CN1769454A - One group of buccal cavity white spot-related gene - Google Patents

Abstract

The present invention about one group in the buccal cavity white spot process, express to continue the enhanced gene.In the buccal cavity white spot process, expressing lasting enhanced gene has 13, and it has the nucleotide sequence shown in SEQID:1, SEQID:2, SEQID:3, SEQID:4, SEQID:5, SEQID:6, SEQID:7, SEQID:8, SEQID:9, SEQID:10, SEQID:11, SEQID:12, SEQID:13.The disclosed genes involved of the present invention, its advantage shows: carry out the dynamic follow-up study by the gene expression profile to the buccal cavity white spot whole process, filter out the relevant target gene group of leukoplakia cancer process, can hold leukoplakia cancer mechanism fast, comprehensively from integral level.

Description

One group of buccal cavity white spot-related gene

Technical field

The present invention expresses lasting enhanced gene about the covert correlation gene of oral carcinoma especially in regard to buccal cavity white spot.

Background technology

Oral squamous cell carcinomas accounts for 4% of whole body malignant tumour, because the oral cavity is positioned at respiratory tract and gastral critical positions, pathology result has a strong impact on patient's life and life quality.The generation development mechanism of research oral squamous cell carcinomas is the key of fundamentally preventing curing oncoma.

Oral leukoplakia is one of main source of oral squamous cell carcinomas, and for the leukoplakia cancer Mechanism Study, Chinese scholars is used traditional pathology, immunohistochemical methods, Electronic Speculum and Protocols in Molecular Biology more single or multiple genes are studied.Along with the appearance of biochip technology, at present, existing many scholar's application chip technology are carried out gene expression profile research to bladder cancer, mammary cancer, oral carcinoma, esophagus cancer etc., and new discovery is had the target spot of using potential carry out patent protection.

Traditional molecular Biological Detection means are once tested and are only detected one or several gene, can not totally hold the relevant target gene group's of leukoplakia cancer effect.Though existing scholar uses chip technology that oral carcinoma, esophagus cancer etc. is carried out gene expression profile research, because oral leukoplakia canceration process is longer, and its canceration uncertainty, be difficult in clinical patients, do the dynamic follow-up study.Therefore, the research shortage system for the suspicious gene of buccal cavity white spot process reports.And chip data treatment process Heirarchical clustering at present commonly used is big in the chip data amount, under the data complexcase, regular meeting causes inaccurate even wrong cluster to influence interpretation of result.

Summary of the invention

The objective of the invention is to: provide one group in the buccal cavity white spot process, express lasting enhanced gene.

The objective of the invention is to realize: use biochip technology the gene expression profile of leukoplakia cancer whole process to be carried out the dynamic follow-up study, thereby hold the relevant target gene group of leukoplakia cancer on the integral level in conjunction with the CPP-SOM data analysing method by following technological method.The present invention has the foundation and the sample collection of following steps (1) 4NQO inductive rat leukoplakia cancer model; (2) the total RNA of sample extracts and chip hybridization; (3) chip scanning and data analysis.

In the buccal cavity white spot process, expressing lasting enhanced gene has 13, and it has the nucleotide sequence shown in SEQID:1, SEQID:2, SEQID:3, SEQID:4, SEQI D:5, SEQID:6, SEQID:7, SEQID:8, SEQID:9, SEQID:10, SEQID:11, SEQID:12, SEQID:13.The albumen of SEQID:1 genes encoding is chemokine (C-X-C motif) aglucon 9, the albumen of SEQID:2 genes encoding is cyclin A2, the albumen of SEQID:3 genes encoding is to be similar to cyclin B2, the albumen of SEQID:4 genes encoding is estrogen receptor 2, the albumen of SEQID:5 genes encoding is Janus kinases 3, the albumen of SEQID:6 genes encoding is chemokine C-X-C aglucon 10, the albumen of SEQID:7 genes encoding is vehicle 1, ATP-binding cassette, the B subfamily, the albumen of SEQID:8 genes encoding is cell division cycle 2 homologue A, the albumen of SEQID:9 genes encoding is nuclear factor KB p105 subunit, the albumen of SEQID:10 genes encoding is membrane-spanning domain 4, the A subfamily, 7.

The disclosed one group of buccal cavity white spot-related gene of the present invention, its advantage shows:

1, carries out the dynamic follow-up study by gene expression profile, filter out the relevant target gene group of leukoplakia cancer process, can hold leukoplakia cancer mechanism fast, comprehensively from integral level to the buccal cavity white spot whole process;

2, can promote the raising of levels such as oral leukoplakia diagnosis and treatment and prognosis judgement;

3, based on this, design and develop purposes biochips such as oral precancerous lesion diagnosis, judging prognosis, further drug target is excavated in research, for individuation pharmacological agent provides foundation.

Description of drawings

Fig. 1 shows that total RNA extracts the result

Fig. 2 shows the changing conditions of CPP-SOM Dynamic Display mile abnormality proliferative phase gene, and on behalf of gene, black raise, and white is represented the gene downward modulation

Fig. 3 shows the changing conditions of CPP-SOM Dynamic Display moderate paraplasm stage gene, and on behalf of gene, black raise, and white is represented the gene downward modulation

Fig. 4 shows the changing conditions of CPP-SOM Dynamic Display severe paraplasm stage gene, and on behalf of gene, black raise, and white is represented the gene downward modulation

Fig. 5 shows the changing conditions of CPP-SOM Dynamic Display carcinoma in situ stage gene, and on behalf of gene, black raise, and white is represented the gene downward modulation

Fig. 6 shows the changing conditions of CPP-SOM Dynamic Display squama cancer stage gene, and on behalf of gene, black raise, and white is represented the gene downward modulation

Embodiment

Below in conjunction with embodiment the present invention is further described.

