KR101602391B1 - Seawater temperature change and ocean acidification responsive genes in Scleronephthya gracillimum and method for diagnosing marine ecosystem using the same - Google Patents

Abstract

The present invention relates to a process for the production of a pink sea marlin ( Scleronephthya < RTI ID = 0.0 > gracillimum- derived gene markers, biosensors for detecting seawater temperature rise and dual stress of ocean acidification, and detection methods.

Description

Description: TECHNICAL FIELD The present invention relates to a method for diagnosing marine echinocarcinoma of pink sea bream and a method for diagnosing marine ecosystem using the same,

The present invention relates to a process for the production of a pink sea marlin ( Scleronephthya < RTI ID = 0.0 > gracillimum- derived gene markers, biosensors for detecting seawater temperature rise and marine acidification by using the gene markers, and detection methods.

Since the Industrial Revolution, human activity has increased the atmospheric carbon dioxide concentration by more than 100 ppm, and the oceans are thus a very important carbon dioxide sink that absorbs about a quarter of the additional carbon dioxide released into the atmosphere. Carbon dioxide absorbed into the ocean will form carbonate (H ₂ CO ₃ ) in the seawater, and additional absorbed carbon dioxide increases the acidity of the seawater and the temperature of the seawater.

As the acidity (hydrogen ion concentration) of seawater increases due to oceanic acidification, the concentration of carbonate ion (CO ₃ ^2- ) decreases, so it has a great influence on marine organisms composed of carbonate skeleton. In the future, when oceanic acidification causes the carbonate ion concentration in the surface sea water to steadily decline to the carbonate unsaturated state, species using the carbonate skeleton are likely to become extinct without sustaining any further life activity.

Recently, global warming has accelerated due to the increase in greenhouse gas concentration due to human activities. The global surface temperature, which is an indicator of global warming due to the greenhouse effect, has been 0.74 ± 0.18 ℃ (1906 ~ 2005) Respectively. Sea warming is rising due to global warming, and glaciers around the world are decreasing except for polar regions. This rapid change in water temperature has a great impact on the marine ecosystem, particularly by altering the metabolism of marine organisms, resulting in disturbing offshore fisheries and marine fauna and flora species.

The amount of carbon dioxide emitted by mankind during the last year was 31.6 billion tons, about 1 billion tons higher than in 2010, and beyond the ecosystem threshold of carbon dioxide amount, the ecosystem can not make a serious change. Recently, rapid acidification has been reported in the East Sea of Korea after the Industrial Revolution. This suggests that marine acidification is also proceeding seriously in the offshore waters of Korea, suggesting that the habitat of biomass with calcium carbonate skeleton may be reduced in the future.

To cope with such changes in the environment, marine organisms exhibit the property of actively coping with environmental changes by modulating the expression level of their specific genes and changing the physiology or metabolism. Regarding the method of diagnosing the response in the organism to the external environmental stress using such gene expression changes at the molecular level, there is little research on the index of environmental change related to ocean acidification, seawater temperature rise or climate change In particular, the pink sea bream ( Scleronephthya gracillimum ) have not been reported for studies on double-stressed sea- warming and marine acidification.

Therefore, in the present situation where the risk of disturbance of ecosystem due to climate change is being raised, the genes whose expression amounts are changed according to changes in the external environment such as seawater temperature rise and ocean acidification are searched and used as biomarkers, And the need to develop advanced technologies that accurately diagnose the impacts and impacts of ecosystems are urgently needed.

It is an object of the present invention to provide a gene for pink sea bream, corresponding to seawater temperature rise and marine acidification, a composition for detecting double stress of marine acid uptake using the same, a microarray, a detection kit and a detection method.

It is an object of the present invention to provide a composition for double stress detection of sea water warming and ocean acidification, which comprises the gene of the pink sea bream, corresponding to sea water temperature rise and sea acidification.

According to an embodiment of the present invention,

A polynucleotide comprising at least one selected from the group consisting of the nucleotide sequences of SEQ ID NOS: 1 to 33; A complementary sequence comprising a nucleotide sequence complementary to the polynucleotide; A polynucleotide fragment of 20 to 200 bases consecutively located in a fragment of said polynucleotide; And at least one probe selected from the group consisting of a complement of the polynucleotide fragment, to a composition for detecting double stress in sea water warming and ocean acidification.

Specifically, the probe pink sea cockscomb (Scleronephthya gracillimum . < / RTI >

In the present invention, a "probe" may be a polynucleotide, a complement of the polynucleotide, a fragment of the polynucleotide, and a complement to a fragment of a polynucleotide. Also, it may be DNA, RNA, cDNA or mRNA, which is immobilized on a microarray and hybridized with homologous DNA contained in the sample, and may be an oligomer in the case of DNA. The probe may have a repetitive sequence content ratio of at most 85% in the nucleotide sequence, preferably at most 70%, more preferably at most 50%, most preferably at most 40%. The size of the probe may be 20 to 200 bp, preferably 20 to 100 bp or 20 to 60 bp for the oligomer, 30 bp to 150 bp for the cDNA or RNA, And it is possible to select an appropriate type according to the purpose of the microarray.

More specifically, the probe of the present invention comprises a gene encoding a gene of the present invention, which expresses a change in expression level in pink sea bream and unexposed pink sea bream exposed to sea water temperature rise and ocean acidification, a fragment of the gene, The nucleic acid may be a fragment of a nucleic acid and the nucleic acid may be DNA or RNA.

The " pink sea bream " ( Scleronephthya gracillimum 'is a kind of aqueduct which is widely distributed in the coastal area of Jeju, especially in the southern coast of Seogwipo. It is distributed from southern Japan to the East China Sea. The colony (body) is very stretchy, so it is a shrub when it unfolds but turns into a lumpy shape when it contracts. There are horizontal wrinkles on the surface of the lesion, and polyp, which is highly contractive to the tube, is gathered in a lump shape. The body is light yellow, reddish yellow, orange or pink, and the polyp lumps are pink or orange. They live on rock beds with vertical walls or slopes, and they are distributed around 3 ~ 4 m to 15 m. It is a creature that makes a great contribution to the marine biodiversity of our country by providing habitat of other creatures. In particular, in the present invention, the microarray analysis of the pink sea bream of Example 3 showed that the expression level of the gene was changed with respect to the temperature rise and acidification double stress, and it was utilized as a marine index of sea water temperature change and sea water acidification .

In the present invention, the 'sea water temperature rise' may mean that the sea water temperature is higher than the average temperature, specifically, the summer maximum water temperature may be 26 ° C or more. More specifically, it may be 28 占 폚 or higher and 30 占 폚 or lower. In the case of a seawater temperature of less than 28 ° C, there is a problem that the difference in the expression change due to the temperature change is insignificant in the detection using the composition of the present invention, There is a problem that the detection rate is difficult because the survival rate of mandible is drastically lowered.

In addition, 'ocean acidification' refers to all the effects that are caused by a decrease in the average pH value of the ocean and a decrease in pH, and the average sea surface pH is about 8.2. In the present invention, The average pH of the ocean may be less than 7.8. More specifically, it may be pH 7.5 or more and pH 7.8 or less. If the acidity of seawater is less than pH 7.5, the acidity of seawater is lower than that of 26 ℃, the survival rate of pink sea mandra is lowered, and when pH is higher than 7.8, the composition of the present invention There is a problem in that the difference in the expression change due to the degree of acidification is insignificant in the detection using the probe, resulting in a lower clarity of interpretation of the result.

The present invention can be said to detect seawater or oceans in which sea water temperature is elevated and acidification occurs, particularly using a gene that responds to seawater temperature rise and marine acidification double stress. As sea water temperature rise or carbon dioxide in the atmosphere melts into seawater, ocean acidification may occur or sea water temperature rise and ocean acidification may occur at the same time. Conventionally, only sea water temperature rise has been detected to determine whether ocean acidification occurs There is a problem in that it can not be detected or the change of marine acidity is detected and only marine acidification is detected. However, one embodiment of the present invention is to detect whether double elevation of sea water temperature and ocean acidification occurs using a gene that responds to a temperature increase and an acidity increase double stress. In particular, in the case of double stress, a gene completely different from a sea water temperature rise or marine acidification single stress is expressed or remarkably lowered compared to the amount expressed in the absence of sea water warming and / or marine acidification, 5 times or more, 10 times or 15 times to 100 times, significantly less than -5 times, -10 times, or -15 times to -100 times, and the sea water temperature and acidity are slightly changed Even if it is changed, it has an advantage that it can be detected.

Another embodiment of the present invention is a polynucleotide comprising at least one polynucleotide selected from the group consisting of nucleotide sequences of SEQ ID NOS: 1 to 33; A complementary sequence comprising a nucleotide sequence complementary to the polynucleotide; A polynucleotide fragment of 20 to 200 bases consecutively located in a fragment of said polynucleotide; And at least one probe selected from the group consisting of a complement of the polynucleotide fragment, and a microarray for dual stress detection of sea water warming and ocean acidification. The genes of SEQ ID NOS: 1 to 33 are as shown in Table 1 below.

order
number Gene One dermatopontin (derm gene) 2 Glutathione S-transferase 3 spermatogenesis associated 17-like protein 4 snRNA-associated Sm-like protein 5 Olfactomedin and related extracellular matrix glycoproteins 6 TRPC channel interacting protein (enkur) 7 sll1483 precursor mRNA 8 SNF8 mRNA Css39.8 (clona9 gene) 9 Transcription elongation factor 1 homolog mRNA 10 Dynein light chain 11 ATP-dependent Clp-type protease (AAA + ATPase superfamily) 12 GDP dissociation inhibitor 13 Phosphoserine aminotransferase 14 methyltransferase 15 Glycosyl transferase, family 8 - glycogenin 16 G protein-coupled receptor 17 Ca2 + / Mg2 + -permeable cation channels (LTRPC family) 18 Collagens type IV and related proteins 19 Guanidinoacetate N-methyltransferase mRNA 20 E3 ubiquitin ligase 21 relaxin 3c precursor (Rln3c) mRNA 22 Triglyceride lipase-cholesterol esterase 23 HMG box-containing protein 24 Splicing factor, arginine / serine-rich 3 mRNA 25 Response gene to complement 32 protein 26 Retinol-binding protein I 27 DNA-binding protein jumonji / RBP2 / SMCY, contains JmjC domain 28 Vacuolar-sorting protein SNF8 mRNA (clona9 gene) 29 LIM domain related protein 30 Neurexin IV 31 Protein related to short-chain alcohol dehydrogenases 32 acyltransferase 33 Low-density lipoprotein receptors containing Ca2 + -binding EGF-like domains

The probes may be fixed by a method used in a conventional DNA microarray manufacturing method. For example, the probes may be fixed by a photolithography method, a piezoelectric printing method, a micropipetting method, or a spotting method. However, the present invention is not limited thereto, and this method is a conventional method in the production of a genomic DNA microarray, so that it can be easily performed by a person skilled in the art to which the present invention belongs.

