KR101811518B1 - Method for regulating oxidative stress resistant of Drosophila - Google Patents

Abstract

The present invention relates to a method for controlling the oxidative stress resistance of a fruit fly comprising the step of regulating the expression level of the Art1 gene, an oxidative stress resistant gene derived from a fruit fly, a method for controlling the oxidative stress resistance of the fruit fly by introducing and overexpressing the Art1 gene, And a method for screening an oxidative stress-resistant preparation using the transformed Drosophila and the transgenic Drosophila. Using the method for controlling the oxidative stress resistance of the fruit flies provided by the present invention, it is possible to more precisely study the in vivo action mechanism of the Art1 gene directly related to the lifespan, There will be.

Description

The present invention relates to a method for regulating oxidative stress resistance of fruit flies,

The present invention relates to a method for controlling the oxidative stress resistance of fruit flies, and more particularly, the present invention relates to a method for controlling the oxidative stress resistance of a fruit fly comprising the step of regulating the expression level of the Art1 gene as an oxidative stress- A method for screening an oxidative stress resistant agent using the transformed Drosophila and an Art 1 gene introduced and overexpressed and having increased resistance to oxidative stress.

Aging is a natural life phenomenon that can not be avoided by all living organisms, and means the entire process in which physiological function decreases over time and eventually leads to death. A representative hypothesis explaining the mechanism of aging is the aging promotion theory based on oxidative stress due to accumulation of active oxygen. Life is constantly resistant to external stimuli and internal stress. It is the theory that aging will accelerate aging by this loss of resistance. There have been many reports that the increase in resistance to stress prolongs the life span and has an anti-aging effect. In this respect, efforts to apply stress-resistant genes to anti-aging studies are still continuing. Particularly in the fields of medicine and research, the Drosophila model is widely used, which is easy to control gene insertion and gene expression, has a short average life span, and is capable of human gene insertion and function studies.

Among the above stresses, oxidative stress refers to a state in which redox re-regulation is required by increasing reactive oxygen species (ROS) and reactive nitrogen species (RNS) in a cell. In other words, active oxygen and active nitrogen are produced in the normal state of mitochondria, peroxisome, and cytoplasm, and can be generated through various mechanisms such as the action of antibiotics, anticancer agents, toxic compounds, and other transition metals These reactive oxygen species act as mediators of physiological and pathological processes in vivo, leading to chronic and acute inflammatory diseases, and in high concentrations in environmentally stressed conditions, they kill cells. However, since various defense mechanisms such as antioxidant enzymes are present in the cells, such active oxygen and active nitrogen can be removed and the normal state can be maintained. However, if the defense mechanisms are damaged due to a specific cause, the cells are damaged by oxidative stress. In other words, lipid peroxidation, oxidation of amino acids (especially cysteine), cross-linking of proteins, DNA damage, DNA strand breakdown, protein segmentation and cell death due to apoptosis as well as mutation and cancer, aging, cardiovascular disease, It is known to cause more than 100 diseases such as immune diseases and neurodegenerative diseases.

Studies on the genes associated with such oxidative stress have been actively carried out to investigate the mechanism of defense in the body. For example, Korean Patent Publication No. 2003-0076058 discloses a method of transforming aNDPK2 gene into a plant to make it resistant to stress. Korean Laid-Open Patent Application No. 2015-0055582 discloses a gene specific to environmental stress of poultry m-calpain, Caspase 3 and Cathepsin B are disclosed. However, studies on regulating in vivo activities such as aging or lifespan using these stress-related genes have not been disclosed until now.

Under these circumstances, the present inventors have made intensive researches to develop a method for regulating in vivo activity such as aging or lifespan using a stress-related gene directly related to aging or lifespan. As a result, the expression level of Art1 gene derived from Drosophila The present inventors have confirmed that the resistance to oxidative stress can be regulated, and in particular, when the expression of Art1 gene is suppressed, the lifespan of fruit flies is reduced.