Embodiment 1

1.14NQO the foundation and the sample collection of inductive rat leukoplakia cancer model

1.1-1 animal and material: 20 of Wistar male rats, body weight 220 ± 10g, 160 days ages of mouse, per 4 one cages, 23 ± 2 ℃ of room temperatures, humidity 30-50%, the refining particles feed, dark cycling condition of 24 little time is raised down.

4NQO: U.S. Sigma company produces, and makes 0.0002% concentration solution with distilled water and keeps in Dark Place standby for 4 ℃.

1.1-2 grouping: 35 rats are divided into 2 groups by table of random number, 5 of normal control groups, and ordinary tap water is fed; 17 of experimental group, the 4NQO nursing of drinking water; Respectively at putting to death in 9 weeks, 13 weeks, 16 weeks, 20 weeks, 24 weeks, 32 weeks.Observe variation substantially, leave and take the tongue sample.1/3 sick damage place behind the back is cut open from center line, and half puts in the 10% neutral formalin solution fixing, routine paraffin wax embedding, 4um section, HE dyeing back om observation.Half places RNAlater (Qiagen company) ,-80 ℃ of preservations

1.1-3 histopathological examination

Case definition: according to 12 Case definition of WHO oral precancerous lesion cooperation center (1978) sample is carried out grading diagnosis (comprising normal mucous membrane, mile abnormality hyperplasia, moderate paraplasm, severe paraplasm, carcinoma in situ, squama cancer), all diagnosis all adopt double-blind method to confirm.

Collect healthy tissues 5 examples, mile abnormality hyperplasia 4 examples, moderate paraplasm 6 examples, severe paraplasm 6 examples, carcinoma in situ (24 week) 4 examples, squama cancer (32 week) 5 examples.

1.2 the total RNA of sample extracts and chip hybridization

1.2-1 under RNase free (the removing the RNA enzyme) condition, (step 1. cryopreserved tissue of Trizol reagent single stage method adds 1ml trizol, homogenizer homogenate, room temperature placement 5 minutes to use the Trizol method; 2. add the 0.2ml chloroform, shook 15 seconds; 3.4 centrifugal 15 minutes of degree 12000rpm; 4. get supernatant, add the 0.5ml Virahol, shake up room temperature and placed 10 minutes; 5.4 centrifugal 10 minutes of degree 12000rpm; 6. abandon supernatant, add 1ml 75% washing with alcohol RNA in precipitation, centrifugal 5 minutes 7. seasoning RNA of 4 degree 7500rpm are dissolved in packing in methane amide or the pure water.) the total RNA of extraction sample.Sample detects the absorbance A value through spectrophotometer.Detect 28S, the total RNA of 18S band variation .Rneasy test kit (Qiagen company) purifying through agarose gel electrophoresis.

1.2-2 the preparation of probe and chip hybridization: chip used is Amersham commercialization chip-codelink Uniset Rat I Bioarry.

Utilize the CodeLink test kit to prepare probe, the external cRNA mark vitamin H that is inverted to after the synthetic cDNA two strands.With T7 primer and SuperScript II reversed transcriptive enzyme and the total RNA of 10 μ g at 1 hour synthetic first chain cDNA of 42 ℃ of hatchings.The first chain cDNA mixes with E.coliDNA polysaccharase I and RNaseH, and at 2 hours synthetic second chain cDNA of 16 ℃ of hatchings, the double-stranded cDNA of formation uses QIAquick PCR purification kit (Qiagen company) purifying.CRNA with t7 rna polymerase and biotin-11-UTP in-vitro transcription synthesizing biotinylated mark.With RNeasy test kit (Qiagen company) purifying cRNA.Use 20 minutes fragmentation cRNA of 94 ℃ of heating of 5 * fragmentation buffer (segment damping fluid).CRNA behind the fragmentation and Uniset bioarry are positioned on the shaker that CodeLink provides, and shaking table is adjusted rotating speed 300rpm, and 37 ℃ of hatchings are spent the night.0.75 46 ℃ of washing hatchings of * TNT buffer 1 hour.Then, lucifuge was hatched 30 minutes under use Cy5-Streptavidin (Cy5-streptavidin) room temperature.1 * TNT washs chip 4 times (each 5 minutes), and the flushing chip is after 30 seconds in 0.1 * SSC/0.05%TweenTM, and the centrifugal 3min of 2000rpm dries.

1.3 chip scanning and data analysis

With Array Scanner (Axon company) scanning chip.Carry out picture processing and standardization of data with codelink expressionanalysis software.Adopt the CPP-SOM (self organization map binding constituents plane methods of exhibiting) that is based upon on the Matlab 5 software package bases to carry out the further analysis and the displaying of data then.

2. result

2.1 total RNA extracts the result

Total RNA ultraviolet spectrometry degree OD260/OD280:Ratio value is between 1.7-2.0.

Total RNA electrophoresis: 28S, 18S band clearly, and the brightness of 28S band is the twice of 18S band.(seeing that Fig. 1 shows).

On behalf of gene, the changing conditions of CPP-SOM Dynamic Display different steps gene, black raise, and white is represented the gene downward modulation.(seeing that Fig. 2,3,4,5,6 shows).

2.3 difference expression gene analysis

In the buccal cavity white spot process, expressing lasting enhanced gene has 13 (table 1).

