeast extract paste 1%, and peptone 2%, glucose 2% the rest is water;
1b, get 1.5 milliliters of bread microzyme liquid, with 4000 rev/mins centrifugal 2 minutes, throw out is suspended from 200 microlitres and extracts in the damping fluid;
Wherein: the prescription that extracts damping fluid is: triton x-100 2%, and sodium laurylsulfonate 1%, 0.1 mole in sodium-chlor, ethylenediamine tetraacetic acid (EDTA) 1 mmole, Tri(Hydroxymethyl) Amino Methane Hydrochloride 10 mmoles the rest is water, and pH 8.0;
1c, add 0.3 gram pickling glass pearl, 100 microlitre phenol, 100 microlitre chloroforms, shake after 3 minutes, with 12000 rev/mins centrifugal 10 minutes, the absorption supernatant liquid;
1d, add the dehydrated alcohol of 2 times of the liquid volumes of getting, left standstill 30 minutes at-20 ℃, with 12000 rev/mins centrifugal 15 minutes, remove supernatant, taking precipitate;
1e, throw out is dissolved in 20 microlitre sterilized waters, is pastoris genomic dna solution;
(2), make up the yap1 Δ:: hisG-URA3-hisG gene knockout assembly:
2a, at first utilize round pcr, by upstream primer YAP1-1:GGA TGC GTA AGG TCT TAA GAG and downstream primer YAP1-2:CAG TGG TTC TGC AGG TAC GG, the pastoris genomic dna fragment amplification of one section 2.5Kb is come out, wherein include open reading frame and the upstream homologous fragment of 0.2Kb and the downstream homologous fragment of 0.35Kb of YAP1.
Wherein: the condition of PCR is: 94 ℃ 5 minutes; 94 ℃ 45 seconds, 52 ℃ 1 minute, 72 ℃ 1 minute, 30 circulations; 72 ℃ 10 minutes;
2b, the pcr amplified fragment of the above-mentioned YAP1 of containing gene is cloned on the pGEM-T carrier (Japanese Takara company), uses BamHIRestriction endonuclease and BstEIIRestriction endonuclease downcuts the segment of one section 1.8kb, contains the joint of BamHI restriction enzyme site in the replacement;
2c, usefulness BamHIRestriction endonuclease and The BglII restriction endonucleaseFrom pNKY51 carrier (reference: Selectable cassettes for simplified construction of yeast gene disruption vectors. Earley, Marie C.and Crouse, Gray F.Gene. 1996,196 (1): 111-113.) go up one section 3.8-kb segment that contains hisG-URA3-hisG of cutting-out, insert the BamHI site in the 2b joint;
2d, usefulness AflII and EcoRIEndonuclease digestion obtains the yap1 Δ:: hisG-URA3-hisG gene knockout assembly;
(3), with the yap1 Δ:: hisG-URA3-hisG gene knockout assembly changes ultrahigh permeability bread yeast over to, and screening obtains yap1cwp1cwp2pdr5snq2 five gene knockout bread yeasts:
3a, ultrahigh permeability bread yeast is inoculated into 2 milliliters of YPD nutrient solutions, 30 ℃, under 200 rev/mins of oscillating conditions, cultivated 16 hours;
3b, add 3 milliliters of YPD nutrient solutions again, 30 ℃, 200 rev/mins, cultivated 4 hours, obtain ultrahigh permeability bread yeast bacterium liquid;
3c, get 1.5 milliliters of ultrahigh permeability bread yeast bacterium liquid, with 2500 rev/mins centrifugal 2 minutes, taking precipitate;
3d, with throw out be suspended from 100 the milli rub/liter 400 microlitre Lithium Acetates in, with 2500 rev/mins centrifugal 2 minutes, taking precipitate;
3e, with throw out be suspended from 100 the milli rub/liter 100 microlitre Lithium Acetates in;
3f, smart single stranded DNA 2 microlitres of the salmon of 2.0 mg/ml were boiled 5 minutes, behind the ice bath with the yap1 Δ:: hisG-URA3-hisG gene knockout assembly 0.1～1 microgram joins in the Lithium Acetate of step 3e, and room temperature was placed 5 minutes;
3g, to add 280 microlitre concentration again be 50% Macrogol 4000;
3h, vibrate to mixing, 30 ℃ left standstill 30 minutes;
3i, add 39 microlitre dimethyl sulfoxide (DMSO), 42 ℃ of heat shocks 5 minutes;
Centrifugal 2 minutes of 3j, 4000 rev/mins, taking precipitate, and throw out is suspended from the 200 microlitre sterilized waters;
Centrifugal 2 minutes of 3k, 4000 rev/mins, taking precipitate is suspended from the 100 microlitre sterilized waters, be suspended from again in the non-selective substratum, grow overnight, and then be applied to 5-fluororotic acid selective medium flat board, cultivated 3 days in 30 ℃ of incubators, grow single bacterium colony, be Yap1cwp1cwp2pdr5snq2Five gene knockout bread yeasts, just of the present invention a kind of to oxidative damage hypersensitivity bread yeast;
Wherein the preparation method of 5-fluororotic acid selective medium flat board is: yeast extract paste 1%, and peptone 2%, glucose 2%, agar powder 2% the rest is water and placed 121 ℃ of following autoclavings 20 minutes, when treating that temperature drops to 50 ℃, adds 5-fluororotic acid 0.1%.