The probe can be fixed at 2 to 100 pg per spot on the microarray, and one probe having the same base sequence per spot can be fixed, or two or more probes can be mixed and fixed. Also, the spot on the microarray can be more than one multiples, and preferably two to three multiples. The spot may be circular and may have a diameter of 50 to 500 micrometers and an interval between spots of 10 to 500 um. However, it is desirable to appropriately adjust the spot size and density according to the resolution of the microarray analysis system.

The microarray of the present invention is placed in a well of a microwell plate, for example, a 96 well plate, and the sample basin, labeling, hybridization, and washing steps are mechanically performed with automatic equipment (for example, Biomek, Genetix robo) Can be processed.

Another embodiment of the present invention is a nucleic acid construct comprising a microarray, wherein a probe immobilized on the microarray is immobilized with a nucleic acid labeled with a detectable first labeling substance and a standard nucleic acid labeled with a detectable second labeling substance And a hybridization ratio of each of the test nucleic acid and the standard nucleic acid is measured and the result is used to detect whether sea water temperature is rising or acidifying the sea, .

The term 'standard nucleic acid' refers to a nucleic acid of the pink sea bream which has not been exposed to sea water temperature rise and sea acidification. Preferably, the standard nucleic acid is obtained in a pink sea bream that has not been exposed to sea water warming and ocean acidification, and corresponds to a probe selected from the group consisting of SEQ ID NOS: 1-33.

The sample nucleic acid may be a nucleic acid extracted and separated from the specimen pink sea bream by a conventional method, and the detectable range may be various water quality environments in which the pink sea bream can survive, for example, seawater in the coast.

The first labeling substance and the second labeling substance may be the same or different and may be, for example, fluorescent substances, radioactive isotopes, chemiluminescent substances or enzymes. Examples include Cy3, Cy5, Alexa Fluor 350, Alexa Fluor 430, Alexa Fluor 488, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 594, Alexa Fluor 658, Cyanine-3, Cyanine- fluorescein, bodipy, Texas red, FITC (fluorescein isothiocyanate), rhodamine, d-NTP (including d-UTP), reactive dyes with amino-allyl modified dNTPs, Horseradish peroxidase, biotin, and the like.

In another embodiment of the present invention, there is provided a method for detecting a nucleic acid comprising the steps of (a) mixing probe nucleic acids labeled with a detectable first labeling substance with standard nucleic acids labeled with a detectable second labeling substance in the same ratio And (b) measuring the hybridization degree of each of the test nucleic acid and the standard nucleic acid with respect to the probe, and comparing the hybridization degree of the test nucleic acid with the standard nucleic acid. To a method for detecting double stress.

Specifically, in step (a), the nucleic acid of the specimen pink sea bream can be extracted from the specimen pink sea bream with a conventional method or a known kit, and cDNA can be prepared from the extracted RNA . The cDNA preparation can be performed using a suitable primer pair in a conventional RT-PCR method.

The extracted sample of the pink sea bream and the standard cDNA are labeled with different labels through random priming to obtain a labeled sample and a standard cDNA. For example, the sample cDNA can be labeled with Cy3 (blue) and the standard cDNA with Cy5 (red). The labeled sample and the standard genomic DNA are mixed at a weight ratio of 1: 1, and the mixture is reacted by treating the microarray, followed by washing.

The amount of DNA in the mixture to be treated in the microarray may be from 200 ng to 2 ug, and the mixture may be suspended and treated in the hybridization buffer. Hybridization buffers include known solutions.

In the step (b), the hybridization-completed microarray is scanned by inserting it into a detectable scanner of a labeling substance, and analyzed by hybridizing the probe information and the spot position on the microarray using the input program can do.

In the microarray, the detection kit and the detection method of the present invention, the double stress range of the sea water temperature rise and ocean acidification is the same as described above, and the probe and the sample nucleic acid may be derived from the pink sea mandrel.

The present invention relates to a biosensor capable of ascertaining whether the increase in seawater temperature and the change in acidity of seawater can be changed by measuring the change in the expression level of a specific gene derived from the pink sea mandrake corresponding to the double stress of sea- And kits.

FIG. 1 shows the results of microarray experiments using RNA extracted from a control group of pH 8.0 and a temperature-acidified double-test group (28 ° C. and pH 7.5, 28 ° C. and pH 7.8) .

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

Example  1. Pink sea mandrae ( Scleronephthya gracillimum ) Culture

The present inventors sampled the pink seabream from Seogwipo coast and cultured it in the laboratory.

Specifically, pink sea bream was collected from Seogwipo coast and cultured in natural seawater through three kinds of filters of 100, 10 and 1 ㎛. At this time, the water temperature was fixed at 24 ° C using an underwater heater, and the water temperature was adjusted to 14:10.

Example  2. Seawater temperature  Exposure to rising and acidified sea water

The pink sea bream, cultured by the method of Example 1 above, was exposed to sea water warming and acidified sea water.

Specifically, the pink sea bream was divided into two groups and each group was exposed to double stress at 28 ° C, pH 7.5, and double stress at 28 ° C, pH 7.8, for 24 hours.

The control group was cultured in natural sea water (pH 8.0, 24 ° C) for 24 hours.

Example  3. Seawater temperature  By ascending and acidification Pink sea bream  Gene change measurement

3-1. RNA Extraction of

RNA was isolated according to the method developed by the present inventors in order to measure the gene change of the pink sea bream, which was exposed to the acidified sea water by the rise of seawater temperature according to Example 2 above.

Specifically, the pink sea bream tissue of the experimental group and the control group was made into powder by using liquid nitrogen in a mortar and dissolved in a lysis solution [35 mM EDTA, 0.7 M LiCl, 7% SDS, 200 mM Tris-Cl 9.0)] was added and homogenized. The same amount of phenol solution was added to the homogenized sample, mixed well, and centrifuged for 10 minutes. The supernatant was taken to a new tube, and 8M lithium chloride (LiCl) 1/3 of the total volume was added thereto And allowed to stand at 4 DEG C for 2 hours or more. The supernatant was removed by centrifugation for 30 minutes, and the precipitate was dissolved in 300 μl of DEPC-treated water. Isopropanol was added in the same amount as 1/10 volume of 3M sodium acetate (pH 5.2) After centrifugation for about 30 minutes, the supernatant was removed and the precipitate was taken. 50 μl of a 70% ethanol solution was added to the precipitate, followed by centrifugation for 5 minutes. Then, the ethanol solution was removed, the precipitated RNA was dried, and the dried RNA was dissolved in an appropriate amount of DEPC-treated water.

3-2. cDNA Synthesis of

CDNA was synthesized from the pink sea mandrel RNA extracted by the method of 3-1 above. Specifically, Tri-reagent (Molecular Research Center Inc., USA) was used for the isolation of specific genes corresponding to acidified seawater. CDNAs of control and experimental groups were synthesized using dT-ACP1 as a template as a template and MMLV RT-ase as a reaction enzyme.

3-3. Microarray  analysis

The present inventors prepared cDNA microarrays synthesized in the above 3-2, hybridized them and scanned them.

Specifically, a sample of the pink sea bream was labeled with a fluorescent substance, purified using a PCR purification kit (Qiagen, Germany) and eluted with distilled water. The purified fluorescent label-cDNA sample was added to a hybridization buffer (3x SSC, 0.3% SDS, 50% formamide, 20 Cot Cot-1 DNA, 20 ye yeast tRNA) 30 < / RTI > to make a hybridization mixture. The hybridization mixture was denatured by heating at 95 DEG C for 3 minutes and centrifuged at 12,000 g for 30 seconds to lower the temperature of the heated hybridization compound. The prepared pink sea mandrel cDNA microarray was covered with a coverslip and the denatured hybridization mixture was pipetted. The microarray was then placed in a GT-Hyb chamber and reacted at 65 ° C for 16 hours. After hybridization, the microarray was removed from the chamber and cleaned. The microarray was rotated, dried, and stored in a dark room until scanning. The pink sea mandrel microarray, which had been tested, was scanned using an Axon GenePix 4000B scanner (Axon Instrument, USA). In the GenePix Pro 6.0 program, each point from the scanned image is graded using a gridding file and quantified to include the analysis of the Cy5 / Cy3 intensity and ratio of each point. I got a GPR file (GPR file).

From the GPR file obtained by the Jean Fix program, the following analysis was performed using the analysis program Jin Spring 7.3.1 (GeneSpring 7.3.1, Agilent Technologies, USA). The normalization was performed using LOWESS (locally weighted regression scatterplot smoothing), and the reliable gene was obtained by calculating the sum of the median values lower than the background value or the standard deviation of each pixel value Significant genes were obtained by flagging out spots that were not significant. Significant genes were also selected to show a difference of more than two times the normalized ratio value.

