CN102524177A - Method for constructing HRN/gpt delta transgenic mouse model - Google Patents
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Abstract
The invention discloses a method for constructing a liver cytochrome P450 enzyme function deficiency type gpt delta transgenic mouse model. The mouse model provided by the invention has all characteristics of a liver cytochrome P450 reductase-removed mouse and a gpt delta transgenic mouse, the research on the relationship between compound metabolism and mutagenesis can also be carried out, and the mouse model is especially applicable to researching compounds making extrahepatic tissue organs activated by liver CYP metabolism mutate.
Description
Technical field
The present invention relates to the construction method of animal model, in particular to a kind of liver cytochrome P 450 enzyme functional defect type
GptDelta (HRN/
GptDelta) construction method of transgene mouse model.
Background technology
The organism metabolism process both can have been detoxified to xenobiontics also can increase poison, and closely related to the carcinogenesis of body with compound.Liver cytochrome P 450 enzyme (being abbreviated as CYP) plays an important role in the metabolic process of body to exogenous compounds, but it is very limited to can be used for disclosing the research tool of its carcinogenesis.
At present, research liver P450 enzyme mainly concentrates on the experiment in vitro the metabolism of exogenous compounds, comprises the application of liver microsomes, primary hepatocyte, each hypotype high expressing cell strain of CYPs and recombinase.Although these in-vitro methods have very big using value; But,, make above-mentioned experiment in vitro still be difficult to accurately predicting exogenous compounds actual metabolic pathway (Friedberg T in vivo because the absorption approach of exogenous compounds is different, the existence of the complicated factors such as the outer CYPs effect of existence, kidney scavenging action and liver of organic the moon (sun) ion transport body in the tissue; Pritchard MP; Bandera M, Hanlon SP, Yao D; McLaughlin LA; Ding S, Burchell B, Wolf CR. Merits and limitations of recombinant models for the study of human P450-mediated drug metabolism and toxicity:an intralaboratory comparison. Drug Metab Rev. 1999.31 (2): 523-44.).In the metabolism research, have a plurality of P450 enzyme hypotype knock-out mice models at present in vivo, these models have been widely used for studying specificity P450 enzyme hypotype in the metabolism of exogenous compounds and the effect in the toxicity.Yet CYP has multiple hypotype usually, and each hypotype function class seemingly, and substrate exists and intersects, and makes the gene knockout animal model of single hypotype be difficult to accurately estimate the influence of P450 enzyme to exogenous compounds toxicity.Therefore, the model that makes all CYPs all lose function will be a very useful instrument.NADPH-cytochrome P450 reductase (CPR/POR) is the oxidoreduction partner of all microsome P450 enzymes; It provides first required electronics of P450 oxydasis reduction reaction; Based on this principle, people such as Jun Gu developed a liver specificity P450 reductase-
CprGene knock-out mice model (
HEpatic
REductase
NUll, HRN is designated hereinafter simply as liver specificity CPR KO mouse model), the disappearance of this gene expression will suppress activity (the Wu L of all liver P450 enzymes; Gu J, Weng Y, Kluetzman K; Swiatek P, Behr M, Zhang QY; Zhuo X, Xie Q, Ding X. Genesis. 2003 Aug; 36 (4): 177-81.).This model utilizes the Cre/LoxP system to knock out (conditional knock) through condition and obtains, and mouse is liver in maturation
CprGene is knocked out (being born after 2 months) gradually, finally make liver P450 enzymic activity lose, and its hetero-organization is unaffected.
Most carcinogenic substances have genetoxic, promptly damage genetic material (DNA or chromosome), cause gene mutation and then carcinogenic.Genotoxic evaluation is the effective ways of prediction carcinogenic, and the object of its detection comprises the change of gene mutation, chromosome damage, reorganization and number that dna damage and dna damage cause etc.Estimate employing standard combination test usually and carry out, like Salmonella reversion test, mammalian cell gene mutation test and the transgenic animal sudden change detection test etc. of the Using Comet Assay of estimating dna damage, the micronucleus test of estimating chromosome damage and chromosomal aberration test, evaluation gene mutation.Compare with experiment in vitro, vivo experiment method false positive incidence is lower, and has the advantage of considering the absorption relevant with human use, metabolism, distribution and drainage, so the application of experiment in the body comes into one's own day by day.Existing In vivo assay Cells is less, the measurable carcinogenic of micronucleus test in the classical body, but this method is only estimated marrow and can not be predicted the toxicity target organ.Transgenic animal are very important research tools, can detect any required intraorganic gene mutation, the mechanism analysis that can also suddenly change.