It is an object of the present invention to provide a method for controlling the oxidative stress resistance of a fruit fly comprising the step of regulating the level of expression of the Art1 gene in a fruit fly.

Another object of the present invention is to provide a transformed Drosophila having an increased resistance to oxidative stress by introducing and overexpressing the Art1 gene.

It is another object of the present invention to provide a method for screening an oxidative stress resistant agent using the transgenic flies.

The present inventors have paid attention to fruit flies while carrying out various studies in order to develop a method of regulating in vivo activity such as aging or life span by using stress related genes directly related to aging or lifespan. Drosophila is an experimental animal to directly study the effect of stress genes because it is easy to breed, has a short generation time, can obtain many offspring in a short time, has easy genetic manipulation, and has the highest level of gene analysis And to study the effect of deletion or overexpression of Art1 gene derived from Drosophila. As a result, it was confirmed that when the Art1 gene was deleted in Drosophila, the life span was specifically reduced in female Drosophila. Therefore, it was confirmed that resistance to oxidative stress can be directly regulated by analyzing the resistance to stress related to the life span and the expression of Art1 gene. The results that the Art1 gene can directly control the oxidative stress of the fruit fly and thus directly affect the fruit fly life have not been reported so far and have been first identified by the present inventor.

In order to achieve the above object, the present invention provides, as one embodiment, a method for controlling the oxidative stress resistance of a fruit fly comprising the step of regulating the expression level of the Art1 gene in a fruit fly.

The method of controlling the oxidative stress resistance of the fruit flies provided by the present invention shows a sex specificity. In the case of the male fruit flies, the resistance to the oxidative stress is not changed even when the expression level of the Art1 gene is regulated, Increasing the expression level of the Art1 gene may increase the resistance to oxidative stress and decrease the expression level of the Art1 gene, which may shorten the life span of the fruit flies.

The term "Drosophila " of the present invention refers to an insect belonging to one insect belonging to the insect insect double-headed tree of the arthropods, and generally includes D. melanogaster, D. virilis and the like. The fruit flies are easy to breed and have a short generation time, so that a large number of offspring can be obtained in a short time, and they are used as experimental animals for study of genetics. In particular, yellow flies are one of the most developed animals with genetic interpretation and are used as a major experimental animal in various fields including genetics, embryology, behavioral science, time biology, ecology and so on.

The term "Art1 (Arginine methyltransferase 1) gene" of the present invention is homologs of PRMT (Protein arginine methyltransferase), which is involved in histone modification and is a protein arginine methylating enzyme known to modulate hormone receptor Means a gene encoding arginine methyltransferase 1. In the present invention, the Art1 gene having the nucleotide sequence of SEQ ID NO: 1 was used.

The expression level of the Art1 gene can be regulated by decreasing or increasing the expression level of the Art1 gene.

As a method for decreasing the expression level of the Art1 gene, there is a method of damaging the Art1 gene itself by deletion, substitution, insertion, etc., or by inactivating the transcriptional regulatory element of the Art1 gene, or by using siRNA or miRNA, Or the like may be used. For example, when an expression vector containing the damaged Art1 gene is introduced into a fertilized egg of a fruit fly, the damaged Art1 gene contained in the expression vector introduced into the fertilized egg is replaced with the Art1 gene contained in the genome DNA of the fruit fly, It can not be normally expressed or its expression can be inhibited.

As a method of increasing the expression level of the Art1 gene, a method of introducing an expression vector containing a normal Art1 gene into a fertilized egg of a fruit fly can be used. For example, when an expression vector containing a normal Art1 gene is introduced into a fertilized egg of Drosophila, the normal Art1 gene contained in the expression vector introduced into the fertilized egg is inserted into genomic DNA of Drosophila, or in the form of a plasmid in the cytoplasm The Art1 gene can be expressed and the expression level of the Art1 gene can be increased.