As a result, as shown in Table 2 and Table 3, 16 genes with significantly increased expression levels 2 times or more in the temperature raising and acidification test groups (28 ° C and pH 7.5, 28 ° C and pH 7.8) Table 2) and 17 kinds of reduced genes (Table 3) were discovered, and a total of 33 genes were shown as SEQ ID NOS: 1-33. In addition, as shown in Fig. 1, common genes among the above genes differentially expressed in the test group 2 times or more were grouped (Fig. 1).

Gene Fold change One dermatopontin (derm gene) 267.88 2 Glutathione S-transferase 59.00 3 spermatogenesis associated 17-like protein 54.17 4 snRNA-associated Sm-like protein 48.92 5 Olfactomedin and related extracellular matrix glycoproteins 46.12 6 TRPC channel interacting protein (enkur) 33.05 7 sll1483 precursor mRNA 29.98 8 SNF8 mRNA Css39.8 (clona9 gene) 28.12 9 Transcription elongation factor 1 homolog mRNA 27.48 10 Dynein light chain 27.08 11 ATP-dependent Clp-type protease (AAA + ATPase superfamily) 24.19 12 GDP dissociation inhibitor 22.84 13 Phosphoserine aminotransferase 19.15 14 methyltransferase 18.30 15 Glycosyl transferase, family 8-glycogenin 16.85 16 G protein-coupled receptor 15.21

Gene Fold change One Ca2 + / Mg2 + -permeable cation channels (LTRPC family) -100.00 2 Collagens type IV and related proteins -100.00 3 Guanidinoacetate N-methyltransferase mRNA -100.00 4 E3 ubiquitin ligase -100.00 5 relaxin 3c precursor (Rln3c) mRNA -100.00 6 Triglyceride lipase-cholesterol esterase -100.00 7 HMG box-containing protein -94.42 8 Splicing factor, arginine / serine-rich 3 mRNA -70.11 9 Response gene to complement 32 protein -69.40 10 Retinol-binding protein I -67.49 11 DNA-binding protein jumonji / RBP2 / SMCY, contains JmjC domain -46.13 12 Vacuolar-sorting protein SNF8 mRNA (clona9 gene) -45.00 13 LIM domain related protein -44.05 14 Neurexin IV -38.32 15 Protein related to short-chain alcohol dehydrogenases -37.12 16 acyltransferase -36.26 17 Low-density lipoprotein receptors containing Ca2 + -binding EGF-like domains -35.00