Wherein the preparation method of non-selective substratum is: yeast extract paste 1%, and peptone 2%, glucose 2%, agar powder 2% the rest is water and placed 121 ℃ of following autoclavings 20 minutes.
A kind of oxidative damage hypersensitivity bread yeast is made up with of the present invention RNR3-lacZDetection system is used to detect the oxidized form genotoxic carcinogens, and its method follows these steps to carry out:
1, owing to the homologous recombination between the hisG sequence, Yap1cwp1cwp2pdr5snq2Can be cut fall as the URA3 gene (uridylic synthase gene) of selective marker in the five gene knockout bread yeasts, URA3 can be used for the selected marker of next step, will carry URA3 gene (uridylic synthase gene) as selected marker RNR3-LacZGene fusion plasmid pZZ2(reference: Isolation of crt mutants constitutive for transcription of the DNA damage inducible gene RNR3 in Saccharomyces cerevisiae. Zhou Z, Genetics. 1992 Aug; 131 (4): 851-66.) change over to by the Lithium Acetate method for transformation Yap1cwp1cwp2pdr5snq2In the five gene knockout bread microzymes, and, promptly obtain hypersensitivity yeast detection bacterial strain to the oxidized form genotoxic carcinogens by SD-Ura selectivity plate screening positive strain.
Wherein: the prescription of SD-Ura selectivity flat board is: yeast nitrogen does not have amino acid and does not have ammonium sulfate 0.17 %, ammonium sulfate 0.5%, and glucose 2%, VITAMIN B4 0.2%, tryptophane 1%, Histidine 1%, leucine 1%, Methionin 1%, agar powder 2% the rest is water;
2, with of the present invention a kind of to oxidative damage hypersensitivity yeast detection inoculation in SD-Ura selectivity nutrient solution, 30 ℃, 200 rev/mins of following shaking culture 16 hours obtain a kind of to oxidative damage hypersensitivity bread yeast liquid;
3,0.5 milliliter of yeast liquid is inoculated in 2.5 milliliters of SD-Ura selectivity nutrient solutions, 30 ℃, 200 rev/mins of following shaking culture are to OD 600Value between 0.15～0.20;
Wherein: the prescription of SD-Ura selectivity nutrient solution is: yeast nitrogen does not have amino acid and does not have ammonium sulfate 0.17 %, ammonium sulfate 0.5%, and glucose 2%, VITAMIN B4 0.2%, tryptophane 1%, Histidine 1%, leucine 1%, Methionin 1% the rest is water;
4, in SD-Ura selectivity nutrient solution, add oxidative dna damage reagent tertbutyl peroxide, cultivated 4 hours;
5, with spectrophotometric determination density OD 600Value;
6, get 2 ml yeast bacterium liquid, 2500 rev/mins centrifugal 2 minutes, taking precipitate;
7, throw out is suspended from 1 milliliter of Z damping fluid, adds 0.1% sodium laurylsulfonate, 50 microlitres, 50 microlitre chloroforms, high speed vortex 10 seconds breaks yeast cell;
Wherein: the prescription of Z damping fluid is: Na 2HPO 460 mmoles, NaH 2PO 440 mmoles, KCl 10 mmoles, MgSO 41 mmole, 2 mercapto ethanol 40 mmoles the rest is water; PH 7.0;
8, O-nitrophenyl-200 microlitres that add 4 mg/ml are incubated 20 minutes down at 30 ℃;
9, add 1 mole of Na again 2CO 3500 microlitre termination reactions;
10,3500 rev/mins centrifugal 10 minutes, get supernatant, measure OD 420Absorption value;
11, the OD that records according to step 5 600The OD that value and step 10 record 420Absorption value is calculated beta-galactosidase enzymes vigor: S with following formula A-gal=(1000 * OD 420)/(reaction times * bacteria liquid amasss * OD 600), wherein: S A-galExpression beta-galactosidase enzymes vigor;
12, according to the bread microzyme liquid beta-galactosidase enzymes vigor of above method replicate(determination) without the dna damage agent treated, in contrast, when positive greater than 2 the time through the ratio of the bread microzyme liquid beta-galactosidase enzymes energy value of dna damage agent treated and the bread microzyme liquid beta-galactosidase enzymes energy value of contrast, show to detect in the thing and contain genotoxic carcinogens, in detecting thing, contain under the identical situation of genotoxic carcinogens ratio, ratio is big more, shows bread yeast RNR3-lacZThe detection sensitivity of system is high more.