Table 1 part is expressed the gene that significantly increases

Numbering	The gene title	SEQ	ACCN	Uigene
					1	Chemokine (C-X-C motif) ligand 9 (Cxc19) (chemokine (C-X-C motif) aglucon 9)	SEQID：1	NM_145 672	Rn.7391
2	Cyclin A2 (cyclin A2)	SEQID：2	NM_053	Rn.1309

			702.	4
					3	Similar to cyclin B2 (being similar to cyclin B2)	SEQID：3	BE1116 88	Rn.6743
4	Estrogen receptor 2 (estrogen receptor 2)	SEQID：4	NM_012 754.1	Rn.3746 0
					5	Janus kinase 3 (Janus kinases 3)	SEQID：5	NM_012 855.1	Rn.1115 9
6	Chemokine (C-X-C motif) ligand 10 (chemokine C-X-C aglucon 10)	SEQID：6	NM_139 089.	Rn.1058 4
					7	Transporter 1, ATP-binding cassette, sub-family B (MDR/TAP) (vehicle 1, ATP-binding cassette, the B subfamily)	SEQID：7	NM_032 055.	Rn.1076 3
8	Cell division cycle 2 homolog A (S.pombe) (cell division cycle 2 homologue A)	SEQID：8	NM_019 296.1	Rn.6934
					9	Nuclear factor kappa B p105 subunit (nuclear factor KB p105 subunit)	SEQID：9	L26267	Rn.2411

10	Membrane-spanning 4-domains, subfamily A, member 7 (predicted) (membrane-spanning domain 4, the A subfamily, 7)	SEQID：10	XM_215 200.	Rn.4403 7
					11	The unknown function gene	SEQID：11	AA9451 00	Rn.1057 75
12	The unknown function gene	SEQID：12	AA9579 92	Rn.9464
					13	The unknown function gene	SEQID：13	AI1784 76	Rn.1654 6

SEQUENCE LISTING

＜110〉No.9 People's Hospital Attached to Shanghai No.2 Medical Univ

＜120〉one group of buccal cavity white spot-related gene

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＜213〉Rattus norvegicus (norway rat)

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gcctccggag tcttctgagg cttgcatcac caagctccag gcttggtgct gccgtctggc 780

tgctggtgag ctagactttc agcagctgca ttttgagtca cccttgagcc cctgcacaaa 840

gtgtgtctca gagatccgta caaatggtct gggggttcca ccagggttgg agaccaccag 900

ggttggcttc cagaaccttc catgtagctt tttgagacca tggaatttca tcattaattt 960

gatcccatgt tcaaggctta ttccacattt gcctcttaaa taaaaatccc ctagacggtt 1020

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cacgagccct gcggacccgt gtatactgcc cttcagcact agtggagcgg tcttcctaaa 120

aattctaggg tcataccaag cagtgccccg cgtgtcgact tcggccatgt acctttccct 180

ccacttttaa agccaggcat tgcagtagga tatagatccc ttccccctca ccatctgctt 240

cacagattac tacttaaggg aaggacataa agcaagtatt ctgcttggta aattccgcca 300

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ccctaacttt agtctttgat tagtttggaa ttccaatccg ttgcatttat agcctttgta 420

gaaatggctg tgacaaatag gtttactcac cttacctgga ctttgactca tgaacttaca 480

tttttcttaa gagtatttgg aaataaaagt tggtacccac agggcaagaa gcaaagtatt 540

aaaaaagaat tccttctctt taggttgcgc ctcttaagga ccttcctata aacgatgagc 600

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ccccatgcta tggatatatc tattgtctta gaagacgaaa agccagtgaa tgttaatgaa 960

gtacctgact atcatgaaga tattcacacg taccttaggg aaatggaggt taaatgtaaa 1020

cctaaagtga gttatatgaa gaggcagcca gacatcacta acagtatgcg ggccatcctt 1080

gtggactggc tagttgaagt gggagaagaa tataaactac agaatgagac cctgcatttg 1140

gctgtgaact acattg tag attcctctcc tccatgtctg tgttaagagg aaagcttcag 1200

cttgtgggca cagctgctat gctgctagct tcaaagtttg aagaaatata ccccccagaa 1260

gtagcagagt ttgtgtatat tacggacgat acctattcca agaagcaggt tctgagaatg 1320

gagcaccttg tattgaaagt ccttgctttt gacttggctg caccaacagt aaatcagttc 1380

cttacccagt acttcctgca cctgcagcct gccaactgca aggtagaaag cttagcgatg 1440

tttttgggag agttaagttt gatagatgct gacccatacc tcaagtattt gccatcgctt 1500

attgctggag ctgccttcca cttagctctc tacacagtca caggacaaag ctggcctgag 1560

tcattggtac aaaagactgg atatacccta gaaagtctta agccttgtct catggacctt 1620

caccagacct acctcaaagc accacagcat gcccaacagt caataaggga aaagtacaag 1680

cattcaaaat atcacagtgt ttctcttctc aacccaccag agacactaaa tgtgtgagag 1740

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taatttttaa ttggtttgtt aacattggtg ctgccaactg tcaagataat aagaataaat 1980

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ttggtgctgc caactgtcag gataataaga ataaattgat tcagaaagca aaaaaaaaaa 2100

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tttttttttt tttttttaag aaaatcaata gctttatttt tcatcaatac acaattagaa 60