<110> Korea Institute of Ocean Science and Technology <120> Seawater temperature change and ocean acidification responsive          genes in Scleronephthya gracillimum and method for diagnosing          marine ecosystem using the same <130> DPP20140373KR <160> 33 <170> Kopatentin 1.71 <210> 1 <211> 775 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene <222> (1). (775) <223> dermatopontin (derm gene) <400> 1 aagcagtggt atcaacgcag agtggccatt acggccgggg aatcattcgc atcttgcccg 60 gaacaaagaa taagctcaga gtgaattttg actttttgtt aaaagatacc agaagaattt 120 ttttcagtac acgacaatga agatcatttt agtgcaaatc ctgttggtat cgttgtttgt 180 tcttgctcaa acaatgagca tttgtgcttc gacacaattt ggcgcaaaga tcgatgaaca 240 atgtccacct aatcattata tgtatcgtgt gcgaagctct ttcagcagat atcgtaagga 300 ccgttcgtgg tctatcaatt gtcgatatag tcgttccgtg actacgtcat gtcatgacag 360 tgggtatgtg aatgaatacg acggagaaat gttgtaccaa tgtccacttg gagttatcgc 420 aggttttaaa gcgatataca agaaaaataa tcacgaccga agatttcagt acaggtgttg 480 ctacaccaag tataaagtcc accgatactg tacttggtct agttataaga acgctctgga 540 aggatatttg caattcagta gtggtaacta tgctgtggtt ggagtattta gcaagcacta 600 taacgaaaaa ggggacagga tctggaaatt cttgaaatgt tcttttcaat aaatggggaa 660 ttgttgacga aataatgcaa ctattcattt cagttataat tatacgttat tttaatacgt 720 actttatgta acaatttttg gcttttttcg cacaataaaa atatcattca aagac 775 <210> 2 <211> 590 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (590) <223> Glutathione S-transferase <400> 2 ctaagttgtt acgttttgaa aagtatcttg gtgacaaacc ataccttgct ggtgacaagt 60 tgaccttccc tgattttatg ctgtatgaca tgatcgacca acataaaatc tttgatgaga 120 agttgctgga gccattcgag aagctgttgg cttatgctga tagaatcgag gttattccac 180 aaattgctgc ttacaagaaa tcagacaagt tccttgagcg tcctataaat cacccaattg 240 cttcatttac atagactctt ctgctaatgg aaagtcagtc atattgcttg gtaatcagaa 300 ttgttcaact aataggacca atttgtatgc ttgtgtatta gtgcataaaa atcattaatt 360 gtgacgtcgt gtgcaaacga agaattagtt aaaaagtttt attataacaa tattatataa 420 tgtattatga actcttagaa gaaaggcttc gttacccttc actggattgg tatcggagcg 480 ggtcaaattt cgttttgtgc atgaaaactg cttagtacca cgtggtgcaa gctaaatgcg 540 cattcagtta ttgacacata tcccgcagat gtaatcttaa aaggttttat 590 <210> 3 <211> 1214 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) <223> spermatogenesis associated 17-like protein <400> 3 aagcagtggt atcaacgcag agtggccatt acggccgggg taaacagata tggcggcctt 60 ggttaagttg ttgggggaaa aggataaaat tgtggacgaa gtatatgtca gaacaaatga 120 ggctgaaaag gcgagagagc gagaattttg ggcagtggtt aaagttcaag cttggtttcg 180 agcgcaaaga atccgaacat atttaaaatt tcttcacgac tgtgcggtgc tggtccaaag 240 acgatggaga ggtttcttgg gacgtcggaa atacagagaa aaattaaagg agcaagtttt 300 tgctatgaaa gtgaagcatt acaacgattt ggcaacaaag gtacagaaag tgtggagagg 360 tttctacacg agaaaataca tcttcaacta ttacagcaga aaacgttacc tgaaaggact 420 tgaaattaaa aatgagatta ccagatctga attggaagat ttcgaagagc agcagcaaca 480 gagacgggaa atgatcgccg aggagaagga cagaaaactt ttggaacagt gggccagaaa 540 acatcaccac ctcatcagta caaatgttca accaggcatt tataattccc ccttccaacc 600 gtatcccgat gagaaggaat attacctaaa gaattttaaa ccgtgcatgc caccaaagaa 660 ggaaaagagc aaattttacg atcccacttg caagcgttac gaccacccta ccagagatcc 720 cttacctcca atatgtccta agcctcaggg accattcagg gattctaggg cagtgcaaag 780 acagagatac aaaccgttta aaccaaccct cagagtagaa acatcttacc aatcactcga 840 agatgcacgc caaaaaaatg aaagatgaag agtgggtcac acgactcaac gatgatgttt 900 tccaaccgtt tacgagaaaa tcatatccat ttgaaccaac gttacaccag tcaacgcagt 960 ttgggcacat accatacgga aacaaattct tcagacacga atacatagat aaatttgtca 1020 taccacagaa tttcaagacc gtcgttcctc cgataccaat tttcgacaag ttaaacggta 1080 cttattccca aggtgaagtc tgaggtgaag tctcatagta tattgatttt ccaaaactaa 1140 attttaagtt gaaaacaaca tatgaactga taacttcttt atttcttgtt ccaagaaaaa 1200 cataaatcgt gtta 1214 <210> 4 <211> 394 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (394) &Lt; 223 > The snRNA-associated Sm-like protein <400> 4 aagcagtggt atcaacgcag agtggccatt acggccgggg agattgccca acagacaact 60 gataagagaa gaagagaatt catagcagaa tcgaccaaga agtttattat aagtcaaaga 120 attttcgaaa gagtttgcaa tgaatatgaa gatcgttcaa gtttttgtgc tggtattggt 180 attttctata atatactcgg gtgaaacatc tagtttgggt gaaccaaatg gtcattatgt 240 agctaaacca gcggttgctc cgtttggctg ctcctttcat cctaattggg tctattatta 300 ctgttgggga tcttgcggcc ccgatggtac agactggtgt tggattaacg tccattgtgg 360 tttcgatgcc aatgtctgcc aaaatttgga atct 394 <210> 5 <211> 460 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene <222> (1). (460) <223> Olfactomedin and related extracellular matrix glycoproteins <400> 5 aatttttgca gttattttaa tgaaaaacac agcatacatt ctaatgagga ccaaaacatg 60 tggagtaaag aacactatac acgtctatac atgtaggcct atactataac ttatatactg 120 agtggtgaac aaaatgaaat acccattttc tagatcgcta tgcatgtcta agatactacg 180 gattatggaa gattgcatct aataggtttt gttaattgaa aaaatgttcg tttcgtccat 240 tcttttctt ttgggctgtt tcagtttgct tcagtcttcg aattccagtg ggtaggttat 300 ttggcgattt ctgtcccagg cgtacaggac cttctcgcgt ggattgtaag taaccatgct 360 gttgtaacca aacttgttga caaacggaat gtctggattg atcgcagatt tggtagttgt 420 atcataggcg tagttaattt ttgtattggc tgtactgtag 460 <210> 6 <211> 1076 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene <222> (1). (1076) <223> TRPC channel interacting protein (enkur) <400> 6 aagcagtggt atcaacgcag agtggccatt acggccgggg ataggaacca agtgtttggt 60 aacaggtaaa catcttccta gagaagacaa taattataca gagaaacatg caggacgaaa 120 gcatctacaa cctgattcca agggagtatc acgctcctga aaagtcacag aggcatactt 180 ccaaatttcg tgaatcagtg agagtggagg aaaatgccca cagggcatca aacaaaacta 240 tggggcctgc taaagtgcag acgaggccac cacaggactt cctgaagaaa catgaaaatg 300 aaccaaaaat tcccgatggt tccaacttta aatacagcac ggagggtcgg aagcccccgg 360 tgcctagacg agatgaaatg cctgtacatg gttttcggac cactaagaac ttcataactc 420 aaaacgccgt tgaaaatatc atgtcagttc caaaacagcc caccaagaaa tttgcggaca 480 cgaagaaagg cgatactcaa tgtctcattc catctggcct tgagccaact tttgtccaca 540 aaaaggaatt cggtaaaacg cctcaatacc tcgaaaacag aaagaaagaa attgcacagg 600 cccaagagga atacgacatt tacgtccgag accatttcca acggggagca atgcaacagt 660 tatcacacca agaaaggagt gaaatcttag caggactgaa agccaattgg gagcaaattc 720 accatgaata tcagggtttg tcggttgtca ctgatactgc gccgaagaaa aacagaaagg 780 aaaggatgga ggcggagatg aaacaattag aaagagacat tgaaactata gaaaaacatc 840 gagtcattta catcgcaaac tgaggatgat ctgttgtttg gtaccgttat tgaatcattg 900 ttcaaaatct atttgtttta ccgaagtttg ctagtgcgta agctatgcct ttgtcctgtt 960 aacaaatgta catgcaatag agtttacatg taaagaaata ataaaatctg tgatgcccca 1020 aagaaaaaaa aaaaaaaaaa aggcgcctcg gccactctgc gttgatacca ctgctt 1076 <210> 7 <211> 421 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) <223> sll1483 precursor mRNA <400> 7 aagcagtggt atcaacgcag agtggccgag gcggcctttt tttttttttt tttttttttt 60 tgtgttaact ttcgattcta tttttcaaac taactatatt acacaactga tctagaatag 120 tttcggtgaa gcagatccat aaatctgtca aattttagtt gtcgtggcta ccaaggctcg 180 gtttacaacg gattttaaag aaaccttttg cccaagtgag gtggtttggg gtttgccaat 240 attctctttt caaacaattg aagccaatat tttggtttcc ccttctaatt ctcattttct 300 tcgagatgca aagttgacgt tagagtttgc tctgattctg gaattatccc gttctctttc 360 tcagtctcag acacgggaag tattattccg tctttttcaa tgctgttggc tcgttcactg 420 t 421 <210> 8 <211> 420 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) <223> SNF8 mRNA Css39.8 (clona9 gene) <400> 8 aagcagtggt atcaacgcag agtggccatt acggccgggg attcttcaac agctgtctga 60 ccagagtcgt acagatcagt agcctctaga ttacgctcca taacatcaca acgtttgaat 120 tgctgaatct tgatttctca caatgaagat gacaagtata gtattgtcgc tatgttcagg 180 tatcctgctt ctatcgactt tagcaaaagg tgaacctggc aggccaggga agttacctgc 240 caagggaaca agacccaaag gcaaggccaa gagccttcca ttttgtccaa tagacaagaa 300 aacatcaaaa caacttgaag caatcagggc tattttaaaa gagagaacac ctattaaatc 360 agctattgca caacaagaaa agactagtgc caggagggtt gctcgtgatg ctaagagttg 420                                                                          420 <210> 9 <211> 447 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (447) <223> Transcription elongation factor 1 homolog mRNA <400> 9 tttcttgcat tttcgtgaga atttgtaaag acttcaggtc gttctctagt tttgcgatct 60 tctttgcctg gtctgctact ttcaagatac attctgatgc aggatttgat tccgagcctc 120 cttgacacga gaattgactc ataatgtttg actgagcatg gacaagattt gtcaaaattt 180 ctacttgttt ttccaatttt tcaaccttgc tatcaggtga cgagccccga gcctcggcga 240 taacaagcgc aattaccaaa aatacccaaa acatctcgac tcgattcttc tctatattgt 300 ctgagtgtct tcacaagtta agatcgttta gttaagacgt ccaatgacga aatggtataa 360 tctgtagtgt tactgatttc cctcagttgc ctcgagttta tatatcctcc ccggccgtaa 420 tggcactctg cgttgatacc actgctt 447 <210> 10 <211> 929 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) <223> Dynein light chain <400> 10 atccgaaaac ttcacatcat atttatttgt ttttgaagaa atttgcagta acattacgcc 60 atggatgact ttcaaagtgg agaagagact gcgttcgtag tagatgaagt gagcaacatt 120 atcaaagagt caatagaagg aacgataggc ggtaacgctt accagcacaa caaagtgaac 180 caatggactt ccaacgtcgt agaacaatgt ctcaatcaac taaccaaact aggaaaacct 240 ttcaaataca tagtaacatg tgtgatcatg cagaaaaacg gtgcagggct ccatacagca 300 agttcttgct tctgggataa cacaacagac ggaagctgca cggtgaggtg ggagaacaag 360 acgatgtact gtatagtcag tgtctttggg cttgccatct agaaactgcg ctttgtgcac 420 cgaactaaca aaaacgtatc tcaactgaag tattggttct cccatctaga atatgcaccg 480 attttccgaa tattttttta aatatttaca caacgtacat aaaagcatgc tatgagttat 540 aaatttgtag ggaaaataat ttagttacaa aatgtcctgc ttaccctcta aaaacagcgc 600 ttacaaagaa ctgtcaaaaa cctatctcaa ctgaagtatt gttttcccat ccagaaacta 