Claims (2)

1. the bread yeast to the oxidative damage hypersensitivity is characterized in that, this hypersensitivity bread yeast is disappearance Yap1cwp1cwp2pdr5snq2The bread yeast of five genes.
2. realize the preparation method of the described a kind of bread yeast to the oxidative damage hypersensitivity of claim 1, it is characterized in that this method comprises the following step:
(1), from wild bread yeast, extract genomic dna:
1a, get bread yeast, be inoculated in the YPD nutrient solution, at 30 ℃, under 200 rev/mins of conditions, shaking culture 16 hours obtains bread microzyme liquid;
Wherein: the weight percent of YPD nutrient solution prescription is: yeast extract paste 1%, and peptone 2%, glucose 2% the rest is water;
1b, get 1.5 milliliters of bread microzyme liquid, with 4000 rev/mins centrifugal 2 minutes, throw out is suspended from 200 microlitres and extracts in the damping fluid;
Wherein: the prescription that extracts damping fluid is: triton x-100 2%, and sodium laurylsulfonate 1%, 0.1 mole in sodium-chlor, ethylenediamine tetraacetic acid (EDTA) 1 mmole, Tri(Hydroxymethyl) Amino Methane Hydrochloride 10 mmoles the rest is water, and pH 8.0;
1c, add 0.3 gram pickling glass pearl, 100 microlitre phenol, 100 microlitre chloroforms again, shake after 3 minutes, with 12000 rev/mins centrifugal 10 minutes, the absorption supernatant liquid;
1d, add the dehydrated alcohol of 2 times of the liquid volumes of getting, left standstill 30 minutes at-20 ℃, with 12000 rev/mins centrifugal 15 minutes, remove supernatant, taking precipitate;
1e, throw out is dissolved in 20 microlitre sterilized waters, is pastoris genomic dna solution;
(2), make up the yap1 Δ:: hisG-URA3-hisG gene knockout assembly:
2a, at first utilize round pcr, by upstream primer YAP1-1:GGA TGC GTA AGG TCT TAA GAG and downstream primer YAP1-2:CAG TGG TTC TGC AGG TAC GG, the pastoris genomic dna fragment amplification of one section 2.5Kb is come out, wherein include open reading frame and the upstream homologous fragment of 0.2Kb and the downstream homologous fragment of 0.35Kb of YAP1;
Wherein: the condition of PCR is: 94 ℃ 5 minutes; 94 ℃ 45 seconds, 52 ℃ 1 minute, 72 ℃ 1 minute, 30 circulations; 72 ℃ 10 minutes;
2b, the pcr amplified fragment of the above-mentioned YAP1 of containing gene is cloned on the pGEM-T carrier, uses BamHIRestriction endonuclease and BstEIIRestriction endonuclease downcuts the segment of one section 1.8kb, contains the joint of BamHI restriction enzyme site in the replacement;
2c, usefulness BamHIRestriction endonuclease and The BglII restriction endonucleaseDowncut one section 3.8kb segment that contains hisG-URA3-hisG from the pNKY51 carrier, this segment is inserted into BamHI site in the step 2b joint;
2d, usefulness AflII and EcoRIEndonuclease digestion obtains YAP1 gene knockout assembly: the yap1 Δ:: hisG-URA3-hisG;
(3), with gene knockout assembly yap1 Δ:: hisG-URA3-hisG changes ultrahigh permeability bread yeast over to, and screening obtains yap1cwp1cwp2pdr5snq2 five gene knockout bread yeasts:
3a, ultrahigh permeability bread yeast is inoculated into 2 milliliters of YPD nutrient solutions, 30 ℃, under 200 rev/mins of conditions, shaking culture 16 hours;
3b, add 3 milliliters of YPD nutrient solutions again, 30 ℃, 200 rev/mins, cultivated 4 hours, obtain ultrahigh permeability bread yeast bacterium liquid;
3c, get 1.5 milliliters of ultrahigh permeability bread yeast bacterium liquid, with 2500 rev/mins centrifugal 2 minutes, taking precipitate;
3d, with throw out be suspended from 100 the milli rub/liter 400 microlitre Lithium Acetates in, with 2500 rev/mins centrifugal 2 minutes, taking precipitate;