The term "expression vector" of the present invention means a recombinant vector capable of expressing a peptide of interest in a suitable host cell, including a necessary regulatory element operatively linked to the expression of the gene insert. Expression vectors of the present invention include those elements normally present in a suitable expression vector, including expression regulatory elements such as promoters, operators, and initiation codons. The initiation codon and termination codon are generally considered to be part of the nucleotide sequence encoding the polypeptide and must be operative in the individual when the gene product is administered and in the coding sequence and in frame. The promoter of the vector may be constitutive or inducible.

It may also contain a signal sequence for the release of the fusion polypeptide to facilitate the separation of the protein from the cell culture. A specific initiation signal may also be required for efficient translation of the inserted nucleic acid sequence. These signals include the ATG start codon and adjacent sequences. In some cases, an exogenous translational control signal, which may include the ATG start codon, should be provided. These exogenous translational control signals and initiation codons can be of various natural and synthetic sources. Expression efficiency can be increased by the introduction of suitable transcription or translation enhancers.

The expression vector may be any conventional expression vector. For example, plasmid DNA, phage DNA, etc. may be used. Specific examples of plasmid DNA include commercial plasmids such as pUC18, pIDTSAMRT-AMP. Other examples of plasmids that can be used in the present invention include plasmids derived from Escherichia coli (pYG601BR322, pBR325, pUC118 and pUC119), Bacillus subtilis-derived plasmids (pUB110 and pTP5), and yeast-derived plasmids (YEp13, YEp24, YCp50). Specific examples of phage DNA include lambda-phages (Charon4A, Charon21A, EMBL3, EMBL4, lambda gt10, lambda gt11 and lambda ZAP). In addition, animal viruses such as retrovirus, adenovirus or vaccinia virus, insect viruses such as baculovirus may also be used. Since the expression amount of the protein and the expression of the expression of the expression vector differ depending on the host cell, the host cell most suitable for the purpose may be selected and used.

In the present invention, the expression vector can be interpreted to be an expression vector for inactivation or introduction of a normal Art1 gene into the embryo of a fruit fly.

According to one embodiment of the present invention, a transgenic Drosophila having an Art1 gene deficient was prepared (Fig. 1), and the lifespan of the transgenic Drosophila isolated with the Art1 gene was measured. As a result, Mean life span was reduced, while the average life span was not reduced even when the Art1 gene was deleted in the male Drosophila (Figs. 2a and 2b). In order to examine the effect of overexpression of the Art1 gene, a transgenic Drosophila having overexpression of the Art1 gene was prepared, and the stress resistance of the transgenic Drosophila expressing the Art1 gene deficient or overexpressed was analyzed. As a result, (Fig. 3a and 3b) and the starvation stress resistance (Fig. 3c and 3d), the overexpression of the Art1 gene did not significantly affect the resistance to oxidative stress. However, when the Art1 gene was deleted, And increased resistance when overexpressed (FIGS. 3e and 3f).

In another aspect, the present invention provides a transgenic fruit fly having an increased resistance to oxidative stress by introducing and overexpressing the Art1 gene.

As described above, since the Art1 gene provided by the present invention enhances resistance to oxidative stress when overexpressed in Drosophila, the transgenic Drosophila into which the Art1 gene is introduced is suitable for carrying out studies on oxidative stress Can be used as a model. For example, the transgenic flies can be screened for substances capable of inducing oxidative stress, screened for substances capable of reducing oxidative stress, or tested for gene pathways involved in oxidative stress transmission Which can be used as an experimental model.

In another aspect, the present invention provides a method for screening a substance exhibiting resistance to oxidative stress using an inactivated Art1 gene-inactivated transgenic flora and an overexpressed Art1 gene in a transgenic flora.