660 caccttaaca ccgatttaac aaaatagtat ctcaacttaa gtattgctat gtaaaaaatg 720 cactttttac accgaattga agatttttag tattttttag aggcagcttc ggtgccaaat 780 atgcaattga gtctcttgct gctacttgaa taatataatt tattcgttta aaaaataaac 840 tgccttataa aggggttagt aaaatagtag aaagttaata tcattttgag aaagtgcaaa 900 aagtatgttg aagggtattc catgtttga 929 <210> 11 <211> 614 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (614) ATP-dependent Clp-type protease (AAA + ATPase superfamily) <400> 11 aattcgtggg acgtttccca gctgtagtgc ccttccagtc tctaagagag gacatgttaa 60 tgcgaatctt gaccgaaccc aggaatgccc ttgtaccgca gtaccaggct ttgtttagta 120 tggacaagtg caatttggag ttcacagatg atgcgctcag agcgatagct cgtcaagcca 180 tggaacgtaa aactggtgct agaggtcttc gggcaatctt ggaaaacata ttgctggatg 240 caatgtttga tgtgcctggg tccgaaatag tcgacgtcgt tgttgaggaa gacaccgtga 300 aacaaggcaa accttcacgc tatgtcaccc gtccaaaaga agactcttgt gagacaaacg 360 aagaggcagc ttcgtataat aaaaaatcac agcacaaaat ccttgaaggg cagcttctcc 420 gtcgaacact cgtgaacttt ggtatctttt gattaatttt gaaggatatt gaggatttat 480 gaaaagagac tttctaagtg agcaattata ttgccattaa atgttaaaaa gcagcattga 540 cttttaattc gttcttgaaa tggttactat agtttgacta tgaaaagaag cattgctttt 600 tataattcat gtgg 614 <210> 12 <211> 1096 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (1096) <223> GDP dissociation inhibitor <400> 12 aagcagtggt atcaacgcag agtggccata cagccgggct tacattacaa agtcgttcaa 60 atggatctcc tgtacgcgtt attgaactga caccatcttc tttggcttgt ccagaatccc 120 tttgcgttgt tcattttact actgaaactg tagtaaatgc cttcgaagat ctcaatttgt 180 atgtagaaga tctgtttacc acaccagacg cagctgacaa cgatagtgaa aagccagtcg 240 tgttgtggtc agtttatttt aatatgaaga acggtgttcc cacaaacatt gataaagttc 300 ctgttaatgt cagaataacg tcgttacctt cagctcattt ggactttgaa gatgccatga 360 gtgaggctaa ggatatcttc acctctatat gtccaaatga agatttctta ccagctgttc 420 ctaacccaga agacataata tgggatgata ttggaagttg ttcgaaggag gaaccatgtg 480 aacaagctag tggtgaagtc attcctgcca ttgacgcacc aagttcagaa gtcctaactg 540 aagagtctgt tgaagactct tacgatgatc cattagaaat gaatgataaa gaagtgtcaa 600 actctgaaga tatatcaagt tctgagaaca gacccgagac agagattgaa aaatgaagta 660 taattgtgcg gttgcatctc aagcttttcg gcccgaaagg tttttaacct ttaattttag 720 caataagcaa tccttgtggc gaaggactat aaatgattac atgtgatttt cgtgcctctg 780 gctaaaaatg gaaaaccagt aggaaaccac tagtattaaa aaattagata aaggctgttt 840 tttatatgtc gtaatttaat ggcatagtcc gttggtaatt agtgcataat tatgatgaat 900 tgtctgtcgg aaaaaattca gcttcattga tcatggtgtg gacaatcgat cgcagaacag 960 caccgatgaa ttatgacatg tgaacaccag tggtgaacac agacaatgtt gctctgcaaa 1020 taatttagtt tatccataaa ttagtgaaga taattagtct atagaaatgt ctgtgaggca 1080 aaaattaaaa gtcatc 1096 <210> 13 <211> 1301 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) <223> Phosphoserine aminotransferase <400> 13 gtgtattctg gacatcgaat tattaccctt ctcctgatcc gaattttaag tgtatttttg 60 tctttaaaaa tagtcaatca cacgctatat atcgcaatct gggactaagt tagatgatga 120 taccaacaat taatgatgct accgaccacg gattgtctac ggattctaaa tccgttttgg 180 cgccagcaaa gcgttgacac tgcctatagg gccattaggt aatgtcgcag aatagccact 240 ggatcactcg catcagcagt acgttagata tgacataaga atacaatatt atgtggaaat 300 gctattcgat tttcgttatg tgagttatcc tctgaaagtg tttcagtacg tgaataaatt 360 attagaattt tacttaattt aaaacaattc cctcatattt ctacaatctt tacttgctgt 420 atttttcctg gaaattcttc atgaattcta tcaaatgttc aacctcatgt aacatcactg 480 cattatacag ggacactctt aaaccaccaa cagatctgtg tccgttaacg tgcttatatc 540 cctgttcgtc tgcttcagca acaaatttct tttccagatc ctcatttggt aatcgaaaca 600 caacgttcat tcttgaacga gctttgcttt cgaccagggg gttgtaaaat ccgtttgaat 660 tttccaaaaa gtggtacatt gtagaagact tctcgtcaca tcgtttcgcc atttctgtca 720 gaccgccttc ttttatgatc cattttaata caagccccat gatgtaaaca ctatatgttg 780 gcggcgtgtt aaacacagaa tttgcatcag caacaacctt ataattaaaa cttgtgggac 840 aacatggtaa gcttctgtcg agtaaatcct cacgaacaat aacaaccgtt actccagggc 900 atccgatatt cttttgagca ccagctatga tacacccaaa ctttgacacg tccacaggcc 960 tcgtcaaaaa gttcgaagac atatcacaaa ccaaaggaac gccgtttgtc tcgggtataa 1020 aatcaaattc aactccgtga actgtctcgt ttgcacagta ataaacatac gaggcgtctt 1080 tattcaactt ccagtctttc tcatcaggga ttgtggtgaa agtcttaggc tttggaaaaa 1140 cgtaattgac ctgaccatat tttgctgcct cgttagcagc cttggatgac catgttccgg 1200 tgacaagata gtctgcactg cccccctcct tcataagatt cattgggaca aatgaaaact 1260 gcccatatcc accaccttga cagaaaatga ccttgtagtt g 1301 <210> 14 <211> 306 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) <223> methyltransferase <400> 14 aaaatatcac gcattgcaaa tttttgtcag gagattggga atatgcaaaa tcattgctag 60 aacctatgtc atttgatgtt attctcactt cagagactat ttataacagt gatgtacaaa 120 aatcattgta cgaacttata aagttctcaa tgaaggcctc tggcgtggcc ttcgttgcgg 180 ctaagagtta ttattttggc gtgggtggtg gcatagacca gttttccaag ctggttaagc 240 aagatggggc atttgatatt tctagtgtta caatgagttg tgaaggagtg aaaagagaaa 300 tattag 306 <210> 15 <211> 203 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) <223> Glycosyl transferase, family 8-glycogenin <400> 15 ctccaggatg cttcttttgc tgatggaaaa atcctaaaag ctttttaagt ctagaggcca 60 ttgagttcct tccatttcgc tgagcatgaa taccatgaaa catcaaatct tgaaggcaaa 120 acatttcaat tttctcaaga tatacatctt ttgcagcgcg ccatgctttg aaatagatct 180 caccgtatac aaccagtgtt cct 203 <210> 16 <211> 1771 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (1771) <223> G protein-coupled receptor <400> 16 aagcagtggt atcaacgcag agtggccatt acggccgggg atacgaggca gggttactct 60 aagggcacac gcgcgcatca attgacttat ccgttcagtt taattcaacg taaataaagt 120 tcgtgacaca tgcagagttt ggaaaattgc acgggttgtt ccaatacaac tggcaacaac 180 acagaatcgg cggtctacaa caagcttttc tccgtggcga tattcataac catttacgtg 240 ttgttcctgc tattcgcagt cattggtaat ggccttgtat gttggattgt cctcacaaac 300 cgtagaatgc acgattcgac aaatattctt ttggttaatt tggctttaag cgatttactc 360 ttggctgtcg ccagcacatt tcaagttgcg gattttgctg tcaaggattt aaatcttggt 420 gatgttggct gtaaattcca aatcaacatg gtgaatattc cttacggtgc ttcttgtttg 480 acgatcgcct ttatttcctt cgagcgatac tatgccatat gccgacctat ggacttagat 540 tatgttaagg aacggttgaa atatatcatt ccgtcaatat ggcttctttc gtttacgata 600 tactttccaa ctttgtatta ttgtggtagt aatctgaagc gtgaaggtga tcaactcagc 660 tgtgactgca catatcgttg gccgagtctg aaagcgaaga atattcacgg tgttttgatt 720 gttgtgtttt tgtatttcat accattcacg gtcgcatcat gtttttattc tgtcgttgta 780 cgtagattga agaaggtaat ccctggggag aatcaaggaa atattgctgt gtataaatca 840 agacgcgggg ttgttagaat ggtcatggtt acgttgatcg tattttttat tgcttggaca 900 ccttacaaca ttctttactt gctgaaaaga ttggaggttg actttcgcag tgtctaccca 960 ttcgtgtggt atccagccct tttgttagct ataattcact gtatatgtaa cccgatcata 1020 tattgtctac ttggaagtaa ctttcgaaag gctttcaaga caatattatg ttgtcgtagt 1080 tgtcacgtcc ctggcttcgt tcaagatttc cttggtcaga gttccactgg gaggacaaca 1140 cgtcaggcaa cgcgtcaagt acacagcaat gacgtgcaga tagaaacttg attctaacat 1200 acagtctgta ccgaatatat ttgaggtatt tcgaatcgga agaagccatc ctatttggac 1260 gtcagttttt gagaatctgc gcatgcatgt gttgtaaaag aattactttc tacagatcca 1320 cgaaagttat ttcgtaataa aggaccgaaa caaatagagg aacaatattt tagttaaaaa 1380 acttaacgca gtagagtaga aatttgacag atgcaaccac agtgaaattt gttgaaattg 1440 ccatttaaca tgtacacttt tataccccat gttcaaacaa tattgataca gatattaaag 1500 gtaaatgtca agaattgaaa tacatcaaag ttttaaatat tttggaatta aatcatttta 1560 aattttaaag tctacatgca acagaggcca gaaaagtctc ttcttgcctt cgtcacccat 1620 cgcatttggt gccagttttt tttccagatc taactcgtca atttgttgac aagctcgcta 1680 caaagtgttg cggcataaat cgggtataat gtgatctatg tggtatgaag atgatgggtc 1740 actaatatcc accttctcca tcatgcctac c 1771 <210> 17 <211> 1138 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (1138) Ca2 + / Mg2 + -permeable cation channels (LTRPC family) <400> 17 aagcagtggt atcaacgcag agtggccatt acggccgggg gtctgaaagg gaaagacaga 60 aataattaaa acgtaaacaa caaacgcaat agatttatca cggttaacgg tgacttccaa 120 atagctgtaa aacaaggttg gttttgtgaa tatcgtgctt ttaaagtcat gggaaatagt 180 agctcacacg aaacatactc ttcttcaaac gctcacaaag acggctatca ttactgggca 240 aagtttgaga ccgagccttt tgatgagggg gcttctcgat atgcttttaa aggcacctat 300 ataggagaag gcccaatgaa tggccgtacc tgtgtcacta aggtgttcaa aaagaagtac 360 gaaaagaatt ttgacatgtg ggtgcctgat ttggcttcaa gcaagaaagc tcagatgttc 420 gcagagaagt tcaatactaa agaactttac caacttgata ttgagccgaa acgagagttg 480 aattatgtta ttcctcttat tgccaaaaca gacgcactat cacggtttta tttgctttgg 540 tttatcccat ttgggtctga ggataaaaga tatgttatgc cacaagagta tgtagctatt 600 gagccgtata tagatggaga ctatgaaaag ttcaactcta acggggggtt tgaggatcag 660 aacctaagtt cactactacc agccttctct cactggactt gggaaatcag tggtcataaa 720 tatatggtat gtgacttgca aggagtaaaa ggcgatggtg aatataagct cacggacccc 780 gtggtgcact caatagatca gattttcggc aaaaccgatt tgggtgtggt cggcatggag 840 aaagtcctgg ctaatcatga gtgtaacttc atttgtcaag agctggggct acagaatccg 900 atgaaacatg tctatttgcc agctttagca agaagtacca cctattcctt tcaactaaca 960 gaggaagaaa agttaagaaa taagatagga gtaagccgcc atttccaagt catgacggcc 1020 attctggaat aaatcatgac ttagttgcta ttagttgcta tctccatgca taccaaaaga 1080 gacttgacat gtatatttgg tgttgtttca agctaaaata ataaaagaaa ggtacttt 1138 <210> 18 <211> 1074 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) <223> Collagens type IV and related proteins <400> 18 aagcagtggt atcaacgcag agtggccatt acggccgggg actatagacc tgcacattgg 60 ttggaaccac cccaagaaga aagaagacac tcagttaatc atgaaatcga agacattcct 120 atcactggtg attttctact ggtttatacc aaagtacagt catcaagcat gcggtccagc 180 atcagctccg acattgccat cagatccaaa tgctccgatt taccagagac taaaaaagat 240 tccttcgcct aaaacaactg cccaaaagat tttggacaat