3e, with throw out be suspended from 100 the milli rub/liter 100 microlitre Lithium Acetates in;
3f, smart single stranded DNA 2 microlitres of the salmon of 2.0 mg/ml were boiled 5 minutes, behind the ice bath with the yap1 Δ:: hisG-URA3-hisG gene knockout assembly 0.1～1 microgram joins in the Lithium Acetate of step 3e, and room temperature was placed 5 minutes;
3g, to add 280 microlitre concentration again be 50% Macrogol 4000;
3h, vibrate to mixing, 30 ℃ left standstill 30 minutes;
3i, add 39 microlitre dimethyl sulfoxide (DMSO), 42 ℃ of heat shocks 5 minutes;
Centrifugal 2 minutes of 3j, 4000 rev/mins, taking precipitate, and throw out is suspended from the 200 microlitre sterilized waters;
Centrifugal 2 minutes of 3k, 4000 rev/mins, taking precipitate is suspended from the 100 microlitre sterilized waters, be suspended from again in the non-selective substratum, grow overnight, and then be applied to 5-fluororotic acid selective medium flat board, cultivated 3 days in 30 ℃ of incubators, grow single bacterium colony, be Yap1cwp1cwp2pdr5snq2Five gene knockout bread yeasts, a kind of bread yeast just of the present invention to the oxidative damage hypersensitivity;
The wherein preparation of 5-fluororotic acid selective medium flat board: with yeast extract paste 1%, peptone 2%, glucose 2%, agar powder 2% the rest is water and places under 121 ℃ of high pressure sterilization 20 minutes, when treating that temperature drops to 50 ℃, adds 5-fluororotic acid 0.1%;
The wherein preparation of non-selective substratum: with yeast extract paste 1%, peptone 2%, glucose 2%, agar powder 2% the rest is water and places under 121 ℃ of high pressure sterilization 20 minutes.
CN 201010594171 2010-12-18 2010-12-18 Baker's yeast with high sensibility on oxidative damage and preparation method thereof Pending CN102120968A (en)

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CN102796735A (en) * 2012-07-27 2012-11-28 中国科学院水生生物研究所 Biosensor element box based on yeast DNA (deoxyribonucleic acid) damage response and application thereof
CN102965368A (en) * 2012-07-27 2013-03-13 中国科学院水生生物研究所 Super-sensitive yeast genotoxic carcinogen biosensor element box and its application
CN104099397A (en) * 2014-07-23 2014-10-15 中国科学院水生生物研究所 Method for detecting cytotoxicity of nano materials through yeast mutant strains
CN111778281A (en) * 2020-07-17 2020-10-16 四川省人民医院 Construction method and application of retina bipolar cytopathy model
WO2024074540A3 (en) * 2022-10-06 2024-08-08 Danmarks Tekniske Universitet Biocontainment strategies for in vivo applications of yeasts

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102796735A (en) * 2012-07-27 2012-11-28 中国科学院水生生物研究所 Biosensor element box based on yeast DNA (deoxyribonucleic acid) damage response and application thereof
CN102965368A (en) * 2012-07-27 2013-03-13 中国科学院水生生物研究所 Super-sensitive yeast genotoxic carcinogen biosensor element box and its application
CN102965368B (en) * 2012-07-27 2014-12-24 中国科学院水生生物研究所 Super-sensitive yeast genotoxic carcinogen biosensor element box and its application
CN104099397A (en) * 2014-07-23 2014-10-15 中国科学院水生生物研究所 Method for detecting cytotoxicity of nano materials through yeast mutant strains
CN111778281A (en) * 2020-07-17 2020-10-16 四川省人民医院 Construction method and application of retina bipolar cytopathy model
CN111778281B (en) * 2020-07-17 2021-04-23 四川省人民医院 Construction method and application of retina bipolar cytopathy model
WO2024074540A3 (en) * 2022-10-06 2024-08-08 Danmarks Tekniske Universitet Biocontainment strategies for in vivo applications of yeasts

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