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aagcccggct gcctagggtc tgcccatcag aggggaggcc aggtctttga tggttctgga 180

gttcagctga gggatcgtgc tgatcttcag gagcctgctg ctggcatatt tgttcttgac 240

agcgatgaac ttggtgaggt tttcattgac tttcacgacg ttcttggcca tgtgctgcat 300

gacttccagt atttcactct ccatgtagcc tgtgtaatac tgctgcttta aattccattt 360

tccttggccc agcaccttct gagagaggca ggaagcagca gctgcgacct gagaagggtg 420

gtaatgcacc atgtcgtaat cgacgagggt cagctccatc aggtacttgg ctaacgtgtg 480

ctgttcaaca tccacctccc cggcttttga ggctcgcctt aagaagtgga gtggcaaagg 540

ccggcccaac tcgaatttca actctttcaa aatcagagtc tccatttctc ggatttggga 600

actggtgtaa gca 613

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＜213〉Rattus norvegicus (norway rat)

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cgggggagct ggcccagggg gagcggctgg tgctgccact ggcatcccta ggcacccagg 60

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gctctgtggc tgaggaaagc acctgtctgc atttagagaa tgcaaaatag agaatgttta 180

cctgccagtc attacatctg agtcccatga gtctctgaga acataatgtc catctgtacc 240

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agcttagtgg gagcagttgt gccagccctg ttactagtcc aaacgcaaag agggatgctc 720

acttctgccc cgtctgcagc gattatgcat ctgggtatca ttacggcgtt tggtcatgtg 780

aaggatgtaa ggcctttttt aaaagaagca ttcaaggaca taatgattat atctgtccag 840

ccacgaatca gtgtaccata gacaagaacc ggcgtaaaag ctgccaggcc tgccgacttc 900

gcaagtgtta tgaagtagga atggtcaagt gtggatccag gagagaacgg tgtgggtacc 960

gtatagtgcg gaggcagaga agttctagcg agcaggtaca ctgcctgagc aaagccaaga 1020

gaaacggtgg gcatgcaccc cgggtgaagg agctactgct gagcaccttg agtccagagc 1080

aactggtgct caccctcctg gaagctgaac cacccaatgt gctggtgagc cgtcccagca 1140

tgcccttcac cgaggcctcc atgatgatgt ccctcactaa gctggcggac aaggaactgg 1200

tgcacatgat tggctgggcc aagaaaatcc ctggctttgt ggagctcagc ctgttggacc 1260

aagtccggct cttagaaagc tgctggatgg aggtgctaat ggtgggactg atgtggcgct 1320

ccatcgacca ccccggcaag ctcattttcg ctcccgacct cgttctggac agggatgagg 1380

ggaagtgcgt agaagggatt ctggaaatct ttgacatgct cctggcgacg acgtcaaggt 1440

tccgtgagtt aaaactccag cacaaggagt atctctgtgt gaaggccatg atcctcctca 1500

actccagtat gtaccccttg gcttctgcaa accaggaggc agaaagtagc cggaagctga 1560

cacacctact gaacgcggtg acagatgccc tggtctgggt gattgcgaag agtggtatct 1620

cctcccagca gcagtcagtc cgactggcca acctcctgat gcttctttct cacgtcaggc 1680

acatcagtaa caagggcatg gaacatctgc tcagcatgaa gtgcaaaaat gtggtcccgg 1740

tgtatgacct gctgctggag atgctgaatg ctcacacgct tcgagggtac aagtcctcaa 1800

tctcggggtc tgagtgcagc tcaacagagg acagtaagaa caaagagagc tcccagaacc 1860

tacagtctca gtgatggcca ggcctgaggc ggacagacta cagagatggt caaaagtgga 1920

acatgtaccc tagcatctgg gggttcctct tagggctgcc ttggttacgc accccttacc 1980

cacactgcac ttcccaggag tcagggtggt tgtgtggcgg tgttcctcat accaggatgt 2040

accaccgaat gccaagttct aacttgtata gccttgaagg ctctcggtgt acttactttc 2100

tgtctccttg cccacttgga aacatctgaa aggttctgga actaaaggtc aaagtctgat 2160

ttggaaggat tgtccttagt caggaaaagg aatatggcat gtgacacagc tataagaaat 2220

ggactgtagg actgtgtggc cataaaatca acctttggat ggcgtcttct agaccacttg 2280

attgtaggat tgaaaaccac attgacaatc agctcatttc gcattcctgc ctcacgggtc 2340

tgtgaggact cattaatgtc atgggttatt ctatcaaaga ccagaaagat agtgcaagct 2400

tagatgtacc ttgttcctcc tcccagaccc ttgggttaca tccttagagc ctgcttattt 2460

ggtctgtctg aatgtggtca ttgtcatggg ttaagattta aatctctttg taatattggc 2520

ttccttgaag ctatgtcatc tttctctctc tcccg 2555

<210>5

<211>3778

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＜213〉Rattus norvegicus (norway rat)