Transgenic animal sudden change detection model always mainly contains (J A Gossen such as Muta Mouse and Big Blue Mouse/Rat; W J de Leeuw; C H Tan, E C Zwarthoff, F Berends; P H Lohman; D L Knook, and J Vijg. Efficient rescue of integrated shuttle vectors from transgenic mice:a model for studying mutations in vivo, Proc. Natl. Acad. Sci. U.S.A
. 1989; 86:7971-7975.Kohler SW, Provost GS, Kretz PL; Fieck A; Sorge JA, Short JM. The use of transgenic mice for short-term, in vivo mutagenicity testing. Genet Anal Tech Appl. 1990 Dec; 7 (8): 212-8.).But the common defects that above-mentioned model has be can only test point the deletion mutation of sudden change and DNA small fragment, and the deletion mutation of fragment just is difficult to be detected greatly.1996, people such as Nohmi set up new transgenic animal model
GptDelta transgenic mice (Nohmi T, Katoh M, Suzuki H; Matsui M, Yamada M, Watanabe M; Suzuki M, Horiya N, Ueda O; Shibuya T, Ikeda H, Sofuni T.A new transgenic mouse mutagenesis test system using Spi-and 6-thioguanine selections. Environ Mol Mutagen. 1996; 28 (4): 465-70.), the characteristics of this model are the sudden changes that not only can detect bases such as comprising point mutation, frameshift mutation, base replacement in the Different Organs of mouse, can also detect dna fragmentation and lack on a large scale.In addition,
GptThe length of gene is 456 bp, helps further through order-checking mechanism analysis being carried out in sudden change.
Confirm that the relation between mutagenesis detects in liver cytochrome P 450 and the body has great significance to the genetic predisposition prediction and the prevention of compound.In two kinds of mouse models of this that has at present, liver specificity CPR KO mouse model can not carry out vivo mutations and detect,
GptThough the delta transgenic mice can be done vivo mutations and detect, and can't investigate liver cytochrome P 450 role therein.Therefore, the animal model that has only foundation to possess two specific characters simultaneously just can address this problem.
Summary of the invention
To deficiency of the prior art, the inventor is devoted to study the animal model that possesses two specific characters simultaneously.Thus, the present invention is two kinds of animal patterns of using
GptThe delta transgenic mice is hybridized with the liver specificity CPR KO mouse with identical C57BL/6J genetic background, through detecting genotype, select required filial generation animal to carry out mating repeatedly, has finally obtained liver specificity CPR deficiency
GptDelta transgenic mice new model is liver P450 enzyme functional defect type
GptThe delta transgenic mice (
HEpatic
REductase
NUll
GptThe delta transgenic mice abbreviates HRN/ as
GptThe delta transgenic mice).And it is unsuccessful that the present invention has also overcome hybridization in repetition test, and filial generation can not be survived or genotype is not inconsistent with expection, and make the technical barrier that each gene of mouse after the hybridization can genetic stability.
Therefore, an object of the present invention is to provide a kind of liver cytochrome P 450 enzyme functional defect type
GptThe delta transgene mouse model.
Another object of the present invention provides a kind of liver cytochrome P 450 enzyme functional defect type
GptThe construction method of delta transgene mouse model.
A further object of the present invention provides the application in the relation between research liver cell P450 enzyme and compound mutagenesis of this model.
For realizing above-mentioned purpose, the invention provides a kind of liver cytochrome P 450 enzyme functional defect type
GptThe delta transgene mouse model.
According to another object of the present invention, the invention provides a kind of method that makes up above-mentioned model, this method may further comprise the steps:
(1) select for use respectively liver specificity CPR KO mouse with
GptThe delta transgenic mice is hybridized, and obtains F1 generation:
Cpr Loxp+/-
Cre +/- Gpt +/-With
Cpr Loxp+/-
Cre -/- Gpt +/-Mouse genotypes.Because two kinds of mouse genetic backgrounds are consistent, all are C57BL/6J strain mouse, so there is not the genetic background difference problem in generation mice.
(2) with in F1 generation
Cpr Loxp+/-
Cre +/- Gpt +/-Genotypic female mouse and male mouse hybridize again, obtain F2 generation:
Cpr Loxp+ /+
Cre +/- Gpt + /+With
Cpr Loxp+ /+
Cre -/- Gpt + /+Mouse genotypes.