Specifically, the screening method of the oxidative stress resistant preparation of the present invention comprises the steps of: (a) administering an oxidative stress resistant candidate agent to a transformed Drosophila having an Art1 gene inactivated; (b) measuring the survival rate of each fruit fly over time by adding oxidative stress to the transformed Drosophila in which the Art1 gene is inactivated and the transformed Drosophila in which the Art1 gene is overexpressed; And (c) determining a candidate substance administered to the transformed Drosophila in which the Art1 gene having a higher survival rate than the transgenic Drosophila that overexpressed the Art1 gene is inactivated, as an oxidative stress resistant agent.

The screening method comprises adding an oxidative stress-resistant candidate substance to a transformed Drosophila in which the Art1 gene is inactivated, which has not been increased in resistance to oxidative stress, thereby increasing the oxidative stress resistance of the transformed Drosophila that overexpresses the Art1 gene If it is higher than the stress resistance, the candidate substance is selected as an oxidative stress resistant preparation and selected. Using this method, it is possible to primarily select a formulation with a significantly stronger oxidative stress resistance, which can reduce unnecessary trial and error in the development of subsequent oxidative stress resistant preparations.

Using the method for controlling the oxidative stress resistance of the fruit flies provided by the present invention, it is possible to more precisely study the in vivo action mechanism of the Art1 gene directly related to the lifespan, There will be.

FIG. 1 is a graph showing the results of measuring the expression level of Art1 gene according to the treatment of RU486 with respect to male and female fruit flies.
2A is a graph showing the results of measurement of the lifespan of female transgenic flies deficient in Art1 gene.
FIG. 2B is a graph showing the results of measurement of the lifespan of a male transgenic flora deficient in Art1 gene. FIG.
FIG. 3A is a graph showing the results of comparing the resistance to high temperature stress of female transgenic flies deficient in Art1 gene. FIG.
FIG. 3B is a graph showing the results of comparing the resistance to high temperature stress of the female transgenic flies overexpressing Art1 gene.
FIG. 3C is a graph showing the results of comparing the resistance to starvation stress of female transgenic flies deficient in Art1 gene. FIG.
FIG. 3D is a graph showing the results of comparing the resistance to starvation stress of female transgenic flies overexpressing the Art1 gene.
FIG. 3E is a graph showing the results of comparing the resistance to oxidative stress of the female transgenic flies deficient in Art1 gene. FIG.
FIG. 3f is a graph showing the results of comparing resistance to oxidative stress of the female transgenic flies overexpressing Art1 gene.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example  One: Art1  The gene Deficient  Production and Validation of Transgenic Drosophila

(1993) Development, 118: 401-415) to induce Art1 gene deletion by receiving Art1 RNAi Drosophila line lacking Art1 gene from Bloomington Drosophila Stock Center, And an inducible gene expression system (Gene Switch).

Specifically, the Art1 RNAi Drosophila line lacking the Art1 gene was crossed with the Drosophila into which da-GS-Gal4 was introduced to regulate the expression of the Art1 gene throughout the body, and the transformed Drosophila (da-GS-Gal4 / Art1 RNAi / +), and the expression of Art1 gene was induced in the adult phase using medium supplemented with RU486 (mifepristone; Sigma-Aldrich, USA) for activating the gene switch and the control medium not added. To measure the expression level of Art1 gene, RNA was extracted and cDNA was synthesized and real time PCR was performed using Art1 primer. Specifically, the cDNA was synthesized using PrimeScript RT Reagent Kit (Takara, Japan) after DNase I (Invitrogen, USA) was treated with RNAi (Takara, Japan) Quantitative real time PCR was performed using 1 μl of the cDNA as a template using the following primers and a PCR master mix of Power SYBR Green (Applied Biosystems, USA). The PCR was performed at 95 ° C for 10 minutes, and the PCR amplification was carried out at 95 ° C for 15 seconds, 55 ° C for 30 seconds, and 72 ° C for 1 minute. The PCR amplification was carried out at 40 rpm and the amount of Art1 mRNA (FIG. 1).