tttataaagt ttactccagg 300 agtcgcaaag agacgaataa ggaccgaaga tatcattttt gtaatggatg gatcaggttc 360 agttggtcaa tgtgaatttg atgaaggaaa gaacggtttg attagtttga taaagatatg 420 ccagaaaaca ggttattctt gcgtacatgc tgggattacc tttgcctcgt ctgcacagag 480 gaactttaag ttcttaccaa gagcgcaagc aattcaaaaa atgagagcag taccatatcc 540 tggtgggtgg acaaatacag cagcaggttt ggaagaagca cgcaaacttt ttcttgacag 600 gaattcagga gggcgacgcc gagcaaaaga gatggttttt cttatgactg acggacagtc 660 gaacacaaat tcccacctta ccgtaccgaa cgcgaaaaag ctcaaagata tgggtgtgga 720 aatttttgtt gtagctgttg gtggttttag ctcaggtatt aaagaaattg ctgaagttgc 780 ttcatatcca ccagagaagc atgtttaccg aatttataaa aatggagact tcccatgggt 840 catgaaatta gttctagaga aagtgtcacg gggaaggtat aaagcgatca aaactccatc 900 ccagtgcagc taaagagatt tctacctgaa ctcatgagct ttcgtacaga taaaaagtag 960 tataacatga ataattcaca aaatatgatt aagtaatgaa ataaaaacgt agagcaatag 1020 atccagcctg catgacgact taataaattt gtattgaaga aataataaat ttat 1074 <210> 19 <211> 1249 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (1249) <223> Guanidinoacetate N-methyltransferase mRNA <400> 19 aagcagtggt atcaacgcag agtggccatt acggccgggg gccatgaatt cgatgagagc 60 aaagcgtttt aagaatatcg ttcgaatgta ggcagtacat atttaaagat tttttgctca 120 tatggaaggc gaagccgaaa gggaggaaac tctaggcaga gcagacgccg cagagcaaca 180 aggcgagagg attaattttg tgtccaggga tggccaagaa acatgcgaag gccagccatc 240 ttcgacacct tcaatgtcgt tgaattctat gcctgccaat tccgaaacac atgtggacac 300 cagtgctata actttaaatg aggagcaagc atcgcaacag tttgctgctg aatcagaaaa 360 gaaatctgta gaaaaatacg gtaatgccga ttgggttgaa ggtgcaacgt cacaaagtga 420 acgtggaaat ggtactgaac taaatgcatc cgttgctgaa aacatcatgt ctgccaacac 480 cgagacaact tcaaatcaag tgactgttat ggggaggaat attgatgatg ttcaacgagt 540 tgtggtgcaa gaaacagtta ctggtatagt tgaggtagat tctggaaaat ccaggtatgc 600 tgacgaattg tcgagagatc ctgacttttc aattgaggaa gagaaaacaa ccatttcaaa 660 gagtggaata aaagcagatc aaaacgaggt tttagacaaa tgcttttaca atcttagaaa 720 acctacaaac ggatgaagca agaaacgttg atgatgggca agttacgcag gaaattgtaa 780 agcaaccaag cgagaagaaa aagagcgaga gtaacaattt acctttagcg tttaatgtta 840 ttggcacaaa ttttgacggc aagagtggca taactgagga ggataattcg ggcttgagat 900 gtgctgctga gttgaagagt cctccgtgtg ttgaagaaat tgaatatacc atttcaaaaa 960 gtgaaaaacc gggtttgaat gaggttttag acaaatgtta taaagccgta caagagatgc 1020 aaataggtgg gtcattaaat actcataacg ttgtgcagca atcaaaagtg aacaaagaag 1080 gaaagaacaa ggcgggcgag aatacaaaga gcacaaagaa aggcgtggag aacaaaaagg 1140 ataccaagaa agacgacgaa agaaaaaagg atgcaaaaaa agatgacgag aagaaaaagg 1200 actgcagaga aagacgccaa aaaagaaaaa gaaatacaaa gaaagacgg 1249 <210> 20 <211> 1701 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (1701) <223> E3 ubiquitin ligase <400> 20 aagcagtggt atcaacgcag agtggccatt acggccgggg gactgatttg tatcatgtat 60 gagaactgaa tgcgaatttt aatttttaac ttcgaaatca gtgttggcgt gtagtaagtt 120 tgtgtggaca cgtttaatac agtattttga aaggtttttg taaacattac gtggaaggaa 180 tgaagtttat caaaaaaaag ctgaaataat tctttgttgc gaaatggaag aggcaatcac 240 ttgcaaggtt tgtgcagaaa aattttgtga cgaagaagga cttattccaa ggatactaac 300 cgcttgtggg catagtttct gtgagtcttg tctaagtaaa ctacaaaata agaagtctat 360 acgtattaaa ggtgttactc tgtatcaata tgagataaaa tgtccaaaat gtaaaacaga 420 cacgatttct gacaaacgca gtagctatct tccaaagaac tatgcttact tagacttact 480 cgatgaagtt gtaagaagtg gaagagatca tttaaaattt tgtcatgtac acccaaacta 540 tgtcctagac atgaga tgtatgtatt atgaagaaga ggaagcagtt gtacacata 600 tggcaagcat aagacacaca agatgtctaa attgtcatgg ttttcagaca agcaagcaga 660 atctatacaga tacaaaattg attgtataga tgatgtcatc agtcattata atgatcttag 720 tttatctctg gaagcaagga agcataacct taagcataaa tctgaacata ttcaagatgc 780 aataaactca agattttgcc atatacgatc agagttaaac aatatccttg atgcaaagca 840 agaatctgtg ttgagtcagt tgaataagtt ctccacttct caaatgtctt tgttgaaaat 900 gcaagaagaa gctgcaatga atttcacctc caaacttgat gaaaagagaa attgtgctaa 960 atatttcctt ggcacagctg aagaatgcag tattgcaaaa gaaaaaaaat tacatgatga 1020 catgctgaag tgccttgatg atgcacaaag tcataaaatg ttaacaaata atatgtatga 1080 ggtacagatg gataactcag atcacattgt caatagtgtt agggagatgg tcaatgattg 1140 ggtttgcaag gttgaagaaa caaagctaga accatcactg gaatctgtgt ctacattgtt 1200 ggagcaacaa acaccttctt ttggagacag aatggatgtt ctgtttggtg atgctagcga 1260 aatcaccatg aagaatttat ttgaagaaat tgattgtcaa ttatggtgtg atggtaagct 1320 gcaacattgg gaaatagact gtgatggaaa aacagagtgt ggtctagaaa cagagccatc 1380 agatgaagag tccaacatag aatgtgttgc aagtgatgag gatgactaag aatatagtta 1440 aacacgtaga attttctggt tgtttataac accaacacaa ttgttataaa gaaagctgtt 1500 aagagatgaa gagatgaacc ttcataggtt tattactttg ggctattggt tacagccccc 1560 ggctgataga caaaaagtaa tgaaaccagg ctatgtatat taagttgcca tatatttcat 1620 ttatatacct tcactttagt ttttcccatg tgcactgatt ataaatacat gattggaata 1680 gacttttttc tgaaatatag t 1701 <210> 21 <211> 348 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (348) <223> relaxin 3c precursor (Rln3c) mRNA <400> 21 aagcagtggt atcaacgcag agtggccatt acggccgggg atataacaga aaactgcggt 60 tgaatttggg tgaagaaagg cgattaaaca accgtaaaga tgttaaagat ttgtctgttt 120 gttgttgcaa ttacatgctc catatttgct gatgcaggaa ggcttgatga atactttcgg 180 aatgatgaca acgaaaacga tgtcacgttc agagaaattg aacaaacaag gcgtggcata 240 cccggacatc tcttctgcga aagaaatggt gctggttgta aactaaagac tcaagctact 300 gcttacataa aagatcaata cacggatgga gcagtattct gtcaaaaa 348 <210> 22 <211> 472 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (472) <223> Triglyceride lipase-cholesterol esterase <400> 22 tactcctaaa aaatatgatg tttccaaaat ccgggtaccc gtggcgctgt tttccgctga 60 ccaggactgg ctggcaactc ccaaagatgt ttccttacta gaagctaaac tccaaacggt 120 ggttttcaac aaaaatctta cgtcttggga ccaccttgat ttcatttggg gaatggatgc 180 caccactttg atttatgaag acataaatgt tctgctgaaa aaatatggac cacgaggaat 240 atgatcggga gtgtgaactt ggatgtcgcc ttggttttta agcttagaca ggatacttaa 300 gctattattg agaccagggg tttccacaaa gcttttgttg taacgttgct gataccttct 360 actagcgcgt atttttagtt ttttttacta actagtatcg tcttagggga ttcatggatt 420 aggttaaatg ctttttacca taactttggt gtaatttggg ttatatagtg tc 472 <210> 23 <211> 1123 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) 1123 <223> HMG box-containing protein <400> 23 aagcagtggt atcaacgcag agtggccatt acggccgggg attctttttc gaaagtaaac 60 aaagtaaaca cattttgtaa agagttggtt gtaaaaatgc tttcttttac gaggttttgt 120 acgagatctt gtggtcctat cttgcagcaa tgtgcagtga gatgcacagc tgttttacaa 180 gtgaggtatg caagagctaa agtattgcat gaccttcctg agaagccgaa aagaccgcca 240 agcacttggg tcttattcct gaaagaacgc cgtaatgaac tgcgacaaca gcccgaatac 300 gaggacttgt ctctgatgga aatgacggca aagataagcg aagaatggcg gaattttgac 360 gagcttgata agataccgta cagggaaaaa tatgacgaat ctttgggcga ttatagacgg 420 cgaatacaag agtacaacat gaatttaacg tcagatgata aacgtatttt acgcaatatt 480 aagaaagatg gaaagcgaga tatacgtgca tttgagaaag agtttccaaa gccaaaatat 540 cacggcaatg gttatatttt gtttgtgaag tcttgtcata aagaaagtcc gcgcagagag 600 ggtgaagata tgaaggactg gatccgagag tgtgctcaga aatggaaaaa tcttgatgaa 660 gaaacgaaag ataagtttaa cgagcaagcg tcgactttat tggggcaata cagagaagaa 720 atgagagaat ggaaagataa atataaaggg tggaaaaaaa gtggcgcatc tcacaactaa 780 cttacaaaag ttaaaatcta tgtccataaa aacacttcat atgtaaagga atatgatatt 840 tgtaaataga aaaaactcag ttttgttgca gttttcacat tgaactacca ttgacatgca 900 cctaaccacc ttggtttcaa tttggttgtg aaatcccaat catatcaaac aagaggtatt 960 aacctagggg ttgtagtctg tgataaaagt atccccagaa aaatatagat aaattagaac 1020 atggttcata gccctggtgg atatatatat taaaagttca gattggcata tttggagaaa 1080 ttgcatggga aagacattac catacacagc aaagattcct aaa 1123 <210> 24 <211> 877 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene <222> (1). (877) <223> Splicing factor, arginine / serine-rich 3 mRNA <400> 24 gcagtggtat caacgcagag tggccattac ggccggggac ttgatcttag cacttcaagc 60 aagaaacagt ttgaagaaga gcacaagaac gatgaagagt tttgcgctgt tttgtttcct 120 attgttggct tcggcctgcg tttttgtcgc tcgagcggag gaagaaactc ccgcagtcga 180 tgacgctccc aaaatggagg atcagcctga gacagctgac gtaacagctg acgaggagca 240 agccgatgaa gaggaggcgg aatccgatga agaaaaagag gaagaagacg aagaagagaa 300 agaagccgac gaagaaaacg aagatgaaga gaccgaagaa gatgaagaag aaacaactga 360 tgttgccgat gacgagcccg agaagcgagg ttggaaatgc aaatactaca ggaagggaag 420 gctcaggtgc aaggcctgtg gatgcagacg tgtgtgcaaa gccagatggt gcgtccgacg 480 tgtgtgcaga cgatatagat gtggtaccag gagggtgtgt cgtttggtga ccagatacag 540 aaccatcaga tacaaatact acttccctta ctaccgtgga tatgggcctc gctggatatg 600 gaagaccaaa cgagttgcat acaaagtcag acactgcgta aatgtaccca gattctgcac 660 ccggtgccgc aacgttcgtt accgatgtgg aaatatctgc aagactgtgt gcagatacag 720 aaagtgcttc ccatactacc gcggataaac ggtcacactt cgtgaaagat acctggttaa 780 attacggata attatgattg gaaaattattt actttactag cattgcatgt agaacaaagc 840 acagaattgt caagtaattt tgtgttgagg aaatata 877 <210> 25 <211> 557 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (557) <223> Response gene to complement 32 protein <400> 25 aagcagtggt atcaacgcag agtagccatt acggccgggg gagattcaag atatcttaac 60 tgaacgtcct cgcagttttg acatggaggt gagaaggtat agagaaaatc aacgagatcc 120 tctgtcaaaa acgttcatgg gaatacgact aaaatggagc gtgtggcgag aagcaatggt 180 tccacaaaat gtcggtgctg taaacccatg gctttattct cctcgtactt ctcgccaggg 240 tgttacaaca cattattaca acaagaacac tcgtcaggaa cttgcccaac aatgtcctga 300 gaacgaaaat tgttgtggtg tttacgaatt caaagttgaa agaaactatg gagaagcggt 360 tgtgtacatt ggctctacac acagacgagc agggagatct atgtatcgac ggataagcga 420 atatctcacc aacggaagcc atataagaag gatcattcag agagctcttg ataggcagtt 480 taaaatctat gtacgatgga tgaaaataag agcacccttt