(3) with in F2 generation
Cpr Loxp+ /+
Cre +/- Gpt + /+The male mouse of genotype with
Cpr Loxp+ /+
Cre -/- Gpt + /+The female mouse of genotype carries out mating, produces more F3 generation
Cpr Loxp+ /+
Cre +/- Gpt + /+With
Cpr Loxp+ /+
Cre -/- Gpt + /+Mouse genotypes.
(4) with in F3 generation
Cpr Loxp+ /+
Cre +/- Gpt + /+The male mouse of genotype with
Cpr Loxp+ /+
Cre -/- Gpt + /+The female mouse of genotype is the liver cytochrome P 450 enzyme functional defect type that can normally be bred of post-coitum again
GptThe delta transgene mouse model.
Above-mentioned steps all needs to utilize the specific gene amplimer to carry out genotype identification to the mouse that is obtained.
According to another object of the present invention, the invention provides the application in the relation between research liver cell P450 enzyme and compound mutagenesis of above-mentioned mouse model.
Liver cytochrome P 450 enzyme functional defect type of the present invention
GptThe delta transgene mouse model is following with the beneficial effect of compared with techniques in the past:
At first, compared with prior art, liver cytochrome P 450 enzyme functional defect type of the present invention
GptThe delta transgene mouse model be through with liver specificity CPR KO mouse with
GptThe delta transgenic mice obtains behind a series of hybridization and genotype screening, and the characteristic that it has two kinds of parental generation models concurrently is the indispensable technological means of relation between research liver P450 enzyme and the compound mutagenesis.
Secondly, new model of the present invention also possesses whole application of two kinds of parental generation models:
A. the application that has liver specificity CPR KO mouse aspect: whether compound passes through metabolism and the effect of liver CYP; The effect of CYP in the compound metabolism in outer its hetero-organization internal organs of liver; CYP totally different in endogenous and exogenous compounds metabolism in liver microsomes and the mitochondria; Liver HO enzyme function changes the relevant metabolic alterations in back; Also can be used for research can CYP metabolism in liver, but in the extrahepatic tissue internal organs toxigenous compound; Be used to study the compound that hepatotoxicity wind agitation strengthens after the liver CYP metabolism.
B. have
GptThe application of delta transgenic mice aspect: because λ EG10 DNA is present in the genome of mouse, therefore can do detection in Gene Mutation and analysis to the internal organs of organizing of mouse whole body, but test point sudden change and deletion mutation; And compound is to the comparative studies of the mutagenesis of different tissues organ.
Once more, this new model not only have liver cytochrome P 450 reductase knock-out mice with
GptAll characteristics of delta transgenic mice, but also can carry out the research that concerns between compound metabolism and the mutagenesis, be particularly suitable for the research that those can be formed the compound of sudden change by liver CYP metabolism activation extrahepatic tissue organ.
In addition, in the process that new model makes up, it is unsuccessful that the present invention has also overcome hybridization, the difficult problem that filial generation can not be survived or genotype and expection are not inconsistent, this be because liver specificity CPR KO mouse with
GptThe delta transgenic mice all is C57 BL/6J strain mouse, and genetic background is more consistent, so the hybridization chance of success is bigger; Liver specificity CPR KO mouse with
GptDelta transgenic mice fertility is normal, in the new mouse genotypes that the hybridization back forms
CprNon-interaction action between gene knockout mechanism and the λ EG10 sequence, so generation mice not only can be survived but also fertility is normal.And, since liver specificity CPR KO mouse with
GptMany different gene type mouse may occur after the hybridization of delta transgenic mice, the genotype screening process that the present invention adopted can effectively solve this difficult problem.Further, the inventor exists
Cpr Loxp Gene,
CreGene with
GptGene can the basis of genetic stability on, hybridize mice obtains
Cpr Loxp Gene with
GptCarry out normal breeding again after the sub-mouse of gene pure, so just guarantee in the new model mouse
Cpr Loxp Gene,
CreGene with
GptGene is genetic stability in each generation.
Description of drawings
Fig. 1 is a liver cytochrome P 450 enzyme functional defect type
GptThe flow chart that the delta transgene mouse model makes up.