FIG. 1 is a graph showing the results of measuring the expression level of Art1 gene according to the treatment of RU486 with respect to male and female fruit flies. As shown in FIG. 1, it was confirmed that the expression of Art1 gene was suppressed in da-GS-Gal4 / +> UAS-Art1 RNAi / +.

Art1 forward primer: 5'-AGCGACCTTTTAAACTAGCGTGCTG-3 '(SEQ ID NO: 2)

Art1 reverse primer: 5'-CGCGGGACGTCATCTCGTCG-3 '(SEQ ID NO: 3)

At this time, the pair of rp49 primers used is as follows:

Forward primer: 5'-ATGACCATCCGCCCAGCATA-3 '(SEQ ID NO: 4)

Reverse primer: 5'-GAGAACGCAG GCGACCGTTGG-3 '(SEQ ID NO: 5)

Example  2: Art1  The gene Deficient  Life span of transgenic flies

In order to confirm the effect of Art1 gene deletion on the life span of the transgenic Drosophila produced in Example 1, the lifespan of three sets of transgenic Drosophila flies was measured. As a method for measuring the lifespan, the transformed Drosophila was grown in the medium supplemented with RU486 and the medium not supplemented, and the number of dead bodies was counted at intervals of 3 days (FIGS. 2A and 2B).

FIG. 2A is a graph showing the results of measuring the lifespan of a female transgenic Drosophila having an Art1 gene deleted, and FIG. 2B is a graph showing a result of measuring the lifespan of a male Drosophila. As shown in FIG. 2A, the average life span of the female Drosophila was decreased by the loss of the Art1 gene, but it was confirmed that the average life span of the female Drosophila did not decrease even when the Art1 gene was deleted in the Drosophila.

Example  3: Art1  Generation of transgenic flies overexpressing genes

The present inventor intends to establish a transformed Drosophila into which Art1 has been introduced for a comparative study with Example 2 above. First, in order to prepare transgenic Drosophila having pUAST-Art1 introduced, LD28808 cDNA of CG6554 gene corresponding to Art1 of Drosophila, and pUAST transition vector for Drosophila transformation were supplied from Drosophila Genomics Resource Center. Each purified DNA was digested with NotI and XhoI restriction enzymes and inserted into the expression vector pUAS-Art1, and the expression vector was microinjected into wild-type white eye flies w1118 (Bloomington Drosophila Stock Center) Transgenic flies were obtained. The adult fruit flies were crossed again with the white flies, and the flies exhibiting the red - eye phenotype were selected and crossed again. Transfected Drosophila lines with Art1 were selected through sequencing and Zino typing for hatching Drosophila. (1993) Development, 118: 401-415) and the inducibla gene expression system (U.S.A.) in order to induce the overexpression of the transfected UAS- Gene switch (Gene Switch) was used. In order to induce the expression of Art1 at the adult stage, a bloomington Drosophila Stock Center was used. In order to express the whole body, a transgenic fruit fly (da-GS- Gal4 / + > UAS-Art1 / +).

Example  4: Art1  Effect of gene overexpression on stress resistance

From the results of Example 2, it was confirmed that the lifespan of the female Drosophila that lacked the Art1 gene was reduced. It was expected that the decrease in the lifespan was caused by a decrease in the resistance to external stress. Therefore, And to ascertain whether the overexpressed female Drosophila has increased resistance to external stress.

Example  4-1: High temperature stress resistance

(Da-GS-Gal4 / + > UAS-Art1 RNAi / +), which was produced in Example 1 and lacked the Art1 gene, and a female transgenic flora (GAS-Gal4 / +> UAS-Art1 / +) were cultivated for 10 days in a medium containing RU486, and 5 sets of 20 rabbits were placed in a high temperature incubator at 40 캜. , And the results were compared (Figs. 3A and 3B).

FIG. 3A is a graph showing the results of comparing resistance to high temperature stress of female transgenic flies deficient in Art1 gene, and FIG. 3B is a graph comparing resistance of female transgenic flies overexpressing Art1 gene to high temperature stress FIG. As shown in FIGS. 3A and 3B, it was confirmed that the resistance to high temperature stress was increased regardless of the deficiency or overexpression of Art1 gene.