tatcattggg caaacgggag 540 acaactagcg gaacggc 557 <210> 26 <211> 719 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene <222> (1) (719) <223> Retinol-binding protein I <400> 26 aagcagtggt atcaacgcag agtggccatt acggccgggg agagttcctc gaccatctga 60 agagacaaaa gtcaactgca acgaaatcaa atttggtgtt tgacaattcc cccaagcaat 120 cattatgaac gttcaatttt tggtgtcatt cgttattgca ttatcgcttg ttcccggtcg 180 cgaagcggtg cgctgcagaa aaatagaaga aggcgtgtgc tgtggaaatg gtaaatgcga 240 tacatcctgt ggcgattgtg atggtggctg caatgacctg tgccagttta ctcattgtaa 300 ccccttagaa tggttgaaat gtgccggtaa agtggctgca tgtgcaggtg cctgtatacc 360 taatatcaga tctccagcgt gtattgcgtg cctcggtccg ttgtggaaca catgtaaaaa 420 gtgtttttca aaagaagtta gtgtacaagc agttttggaa gataacaaat acatgttaac 480 tgcttatcac gattatttat atgacatata tagaaagaaa tgagccatga acgaaatttc 540 accagtggca atggctcacg agaacaaaga aaaggcgaac agttaaaatg aacgttaagc 600 atttacaact ttattatttc agtagcatta atattttagt aaggaattaa tagcatgaag 660 ctaaaattct ttctttagca tgcatgttaa tcacatgatc atattaaata tattgtaac 719 <210> 27 <211> 2040 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (2040) <223> DNA-binding protein jumonji / RBP2 / SMCY, contains JmjC domain <400> 27 agccattaac agtgaatgtc ttaacgtctt cttcgggtag tgttagtagt ccgccactaa 60 tatcaccttc actaatgtca cccggattgt tatctccatc gagacattac tcgctactct 120 cgccgtccga actaaagaag accagtcaac agggtactgg ctttagcgct cttggaagtg 180 ttaaagtaag tttgctcttg gaacctggtg ccttaccgca accaccagcc ttacctcaga 240 cgccttgtcc tcgtgagaaa ttgtcgccac caacacccag cattcgtgtc gaaactaaat 300 ctgaagctta ttcgactgag ctacaaaatc tatgcctttc tccatcccaa ccaatcacag 360 ttataagagg actcgataaa gcacttggca tagatcttag tttattctca acaaaagcat 420 tgttggatgc tcatgcagac cacgaagttg aagttcgaac acagaaacag cagttgtcag 480 acgaaaatat tgacgatgaa ggacgaaagg tttggctttg tgaaagtagc agatcgtatt 540 ccactattgc taaatatgct caataccagg ctacgacctt tcaagagtct ttggagcagg 600 aagatcaaga acgtaatctg tcagacagtg attcatcgtc atctggtcag aaaaggaaga 660 aacgaggaac ccgaaaaatt ataaagtttg gtacaaactg tgatctttcc gatgaaaaga 720 aatggaaagc gcaattcctt gaacttaata aactgcctcc gtttttaaga gtcttctcac 780 ctggcaacat gcttagtcat gtcgatcacg ttatcttggg aatgaattca gttcagttat 840 acatgaagat tccaggatgt cgtactccag gtcatcaaga aaacttgaac tttagctccg 900 ttaatatcaa tattggtcct ggagattgtg agtggtttgg tgtgcctttt gaatattggg 960 gagctattca caatttttgt gagaggaata atgttgactt cttaatggga tcttggtggc 1020 caattcttga agatttgttt gaagagcggg tcccggtgta tcggttcatt cagaaacccg 1080 gtgacttagt atgggtgaat gctggtacag tgcattgggt gcaagctgtt ggatggtgta 1140 ataatgtcgc ctggaacgtt ggaccattaa cagaatttca gtacaatatg gccgtcgaaa 1200 gatacgagtg gaacaaacta caaggaaata gatctattgt tcctatgatt ttattgacgt 1260 ggaatttagc ccgaaatata aaaatttcgg aaaagaaact gtatgaacac atgaggcaag 1320 tgctggcacg agtgttgaaa tactgtcagg taactttgga ttgcttgaag gaagctggtg 1380 tgacagtgac actacaaaga cgagttgagc acgaagctgc ccattattgt tctgtatgcg 1440 agactgaagt gtttaacatt ctatttgtaa caaacaataa gaaacaccag gttcactgtc 1500 aggattgtgc aagaaagacc agcgatgttc ttcaaggatt tgttgtctta caacagtttc 1560 ccatggaaga gctgtataaa acgtacaatg acttcaggtt gtaccagcag tcttcgacaa 1620 gtgtgatcca accatagttg taagtgttgg cttgcagtgt cttagaagtc taacgatttt 1680 ttgtcgtgag aatagcgtta acgagagtat aaattacgaa cattttgtta tatagtaggg 1740 tttattttcg tagtttgctt ggtcccacca accatacatg tagatagcga ttttcagaat 1800 tgaactgacg acggtgatta catgctgatg accaactcga ggatattttg gccgaccaag 1860 attaagattc ggtcaaaatc gtcatttcca ccaatgttaa ccgtaatatt tatactactt 1920 ctcactacgg taaaccttga tatacctttc attacacaag cacatatttt acatggttaa 1980 aagcaccctt tttgattgag aaaacggaag catgacttag gtctagtaca tcacattgca 2040                                                                         2040 <210> 28 <211> 140 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (140) <223> Vacuolar-sorting protein SNF8 mRNA (clona9 gene) <400> 28 aagcagtggt atcaacgcag agtggccatt acggccgggg attcttttaa ctatcgcaac 60 caaaaacgag gctaaaaatt gcatagagaa aaaaaatatg tgaattccaa gcaaataatc 120 caaaagcaca gctttttttt 140 <210> 29 <211> 786 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (786) <223> LIM domain related protein <400> 29 aagcagtggt atcaacgcag agtggccatt acggccgggg atcaaattat ttgacactga 60 gtatatacgg ctacaacgca aggaggtgaa aacaatgtct tacaggagcg gtgccggctc 120 aacaagccgt tgtacacact gtcggcgtca aattcaacct gaagagaaga agttgaatta 180 tcaagatcaa cctttccatg ctgattgttt catctgtgaa cactgccgta atcctcttgg 240 aactgatagt tttgtgaaga gggatgataa acgctattgc caaaaatgtt ttgagcgcct 300 atttgcaaag aactgcaatg cttgtggtga catcataaag actacttccg ttgattatga 360 aggtaacgcc taccacagtg attgcttcac atgtagcgaa tgccgcaatc ctttggctgg 420 aaagaagttc cataaaatgg gaagaaagct ggtgtgtaaa gcatgttacc gtgataagta 480 cgccaaaatg tgcgattact gcaagcaagt gattgaagct aacatcaagt tcgttgtgga 540 tgatgagaaa acctatcatc gtgagtgctt cacttgcagc aagtgtggtc gacctattga 600 tggagagaaa tacaacatca agggggacaa ccgtatctgt ttgaagtgtc ctacctagga 660 tttcacaaaa ggaactacct taatgattct acacctctgt atcgaaaatt gctgtataca 720 atatctcctc gttgttagtt tatatccaga aagtagttaa aattaaaagc aatgttttca 780 atacct 786 <210> 30 <211> 481 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (481) <223> Neurexin IV <400> 30 taacagtgta ttattgaacc cgcagtacgg ttggtgggtg tcacgtcaag gttcaaagat 60 gaattactgg ggtggagcgg cagtcaacag tggaaaatgt gcatgtggaa tgaccaacag 120 ctgtgcaggg ggaagacaat gtaattgtga caagaatgat gtaaaatggc gtgaagacag 180 tggatacctg acagacaaga aaacgctgcc tgttactgga ctgagatttg gagatgctga 240 cggtactcta tattatacat ggcaaagaga gtctggttat catacactgg gaaaattgcg 300 ttgctggggt taagagatgg aacgccagaa aacgatgtga ttaatatgat ttcttcatta 360 atccaacttt gattaatttt agtttagatt tgtgttatta tacacaaaat aatcaaagac 420 tataatcctt gatctcgatt gatcataaac aggaattgtt ttagacttca atatagtatt 480 c 481 <210> 31 <211> 459 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (459) <223> Protein related to short-chain alcohol dehydrogenases <400> 31 attagtcgta atgttggcac tatcgaagca attgctatgc gatttttatt tctgtttttc 60 aaaactccca aagaaggtgc tcaaactaat atttatctgg cagtgtctga ggaagtggaa 120 ggtgttactg gcttgtattt cactgattgt aaagtcaagg aaccatcacc gggagctcta 180 gatgacgaag cagccaagaa actgtgggat atcagcgcca agtctgtagg actggattga 240 aaaaatacca atctcattgt ggagtatgcc agtttgctgt ttgaactttc ttgtgagccc 300 aaagtggtcc ttgcatcgtg gattattgag gtggatttta tgcttcgaaa tatgaatttg 360 ttatcaagtc aaatgtgtta tctaacattc ggatgtttaa attagattcc aagctgtaaa 420 aatatgtata tttgtagctg tatgtactat ctatcataa 459 <210> 32 <211> 504 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene <222> (1) 504 <223> acyltransferase <400> 32 aagcagtggt atcaacgcag agtggccatt acggccggga agcattgcga ggataaatgg 60 tggaccaact tactgtacat caataatttt taccccgaga attttaacga ccagtgtgtt 120 ggttggacgt ggtatttggc taatgatatg caattctttg ttatatcacc aatcctgttg 180 attttggcat acaagtacgg atggcgtggg ttgatgagca gtacgggagt gttgttgttg 240 atcagttcca ttgtcattgc ggtaatcatt ggttcctatg acgtggatcc cgttgttaac 300 ctcaaagcgc ttgttcgtgg tacagatgaa cttaagaaaa agtcagaaga actcgataag 360 tatgtctaca gcaaaccgta ctgtcgcatt caaccgtatc ttgttggatt tatcctcggc 420 tatgccgtat acatgaagta tcgcgtgcct attagaagac acggttggtt gcttgctgtg 480 attggttggt ccgtggcatt tgcg 504 <210> 33 <211> 1644 <212> DNA <213> Scleronephthya gracillimum <220> <221> gene &Lt; 222 > (1) .. (1644) <223> Low-density lipoprotein receptors containing Ca2 + -binding          EGF-like domains <400> 33 aagcagtggt atcaacgcag agtggccatt acgccgggag agcacgttgg taaagaaaac 60 aaaaccaggc gttgtcagtg tttatgatta ttatgatcca gaaagtgaag cgagaataat 120 gtacaacatc acagacgaag attgttatga gcaaatggca tcatacgctg agccattcat 180 acttttctgc gcgagaaatg aaatcctggc aatggatatg gcaacttttg acacacaagt 240 tctcattcgt ggtctttata attccgtctc tgtgttggat attgatacgg tggatagaaa 300 aatgtacttt gttgatgatg aacatatata tcaggcgaat tttaacgggt cagatatgaa 360 gattgttgtg aagaatgctt atcccgagga tataacaatt gattggatag gacatcgtat 420 attttggacc aattggggtg aaaataggat tcatgtggcg aattcagatg gtaaagaaag 480 tagggtacta atcaatactc gccatccctc gtgtatagcg gtggatccaa ttgtaggatt 540 cttattttgg gctgaaactg aactctttgg ttactccacc ttgaatcgaa gacaaatgac 600 cagcaataaa ataaaaattt tgatcagatc gcgccattgg tacacaagac taattctgga 660 ccacataaac aggcgagtgt acttggccga tgtcagtggg aacgtgctgt caatggatta 720 cgaaggtgga gacagaattg caatattcgg ttatggatat ggttcgtttt atgctgtttc 780 tttcgatgtc tttggagatt caatttattt caagaaacaa ggatcatctg acattggtga 840 aatgaatgca ttcagcagaa atatttcgcg ttatatttct ccaaatattt cccatatcaa 900 ccggtttatc gtcgtcgaga aatctcggca acctgaagaa ggatgcccat cgctaccatg 960 tggtccttat ggttactgta caaatgtcaa tgattcatac tcatgtcagt gtgaattggg 1020 tttcgcatca tcagatggtg gacggtcttg tacagatatc gatgaatgca ctttgtctaa 1080 atgtaatttt acgggaattc aatcatgcca gaataaccaa ggatcttacc agtgtatatg 1140 taagaaagga tttacactta acggattgac atgttctgaa gtgtacatca gattaaatgg 1200 ttcaaataat aacagtggaa gagtggaaat ctttcatccc tcatttggct ggggtttgat 1260 atgtgatcac aaatgggata ttactgatgg tggcgtggtc tgtcgacagt taggtttcat 1320 aggagcgtta aagacgagac gtgaagacca aacaagtggt cctatgctgc ttgataaagt 1380 taaatgcacc ggtgatgaaa catacatttg ggattgcagt cacagtggat ggaatgttca 1440 tgactgtagc aatttcgaag ttgcaggcgt tgagtgttat tagcatctcg actatttgaa 1500 aataagaaga ctgaatttac aagatgacta cattttatcg aaaagtttta aatcgtattt 1560 catattatga aataatcccc cattttacag catagaatta tttttgcatt aaaattgatt 1620 tcgaaataaa aatattaatt tctt 1644