<400>5

gccttgaact tctaatcttc cgcctccatc tctggagttt tctggtaaca agctatactc 60

atggcacctc caagcgagga gacgcctctc atctctcagc gctcttgtag cctctcatcc 120

tcagaagcag gagccctgca tgtgctcctt cctccccggg gacctgggcc tccccagcga 180

ctgtcattct cttttgggga ctacttggct gaggatctgt gtgtgcgggc tgccaaggcc 240

tgtggcatcc tgcccgttta tcattcgctc ttcgctctgg ccaccgagga cttgtcttgc 300

tggttccccc caagccacat cttctccata gaggacgtgg acactcaagt cttggtctac 360

aggctacgct tttatttccc tggttggttt ggactggaga cctgtcaccg ctttgggtta 420

cacaaagatt tgaccagtgc catccttgac gtacatgttt tagaacacct ctttgcccag 480

caccgcagtg acctggtgag tgggcgtctc ccagtaggcc ttagcctgaa ggaccaggga 540

gagttcctga gcctagctgt gctggacctg gcccagatgg ctcgtaagca ggcccagcgg 600

ccaggagagc tgctgaagtc tgtcagttac aaagcctgtc tgccgcccag cctgcgcgac 660

ctgatccagg gccagagctt cgtgacgcgc aggcgcatcc gcaggaccgt ggtccaggcg 720

ctggcgccgt gtagtagcct gccaagccga ccctacgcac tcatggccaa gtacatccta 780

gaccttgagc ggctgcatcc agcggccacc actgagtcct tcctcgtggg actcccgggc 840

gcccaggagg agccgggttg cctgcgtgtg acaggggata atggcatcgc ctggagctcc 900

aaggaccagg agcttttcca gaccttctgt gactttcccg aaatcgtgga tgtcagcatc 960

aagcaggctc cacgcg ggg tccggcaggg gagcaccggc tagtcaccat caccaggatg 1020

gacggtcaca tcctggaagc ggagttcccg gggctgcctg aggcgctgtc tttcgtggcg 1080

ctcgtggatg ggtactttcg cctgatctgc gactccaggc atttcttctg caaggaggtg 1140

gcaccgccac ggctgctgga ggaggaggcg gagctgtgcc atggacccat cacgttagac 1200

tttgccatcc acaagctgaa ggctgctggc tccctcccgg gctcctacat tctccgacgc 1260

agcccgcagg actatgacag ctttcttctt actgcctgtg tccagactcc tcttggcccc 1320

gactacaagg gctgcctcat ccgccaagac cccagtgggg ctttctccct ggttggtctc 1380

agccagctgc acagaagcct acaggagctg ctcacagcct gctggcattc tggactgcaa 1440

gtagacggca ctgccctgaa cctcacatct tgctgcgtcc ccagacccaa ggaaaagtcc 1500

aatttgatcg tggtgcgtag gggccgcaac cccacgcctg cccccgggca ctccccatcc 1560

tgctgtgcgc tgactaagtt gagcttccac acaatcccag cagacagcct ggagtggcac 1620

gagaatcttg gtcacggctc ttttaccaag atcttccatg gccataggcg ggaggttgtg 1680

gatggtgaga cacatgacac agaagtcctc ctgaaggtta tggactccag acaccagaac 1740

tgcatggagt cctttctgga agccgcaagc ttgatgagcc aagtatccta cccgcacctg 1800

gtgttgctgc atggagtctg catggctgga gacagcatca tggtgcagga atttgtgtac 1860

cttggagcaa ttgacacgta cctgcgcaag cgtggtcacc tggtgccggc cagctggaaa 1920

ctgcaggtga ccaagcagct ggcctatgcc cttaactact tggaggacaa aggcctccct 1980

catggcaacg tctcagcgag gaaggtgctg ctagctcgtg agggggttga tgggaatcca 2040

cctttcatca agctgagtga tcctggtgtc agcccaactg ttttgagcct ggaaatgctc 2100

actgacagga ttccctgggt ggccccggag tgtctccagg aagctggaac gctcaacttg 2160

gaggctgaca agtggggctt tggcgccacc acgtgggagg tgttcagcgg ggcacccatg 2220

cacattacct cgctggagcc cgccaagaag ctgaagttct atgaggaccg gggacagctg 2280

ccggccctca aatggacaga gctggagggg ctcatcgcac agtgcatggc gtatgatcct 2340

ggccggcgcc cctccttccg agccatcctc agagacctca acggcctcat tacatcagat 2400

tacgagctcc tctcagaccc cacacctggc atcccgaatc cccgagatga gctgtgtggt 2460

ggtgcccagc tctatgcctg ccaggacccc gccatcttcg aggagagaca ccttaagtac 2520