Fig. 2 be F1 for the genotypic PCR qualification result of mouse, A is for having
CreThe mouse of gene, wherein,
Cpr Loxp Genetic heterozygosis expands and 500 bp and 600 bp, two bands,
CreGene expands and 300 bp, one band,
GptGenetic heterozygosis expands and 360 bp (F-R1) and 967 bp (F-R2), two bands; Be nothing among the B
CreDna murine, wherein,
Cpr Loxp Genetic heterozygosis is sub, expands 500 bp and 600 bp, two bands,
GptGenetic heterozygosis expands and 360 bp (F-R1) and 967 bp (F-R2), two bands.
Fig. 3 be F2 for the genotypic PCR qualification result of mouse, A is for having
CreThe mouse of gene, wherein,
Cpr Loxp Gene pure expands and 600 bp, one band,
CreGene expands and 300 bp, one band,
GptGene pure expands and 967 bp (F-R2) band; B is not for having
CreDna murine,
Cpr Loxp Gene pure expands and 600 bp, one band,
GptGene pure expands and 967 bp (F-R2) band.
Fig. 4 is according to the liver specificity CPR deficiency in F2 generation in one embodiment of the present invention
GptMain organs qualification result in the delta transgenic mice.A does
Cpr Loxp Gene with
CreGene, B does
GptGene, wherein 967 bp are the PCR result of primers F-R2 of using in the genotype identification, and 600 bp are primers
Gpt-F with
Gpt-R qualification result; L: liver; K, kidney; St, stomach; B, bladder; Sp, spleen.
Embodiment
Below in conjunction with embodiment the present invention is done further elaboration, following embodiment is only described the present invention by way of example.Clearly, those of ordinary skills can carry out various accommodations and modification to the present invention in scope of the present invention and essence.Need be appreciated that, this invention is intended to be encompassed in the accommodation and the modification
that comprise in the appended claims.
The inventor is through deeply and extensive studies, made up a kind of inheritance stability, liver cytochrome P 450 enzyme functional defect type that phenotype is stable
GptThe delta transgene mouse model is specially: select for use the male liver specificity CPR of parental generation KO mouse and parental generation female respectively
GptThe delta transgenic mice is hybridized (F0 generation).With F1 in generation
Cpr Loxp+/-
Cre +/- Gpt +/-Genotypic female mouse and male mouse hybridize and obtain F2 generation
Cpr Loxp+ /+
Cre +/- Gpt + /+With
Cpr Loxp+ /+
Cre -/- Gpt + /+Mouse genotypes is genotypic mouse required for the present invention, a large amount of screenings of this process need.Then, with in F2 generation
Cpr Loxp+ /+
Cre +/- Gpt + /+Male mouse with
Cpr Loxp+ /+
Cre -/- Gpt + /+Female mouse carries out expanding propagation through mating, thereby produces more F3 generation
Cpr Loxp+ /+
Cre +/- Gpt + /+With
Cpr Loxp+ /+
Cre -/- Gpt + /+Mouse genotypes.With F3 generation
Cpr Loxp+ /+
Cre +/- Gpt + /+Male mouse with
Cpr Loxp+ /+
Cre -/- Gpt + /+Female mouse is post-coitum again, and F4 is carried out the identified gene type for sub-mouse, but the result shows three kinds of equal genetic stabilities of gene in the sub-mouse of gained, promptly explains from F3 for can normally breeding.Entire making process all need utilize the specific gene amplimer to carry out genotype identification.
The practical implementation process:
1. F0 is for hybridization:
The male liver specificity CPR of parental generation KO mouse and parental generation are female
GptThe delta transgenic mice is hybridized, and produces F1 generation.Cut toe identified gene type at F1 4 weeks for generation mice birth back.
Extract genomic DNA:
The Ep that the toe of cutting is put into 1.5mL manages, and adds the DNA digestion buffer solution that 0.4mL contains the 0.4mg/mL Proteinase K, puts into water-bath then, 55 ℃ of digested overnight (digestion time is more than 16h).Next day, treat mouse toe digestion fully after, in each Ep pipe, add 0.4 mL phenol chloroform (pH 8.0), turn upside down and mix 50-60 time, 15,4 ℃ of 000g are centrifugal 20 minutes.Get 300 μ L supernatants and insert in other 1 new 1.5 mL Ep pipe,, put upside down gently up and down and mix 5-10 time, centrifugal 5 minutes of 12,000 rpm to wherein adding 750 μ L ice absolute ethyl alcohol.Abandon supernatant, add ice 75% ethanol 200 μ L and wash 2 times.The Ep pipe that will contain genomic DNA tips upside down on blotting paper last 30 minute, dry DNA.In the Ep pipe, add 80～100 μ L TE buffer solutions, room temperature was placed 2 hours, and DNA is fully dissolved.