Claims (18)

As an active ingredient, at least one polynucleotide selected from the group consisting of the nucleotide sequences of SEQ ID NOS: 1 to 33 derived from the sea horse drumstick ( Scleronephthya gracillimum) ;
A complementary base sequence complementary to the polynucleotide;
A polynucleotide fragment of 20 to 200 bases consecutively located in a fragment of said polynucleotide; And
And at least one probe selected from the group consisting of a complement of the polynucleotide fragment,
Characterized in that seawater temperature is 28 to 30 占 폚, and seawater temperature rise is 7.5 to 7.8.
delete delete delete A polynucleotide selected from the group consisting of the nucleotide sequences of SEQ ID NOS: 1 to 33 derived from the Scleronephthya gracilimum as an active ingredient;
A complementary base sequence complementary to the polynucleotide;
A polynucleotide fragment of 20 to 200 bases consecutively located in a fragment of said polynucleotide; And
And at least one probe selected from the group consisting of a complement of the polynucleotide fragment,
A microarray for dual stress detection of seawater-on-rising and marine acidification, characterized in that the seawater temperature is in the range of 28 to 30 캜 and the seawater temperature is in the range of 7.5 to 7.8.
delete delete delete 6. The microarray of claim 5, wherein the microarray comprises one or more spots and at least one probe is fixed to the one spot.
A kit for detecting whether or not a sea water has risen and an ocean acidity, comprising the microarray of claim 5 or 9.
11. The detection kit according to claim 10, further comprising a mixture in which the standard nucleic acid labeled with the detectable first labeling substance and the standard nucleic acid labeled with the detectable second labeling substance are mixed in the same ratio.
delete 12. The detection kit according to claim 11, wherein the first labeling substance and the second labeling substance are the same or different from each other and are at least one selected from the group consisting of radioactive isotopes, fluorescent substances, chemiluminescent substances and enzymes. .
The method of claim 13, wherein the first labeling substance and the second labeling substance are independently selected from the group consisting of Alexa Fluor 350, Alexa Fluor 430, Alexa Fluor 488, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 594, Alexa Fluor 658, Cyclin-3, cyanine-5, fluorescein, bodipy, Texas red, FITC (Fluorescein Isothiocyanate), rhodamine, d-NTP (including d- , Reactive dyes with amino-allyl modified dNTPs, horseradish peroxidase, and biotin.
(a) A mixture of the test nucleic acid labeled with the detectable first labeling substance and the standard nucleic acid labeled with the detectable second labeling substance in the same ratio in a probe immobilized to the microarray according to the fifth or the ninth aspect Lt; / RTI &gt;
(b) measuring the hybridization degree of each of the test nucleic acid and the standard nucleic acid with respect to the probe, and comparing the hybridization degree of the test nucleic acid with the standard nucleic acid.
16. The method according to claim 15, wherein in step (a), the sample nucleic acid and the standard nucleic acid are cDNAs synthesized from the RNA of a probe of a specimen of Scleronephthya gracilimum and a RNA of a probe of a probe derived from a standard pink sea bream ( Scleronephthya gracilimum ) Detection method.
delete delete

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