atctccctgc tgggcaaggg gaacttcggc agcgtggagc tgtgccgcta cgaccctctg 2580

ggagacaata cgggacccct ggtagcagtg aagcagttac agcacagcgg gccagaacag 2640

cagagggact tccagcgaga gattcagatc ctcaaggctc tacactgtga cttcatcgtc 2700

aagtaccggg gagtcagcta tgggcctggt cgccaggagc tgcggttagt gatggagtac 2760

ctgcccagcg gctgcctgcg ggacttcctg cagcgccacc gcgcgcgcct gcacaacgac 2820

cgcctactgc tgttcgcctg gcagatctgc aagggcatgg agtacctggg cgcacgccgc 2880

tgcgtgcatc gtgacctggc tgcgcgcaac atcctggtgg agagtgaggc gcatgtgaag 2940

attgctgact tcggcctcgc taagctgctg cctcttggaa aagactacta cgtggtccgc 3000

gtgccgggcc aaagtcccat cttctggtac gcccctgagt ccctatctga caacatcttc 3060

tcccgccaat ctgacgtgtg gagcttcggc gtggtgctgt atgagctctt cacctacagc 3120

gacaagagct gcagcccatc cactgagttc ctgcgtatga ttgggcctga gcgtgaggga 3180

tccccgctct gccacctctt ggagctgctg gcagagggcc gccgtctccc accaccctcc 3240

acctgcccta ctgaggttca agagctcatg cagctgtgct ggtcgcccaa cccacaagac 3300

cggccagcgt ttgacaccct gagcccccag ctggatgcgc tgtggcgcgg aagccccgga 3360

tagcaagcag ccagggatga gcgtgagttt ggttcctatg atctggctgt gtgacttcca 3420

ggcgggaagc tgtccctctc tgggccccag cacccccaat cccctctggc cacctcttcc 3480

catcattctt ccctcccaga atggggatgt taaataagtg accccaggct ctgcacggtt 3540

gtcattacct ctggaaatgc tttactagca agtctgcctt ctgcagaact cacaacccca 3600

ggtcccaaga gcaccctaga cttggccaga aacccaccca gacccccaga ccctctcttg 3660

accctaggtc tgtgcccagt cccttgctct cctgtccatg ggtttcttgg gggctgtgaa 3720

acctcagcat ggtctcctga aagattgggt tctgggagtg tttgttgaat gaataaat 3778

<210>6

<211>1133

<212>DNA

＜213〉Rattus norvegicus (norway rat)

<400>6

cctcggctga gctgcattcc aatcccagct acatccggag cccagctaca ttcgagccca 60

gccacatccc gagccaacct tccagaagca ccatgaaccc aagtgctgct gtcgttctct 120

gcctcgtgct gctgagtctg agtgggactc aagggatccc tctcgcaaga acggtgcgct 180

gcacctgcat cgacttccat gaacagacgc tgagacccag ggccatagga aaacttgaaa 240

tcattcctgc aagtctatcc tgtccgcatg ttgagatcat tgccacaatg aagaagaaca 300

atgagaagag gtgtctgaat ccggaatctg aggccatcaa gagcttattg aaagcggtga 360

gccaaagaag gtcaaaaaga gctccgtaac tagagagaag ccactcgcca cagtgctgag 420

accgatggac agcagagaga cggtctctcc acctcccttt acccagtgtg cggctagtcc 480

taactgtccc tgtttctcct gaccatggtc ccatcagctg gtactcccac tacagcgtga 540

tggacaaggc ctggtcctga gacaaaagta actccagcag caaggcttcc caattctcta 600

agagctggtc cgaatcttcc ctcaggcagc tatgacggct ctcctagctc tgttccgtaa 660

gctatgtgca ggtactaatc tcttcagcat gtgccatgcc ccagcctgct ccacacaccc 720

tccttctccc tagctctaag ctcatcagtt ctgagttcac ctgagctcct ttatttcaaa 780

tgcagtccag gtgagatggc aaatcaagtt tgtcagaaca aacttaccac caccttccca 840

agggaatttc ataactcaga atactcacag gaacctagac atgcatgttt aaatattatt 900

taatgaccga ctgtacaaag tggaactcct agatgtattt tttgtacgat tttcattgta 960

tatgtaagaa cttgtgtggt taagtatgta tcaatgggta gttaaagttt acataggcaa 1020

atgctttgaa tgctacatat tacaagatgt gttggatggt tttcaaaata aaatgtactg 1080

tattgaatgt agtatgagac aaaaaagtaa taaagtaata ataactgaca tga 1133

<210>7

<211>2178

<212>DNA

＜213〉Rattus norvegicus (norway rat)