Detect genotype:
Genome with extracting is PCR, detects F1 for the mouse genotype.
Primer sequence and product size:
Cpr Loxp Gene primer:
Upstream primer 1 (F): 5 '-TACAATGGACCAGGCTCTGC-3 '
Downstream primer 2 (R): 5 '-AAGAGGGACAAAGAGCACC-3 '
The PCR product: isozygoty, 600bp (contains
Loxp), a band; Heterozygosis 500bp (does not contain
Loxp) and 600bp (contain
Loxp) each band.
CreGene primer:
Upstream primer 3 (F): 5 '-GGATTT CCGTCTCTGGTGTAGC-3 '
Downstream primer 4 (R): 5 '-CATTGCCCCTGTTTCACTATCC-3 '
PCR product: 300bp, a band.
GptGene primer:
Upstream primer 5 (F): 5 '-GTTGTACTTCCAACCATGCCAAAG-3 '
Downstream primer 6 (R1): 5 '-CAGAAATCATTCCAGGTCCTTGC-3 '
Downstream primer 7 (R2): 5 '-GTTCATCTGCTTTATGGGCAAGAG-3 '
Upstream primer 8 (
Gpt-F): 5 '-GCGCAACCTATTTTCCCCTCGA-3 '
Downstream primer 9 (
Gpt-R): 5 '-TGGAAACTATTGTAACCCGCCTG-3 '
The PCR product:Homozygote, 967bp (the PCR product of primers F and primer R1), a band; Heterozygote is 360bp (the PCR product of primers F and primer R1) and 967bp (the PCR product of primers F and primer R2), two bands; Wild type has only 360bp (the PCR product of a primers F and primer R1) band.
Gpt-F with
Gpt-R product is 600bp, the main evaluation
GptGene.
Program setting:
Cpr Loxp With
CreThe gene PCR program setting: 94 ℃ of preparatory sex change 3 minutes, 94 ℃ 30 seconds, 64 ℃ 30 seconds, 72 ℃ 30 seconds, totally 36 circulations, 72 ℃ were extended 4 ℃ of preservations 7 minutes then.
GptThe gene PCR program setting: 94 ℃ of preparatory sex change 4.5 minutes, 94 ℃ 30 seconds, 58 ℃ 30 seconds, 72 ℃ 1 minute, totally 30 circulations, 72 ℃ were extended 4 ℃ of preservations 5 minutes then.
Product is identified:
Getting 10 μ L PCR products after PCR finishes identifies in 1.5% agarose gel electrophoresis.Amplification is seen shown in Figure 1, and A has shown
CreThe genotype of dna murine, wherein, the 1st swimming lane is 100 bp DNA ladder Marker, the 2nd swimming lane does
Cpr Loxp Genetic heterozygosis, 500 bp and 600 bp, the 3rd swimming lane does
CreGene, 300 bp, the 4th, 5 swimming lanes do
GptGenetic heterozygosis 360 bp (F-R1) and 967 bp (F-R2).B shows nothing among Fig. 1
CreThe genotype of gene (300 bp) mouse, wherein, the 1st swimming lane is 100 bp DNA ladder Marker, the 2nd swimming lane does
Cpr Loxp Genetic heterozygosis, 500 bp and 600 bp, the 4th, 5 swimming lanes do
GptGenetic heterozygosis 360 bp (F-R1) and 967 bp (F-R2).
The F1 that the hybridization back forms has two kinds for genotype:
Cpr Loxp+/-
Cre +/- Gpt +/-With
Cpr Loxp+/-
Cre -/- Gpt +/-
Generation hybridization:
With F1 generation male and female
Cpr Loxp+/-
Cre +/- Gpt +/-The mouse mating, and to F2 generation sub-mouse according to aforesaid way identified gene type.It is more that the mensuration result shows that F2 genotype occurs for sub-mouse, and screening also keeps
Cpr Loxp+ /+
Cre +/- Gpt + /+With
Cpr Loxp+ /+
Cre -/- Gpt + /+Mouse genotypes.