Example  4-2: Kia stress resistance

(Da-GS-Gal4 / + > UAS-Art1 RNAi / +), which was produced in Example 1 and lacked the Art1 gene, and a female transgenic flora (GAS-Gal4 / + > UAS-Art1 / +) were cultured for 10 days in a medium containing RU486 and survival rate was measured every 3 hours while growing on a 1% agar medium without food 3c and 3d).

FIG. 3C is a graph showing the results of comparing the resistance against starvation of female transgenic Drosophila melanogens deficient in Art1 gene, FIG. 3D shows the results of comparing resistance to starvation stress of female Drosophila melanogaster overexpressing Art1 gene FIG. As shown in FIGS. 3c and 3d, resistance to hunger stress was increased regardless of the deficiency or overexpression of Art1 gene.

Example  4-3: Oxidative Stress Resistance

(Da-GS-Gal4 / + > UAS-Art1 RNAi / +), which was produced in Example 1 and lacked the Art1 gene, and a female transgenic flora (GAS-Gal4 / + > UAS-Art1 / +) were grown in medium containing RU486 for 10 days and fasted for 6 hours in 1% agar medium. The survival rate of Drosophila was measured every 3 hours while growing on a general medium containing paraquat (methyl viologen; Sigma-Aldrich, USA) and the results were compared (Figs. 3e and 3f).

FIG. 3E is a graph showing the results of comparing the resistance to oxidative stress of the female transgenic Drosophila with the Art1 gene deleted, and FIG. 3f shows the results of comparing the resistance to the oxidative stress of the female Drosophila with the Art1 gene overexpressed FIG. As shown in FIGS. 3E and 3F, it was confirmed that resistance to oxidative stress was reduced in the female transgenic flies deficient in Art1 gene, while resistance to oxidative stress in female transgenic flies overexpressing Art1 gene was increased.

Therefore, it is analyzed that the cause of the decrease in the life span of female transgenic flies deficient in Art1 gene is the decrease in resistance to oxidative stress.