<400>7

atggctgcgc acgcctggcc gacggccgcc ttgctgctgc tgctggtgga ctggctgctg 60

ctgcggtccg tgctcccggg aatcttctcc ctgttggttc ccgaggtgcc gctgctccgg 120

gtctgggccg tgggcctgag tcgctgggct atcctgggac taggggtccg cggggtcctc 180

ggggtcaccg cgggagcccg tggctggctg gctgctttgc agccactggt ggcggcgctg 240

ggtttggccc tgcctggact tgcctcgttc cgaaagctgg ccgcctgggg agcactccgg 300

gagggtgaca acgctggact gctccactgg aacagtcgct tagatgcctt cgttctcagt 360

tatgtggccg cattgcccgc agctgccctg tggcacaagt tggggagctt ctgggcgccc 420

agcggccaca agggcgctgg agacatgctg tgtcggatgc taggcttcct ggactccaag 480

aaggggcgtc tccacctggt tctggttctc ttgatcctct cctgccttgg ggaaatggcc 540

attcccttct tcacaggccg catcactgac tggatccttc aggataagac agcccccagc 600

ttcgcccgca acatgtggct catgtgtatt cttaccatag ccagtacagc gctggagttt 660

gcaggagatg gaatctacaa catcaccatg ggccacatgc acagccgcgt gcatggagag 720

gtgtttcggg ccgtccctaa ccaggagaca ggatttttcc tgaagaaccc aacaggttcc 780

atcacatctc gggtgactga ggacacctcc aacgtgtgcg agtccattag tgacaagctg 840

aacctgttcc tgtggtacct ggggcgaggc ctgtgtctcc tggcgttcat gatttggggg 900

tcattctacc tcactgtggt caccctgctc agcctgcctc tgcttttcct tctgcccagg 960

aggctgggga aagtgtacca gtcactggca gtgaaggtgc aggagtctct agcaaagtcc 1020

acgcaggtgg ccctcgaggc cctgtcggcg atgcctaccg tacggagctt tgccaacgag 1080

gagggagagg cccagaagtt taggcagaag ttggaagaaa tgaagccgct aaacaagaaa 1140

gaggccttgg cttacgtcac tgaagtctgg accatgagtg tctcgggaat gctgctgaag 1200

gtgggaattc tgtacctcgg tgggcagctg gtggtcagag gggctgtcag cagtggcaac 1260

ctcgtctcct ttgttctcta ccagcttcag ttcaccaggg ccgtggaggt cctgctctcc 1320

atctatccct ccatgcagaa gtccgtgggc gcctcggaga aaatattcga atacctggac 1380

cggactccct gctctccgct cagtggctca ctggcacctt taaacatgaa aggcctcgtc 1440

aagttccaag atgtctcctt tgcctaccca aaccatccca acgtccaggt gcttcagggg 1500

ctgactttta cgctgtatcc cgggaaggtg acctccttgg tgggacccaa tgggtcaggg 1560

aagagcaccg tggccgccct gctgcagaac ctgtaccagc ccaccggggg caaggtgctc 1620

ctggatggcg agcccctggt ccagtatgat caccactacc tgcacacgca ggtggccgca 1680

gtgggacaag agccactgct atttggaaga agttttcggg aaaatattgc ctatggcctg 1740

actcggactc caaccatgga ggaaatcaca gctgtggcca tggagtctgg agcccacgat 1800

ttcatctctg gattccctca gggctatgac acagaggtag gtgaaactgg gaaccagctg 1860

tcaggaggtc agcgacaggc ggtggccttg gctcgagcct tgatccggaa gccacgcctg 1920

cttatcttgg acgatgccac cagtgccctg gatgctggca accagctacg ggtccagcgg 1980

ctcctgtatg agagcccgga gtgggcctct cggacggttc ttctgatcac ccagcagctc 2040

agcctggcag agcgggccca ccacatcctc ttcctcaaag aaggctctgt ctgcgagcag 2100

ggcacccacc tgcagctcat ggagagagga gggtgttacc ggtccatggt ggaggctctt 2160

gcggctcctt cagactga 2178

<210>8

<211>1184

<212>DNA

＜213〉Rattus norvegicus (norway rat)

<400>8

ctcggcctct aagctcccgg agtcgctgcg tccgcgcagt ccggaactcg tgtggccacc 60

gcgggaacag agagggtccg ttgtaacctg ttgagtaact atggaggact atatcaaaat 120

agagaaaatc ggagaaggga cttatggtgt ggtatataaa ggtagacaca ggactactgg 180

ccagatcgtg gccatgaaga aaatcagact tgaaagcgag gaagaaggag tgcccagtac 240

ggcaatccgg gaaatctctc tgttaaaaga acttcgacat ccaaatatag tcagcctgca 300

ggatgtactc atgcaggact ccaggctgta tctcatcttc gagttcctgt ccatggatct 360

caagaagtac ctggactcta tccctcctgg ccagttcatg gattcttcgc tcgttaagag 420

ttacttgtac caaatcctcc aggggattgt gttttgtcac tcccgacgag ttcttcacag 480

agacttgaaa cctcaaaatc tattgattga tgacaaagga acaatcaaac tggcagattt 540

cggccttgcc agagcgtttg gaataccgat aagagtgtac acacacgagg tagtgacgct 600

gtggtaccgg tcgccagagg tgttgctggg ctcggctcgc tactccaccc cggttgacat 660

ctggagcata gggaccatat ttgcagagct ggcgaccaag aagccgcttt tccacggcga 720

ctcagagatt gaccagatct tcaggatctt cagagctctg ggcaccccta acaacgaagt 780

gtggccagaa gtagagtccc tgcaggacta caagaacact ttccccaagt ggaagccggg 840

gagcctcgcg tcccacgtca agaacctgga tgaaaacggc ttggacttgc tctcgaaaat 900

gctggtctat gatccagcca aacgaatctc tggaaaaatg gcccttaagc acccatactt 960

tgacgacttg gacaatcaga tcaagaagat gtaaccctct ggaagtcccg ccctgctgtg 1020

gcagggaaga tcgtggcgtt tactgttggc tcttcctgtc ttgtatagtt ttcttttctt 1080

tgtctgtaaa ccatcatctg gacttttcct attttcctac gtataactta attaacatgt 1140

aaatattatt ccatatgaat ttaaatataa ttctgtatat gtgt 1184

<210>9

<211>2245

<212>DNA

＜213〉Rattus norvegicus (norway rat)