As shown in Figure 2, A has shown
CreThe genotype of dna murine, wherein, the 1st swimming lane is 100 bp DNA ladder Marker, the 2nd swimming lane does
Cpr Loxp Gene pure, 600 bp, the 3rd swimming lane does
CreGene, 300 bp, the 4th, 5 swimming lanes do
GptGene pure does not have 360 bp (F-R1) gene outcome, has only 967 bp (F-R2).B shows nothing among Fig. 2
CreThe genotype of gene (300 bp) mouse, wherein, the 1st swimming lane is 100 bp DNA ladder Marker, the 2nd swimming lane does
Cpr Loxp Gene pure, 600 bp, the 4th, 5 swimming lanes do
GptGene pure does not have 360 bp (F-R1) gene outcome, has only 967 bp (F-R2).
For expanding propagation:
With F2 generation male
Cpr Loxp+ /+
Cre +/- Gpt + /+Mouse and female
Cpr Loxp+ /+
Cre -/- Gpt + /+The mouse mating, and to F3 generation sub-mouse according to aforesaid way identified gene type.Because F2 is in the sub-mouse
Cpr Loxp+ /+
Cre +/- Gpt + /+Mouse with
Cpr Loxp+ /+
Cre -/- Gpt + /+The mouse proportion is less, therefore need be with two kinds of mouse post-coitum expanding propagation.
For normal breeding:
With F3 generation male
Cpr Loxp+ /+
Cre +/- Gpt + /+Mouse and female
Cpr Loxp+ /+
Cre -/- Gpt + /+The mouse mating, and to F4 generation sub-mouse according to aforesaid way identified gene type.The result shows, gets final product normal breeding from F3 generation, but three kinds of equal genetic stabilities of gene.
Sequence table
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< 120>construction method of HRN/gpt delta transgene mouse model
<130> DI10-2039-XC68
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Claims (4)
1. liver cytochrome P 450 enzyme functional defect type
GptThe construction method of delta transgene mouse model is characterized in that, may further comprise the steps:
Select for use respectively liver specificity CPR KO mouse with
GptThe delta transgenic mice is hybridized, and obtains F1 generation:
Cpr Loxp+/-
Cre +/- Gpt +/-With
Cpr Loxp+/-
Cre -/- Gpt +/-Mouse genotypes;
With F1 in generation
Cpr Loxp+/-
Cre +/- Gpt +/-Genotypic female mouse and male mouse hybridize again, obtain F2 generation:
Cpr Loxp+ /+
Cre +/- Gpt + /+With
Cpr Loxp+ /+
Cre -/- Gpt + /+Mouse genotypes;
With F2 in generation
Cpr Loxp+ /+
Cre +/- Gpt + /+The male mouse of genotype with
Cpr Loxp+ /+
Cre -/- Gpt + /+The female mouse of genotype carries out mating, produces more F3 generation
Cpr Loxp+ /+
Cre +/- Gpt + /+With
Cpr Loxp+ /+
Cre -/- Gpt + /+Mouse genotypes; And
With F3 in generation
Cpr Loxp+ /+
Cre +/- Gpt + /+The male mouse of genotype with
Cpr Loxp+ /+
Cre -/- Gpt + /+The female mouse of genotype post-coitum again can obtain the liver cytochrome P 450 enzyme functional defect type that can normally breed
GptThe delta transgene mouse model.
2. the method for claim 1 is characterized in that, said CPR KO mouse with
GptThe delta transgenic mice has identical C57BL/6J genetic background.
3. the method for claim 1 is characterized in that, said each step further comprises utilizes the specific gene amplimer to carry out genotype identification to the mouse that is obtained.
4. liver cytochrome P 450 enzyme functional defect type as claimed in claim 1
GptThe delta transgene mouse model is the application in the relation between research liver cell P450 enzyme and compound mutagenesis.
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| CN106222187A (en) * | 2016-08-03 | 2016-12-14 | 赣南医学院第附属医院 | Transgene carrier, transgene mouse model and the construction method thereof of inducible c myc process LAN |
| CN110506702A (en) * | 2019-08-09 | 2019-11-29 | 浙江大学 | A kind of method for building up of novel model of cardiac arrest |
| CN111838066A (en) * | 2019-07-12 | 2020-10-30 | 上海吉辉实验动物饲养有限公司 | Spontaneous mutation hairless SHJHhr/hrEarly-aging mouse animal model and cultivation method thereof |
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