<110> Korea University Research and Business Foundation <120> Method for regulating oxidative stress resistance of Drosophila <130> KPA151112-KR <160> 5 <170> Kopatentin 2.0 <210> 1 <211> 1746 <212> DNA <213> Artificial Sequence <220> <223> Recombinant Art 1 <400> 1 gggcagtgtt gataagcgac cttttaaact agcgtgctgt agaaacagtc ccatcattta 60 gtcgccaatt tttcgacgtg aaaaaccaaa gagacgccga agaaattaga aaatggtaag 120 ttttggtcaa aatgtgacgg caatcgacag gccagtagct gacgaacccg attcgcaggc 180 cgcacagac attcccatgg aggcagctgt ggaatcggcg accggcatca cgcccaattc 240 aaatgcgaac tcaaacaatg tggcgaagaa gctgcccgcg gagggctcca ccggtgataa 300 tcccaatgcc aacgccgacg agatgacgtc ccgcgactat tactttgact cctatgccca 360 tttcgggatc cacgaggaga tgctcaagga cgaagtgcgc accgtcacat accgcaacgc 420 catgtaccac aacaagcatc tgtttcaggg caaggtaggg ttaacttccg gggtttcccc 480 atttttcgta gcccttctca ccggggtatc tcctctttgc agaccgtact ggacgtgggt 540 tgcggtaccg gcattctctc catgttcgct gccaaagccg gtgctgctca ggtgattgcc 600 gtcgactgct ccaacatcat tgagtttgcc cgccaagtgg tcattgacaa caatctgcag 660 gatgtaatca cggttgtcaa gggcaagatt gaggagattg agcttccaaa tggtattgag 720 ggagtggaca ttattatttc cgagtggatg gggtaggttt tcatttgtag ttaagctttt 780 aaaaagtggt catatttgat ttagcatatt aaccatttta atttctttcc ttaatttagt 840 tactgcctgt tctacgaatc catgctggat acagtgttgt acgctcggga caagtggctg 900 aagaaggacg gcatgatgtt cccagatagg ggcacgctgt atatcacggc catcgaggac 960 agacagtaga aggacgagaa gatcaactgg tgggacgatg tctacggctt cgacatgagc 1020 tgcattcgta aggtggccgt gaccgagcca ctggtcgacg tggtcgatcc caagcaagtg 1080 gtatccacat cttgcatggt caaggaggtg gatctgtata ccgtgcaaaa ggccgatctt 1140 aacttctcgt ccaagttcag cctatgcatc aagcgcaacg acttcgtgca ggcattagtc 1200 acctacttta atatagagtt caccaagtgc cacaagcgtc ttgggttcag tacgtcacca 1260 gactccacat acacgcattg gaagcaaacc gtgttctacc tcgatgacca catgacggca 1320 aagaagaacg aggagatcac tggcacgttc cagatgaagc ccaacgagcg gaacaaccgt 1380 gacctggact tcgtcatcga cattaatttc aagggcgaat tgtctcaaat tcaggagtcg 1440 aacacatacc gcatgcgcta ggggcagcca taaccaatcc gatctgccat ttgggatggg 1500 gctcagagcc ctgcagcgag gatacataca atgactagac gtaaggttta ttccagtgcg 1560 aaatgtgcaa cgaattcgtg ttgcgtcttc tcctaagttt aaagtgtacc tttgttagta 1620 tacataatat tttacactcg tgcgcctgtt taaattatta ttcgaacata cttctaagca 1680 agttgtacag cagcacgtca attctcacat aataaacagt gcactaagca cgtccccttt 1740 tgctgg 1746 <210> 2 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 agcgaccttt taaactagcg tgctg 25 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 cgcgggacgt catctcgtcg 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 atgaccatcc gcccagcata 20 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 5 gagaacgcag gcgaccgttg g 21

Claims (10)

A method for controlling the oxidative stress resistance of a fruit fly comprising the step of regulating the level of expression of the Art1 gene in a fruit fly.
The method according to claim 1,
Wherein said oxidative stress resistance regulation is indicative of a sex specificity
3. The method of claim 2,
Wherein said sex-specificity is female specificity.
The method according to claim 1,
Wherein the Art1 gene consists of the nucleotide sequence of SEQ ID NO: 1.
The method according to claim 1,
The expression level of the Art1 gene may be regulated by damaging the Art1 gene itself by deletion, substitution, or insertion, inactivating the transcription factor of the Art1 gene, or by inactivating the mRNA of the Art1 gene by using siRNA or miRNA, RTI ID = 0.0 &gt; expression level. &Lt; / RTI &gt;
The method according to claim 1,
Wherein the expression level of the Art1 gene is increased by introducing an expression vector containing the Art1 gene into the embryo of a fruit fly to increase the expression level of the Art1 gene.
The method according to claim 1,
Wherein the decrease in expression level of the Art1 gene decreases the lifespan of the fruit fly.
The method according to claim 1,
Wherein an increase in the expression level of the Art1 gene increases the resistance of the fruit fly to oxidative stress.
Transgenic Drosophila having the Art1 gene introduced and overexpressed and increased resistance to oxidative stress.
(a) administering an oxidative stress-resistant candidate substance to a transformed Drosophila having the Art1 gene inactivated;
(b) measuring the survival rate of each fruit fly over time by adding oxidative stress to the transformed Drosophila in which the Art1 gene is inactivated and the transformed Drosophila in which the Art1 gene is overexpressed; And
(c) screening a candidate substance administered to the transformed Drosophila whose Art1 gene is inactivated, which shows a higher survival rate than the transgenic Drosophila that overexpressed the Art1 gene, as an oxidative stress resistant agent Way.

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