<400>9

agggagattc tgaatccccc tgagaaagaa acacaaggtg aggggcccag cctgttcatg 60

gccagcacca agaccgaagc aattgccccg gcatccacca tggaagacaa ggaagaggat 120

gtggggtttc aggataacct ctttcttgag aaggcgctgc agctcgccaa gcggcatgcc 180

aacgccctct tcgactacgc ggttacggga gatgtgaaga tgctgctggc ggtacagcgc 240

catctcaccg cggtgcagga tgagaacggg gacagtgtcc tacacttagc catcatccac 300

cttcatgctc agctcgtgag ggatctgcta gaagtgacat ctggttcgat ctctgatgac 360

atcatcaaca tgagaaacga tctgtatcag acacctttgc acttggccgt gatcaccaag 420

caggaagatg tggtggagga cttgctgagg gttggggctg acctgagtct tctggaccgc 480

tggggcaact ctgtcctgca cctagctgcc aaagaaggac atgataaaat cctcggggtc 540

ctactcaaga acagcaaggc agcactcctt atcaaccacc ccaacgggga aggtctgaat 600

gccattcaca tagctgtgat gagcaacagc ctgtcatgtc tgcagctgct tgtggctgct 660

ggggcagaag tcaacgctca ggagcagaag tctgggcgca cggcactgca cctggccgtg 720

gagtacgaca acatctcgtt ggccggctgc ctgctcctgg agggtgacgc cctcgtggac 780

agcaccacct acgatgggac gacacctcta cacatagcgg ctggaagagg gtccaccagg 840

ctggcagctc ttctcaaagc agcaggagca gaccccctgg tggagaactt tgagcctctc 900

tatgacctgg acgactcttg ggagaaggct ggagaagatg agggagtagt gccaggcacc 960

acacccctgg acatggctgc caactggcag gtatttgaca tactaaatgg gaaaccatat 1020

gagccagtgt tcacatctga tgacatactc ccacaagggg acattaagca gctgacagaa 1080

gacacgaggc tgcaactctg caagctcttg gaaattcctg atcccgacaa gaactgggcc 1140

actctggcgc agaagttagg tctggggata ttgaacaatg ccttccggct gagtcccgcc 1200

ccttctaaaa ctcttatgga caactatgag gtctctgggg ggaccatcaa agagctggtg 1260

gaggccctgc gacagatggg ctacacagag gccattgaag tgatccaggc agccttccgc 1320

accccggaaa ccacagcctc cagccccgtg accactgctc aggcccactt gctgcctctc 1380

tcgtcctcct ccacaagaca gcacatagat gagctccgag ataatgacag cgtgtgtgac 1440

agtggtgtgg agacatcctt ccgcaaactc agcttctcgg agtccctcac tggagatggc 1500

ccactgctat ctctgaacaa aatgcctcac aattatgggc aggatggacc tatagaaggc 1560

aaaatttagc ctgctggcgg ttccccacac tgtaaaccaa agccctgaaa ggccatcata 1620

tcgttccgaa ggagaaggca aaaaacgcat cccaaggtgc tggaaggtca ccggcctgca 1680

gggtcaatct tgatgtcatt cgaggccttc cgaatttggc ttcctttctt ggctctgaaa 1740

tgaaatttag ttgtcatata cagatggtgt ctagcaatca cggctgcact cggtgtggct 1800

tggggtatgg tggcttgagc ttttccagct gctattggat tacacttgct ttgtgttgtc 1860

actgctgtcc ctctgctggg tccccgctgt cattaaggta tcgcagtccc cacctggtgt 1920

tctttctagc catctacagt acagtcatgc actcaaatta agattaagga aaaagatatt 1980

tttaagtgag taacttcatg cgcaataaaa aaaagacatg gctttttcta atgtggttta 2040

tctgtgattt aaaaaaaaaa aacacatgaa cttaacaata tttaaaacct gctacaatca 2100

gtgctgaaaa tagtattttc ccccttttct gcattttact attgtaaata tgttttctaa 2160

atcaaatact ttaaaagaag aaatgttgga tttataaatg ctatttttta ttttactttt 2220

ataataaaag tacaagcaca ctgtt 2245

<210>10

<211>966

<212>DNA

＜213〉Rattus norvegicus (norway rat)

<400>10

gagcctgagt tctcccagga gcagagtgag caggcgcagg agctacaaca tacaccacag 60

ggctgtgagg acagactctc gtgcctgtga gtgtcggtga gtcaaaccct ggggactgag 120

tccagtgcct cttctagcgt ccttggcatc atgggtcttc ggcttggcac caagatcact 180

gcgtgggact gctttccaaa ggatgtcatt gtccacaaaa gagagaaagc tggacacaca 240

tatgaaaatg gagaggacct gctgattggt gtacctagtg aagccacagt tcttgggact 300

atccagctaa tatgtgacct gatgattgca agctttgggg gaattttggt gtcagcttcc 360

tactccaacc cagaagcttc caccactttg atgtctggat acctgtttac tggatccttc 420

tgttttgcca tggctggaat tctctccatc atctctgaga aaatctcaac caaagccttt 480

gcactgagca gcctggcctc cagtgtagca agctctgttg ttgctatgat cggtctcttc 540

ttcgtcacct attgcctgat agtcctgggg actgcctttc cacactgtaa ctcagaaaag 600

aaattcctgc ccttattgtc ttatttggaa tcccatgatt ggaagaatga agacaagaac 660

tgtcgcctgg cttatgtcag tgctataagt actctgggga tcatgctcct cttcatcgtg 720

ctggaagttt tgctagctgt atacagttct atcttttggt ggaagcaagt ctactccaac 780

aagcctgggg gtgcattttt cttgcctcaa tcacaagatc atccctaact tgtcaaaagc 840

agtttgttgc agtaatggat atgaaataca gaagaagaaa gaagaaatct agtgcttgtg 900

gcagagcaca gctagacttt cctgaaatct caaccttggt agatcctaat aaacatctga 960

agaaaa 966

<210>11

<211>618

<212>DNA

＜213〉rat kind (Rattus sp)

<400>11

acacttctat gaagacaaga gcatttatta atttgaaagc atatcatctt taatacaaac 60

atttatagac tgcataacac attgcaatta aacatttctg taaaatagag tttatttatt 120

agtataacct tactgtataa tatagtatta ggcaagaatt gaaagaacgt aatgagagaa 180

cagggttgga tgatgtcaat gttgtcaaat ttgtggattt tacttttcac tatctctctg 240

tttggacttc tacaccaact ttaaaattcc aaatgtgaga cataaatgct tgtgtggaag 300

tatgccatca tcatcatcat tcaagcactg attggtgtta ttttaatgtg gctcagtcac 360

ttattgagat agaaggtggc ccactctgca gatattggag atcattagca tgagcatgcg 420

ggtgtgaaca ggagtgttgc tctctctaat ggggaattgt tcctcagggt acatctgact 480

gcagagtgaa gtctcagata catacctggg ttaagacagt cccatttaat atgtagagag 540

aaatacatct ttataacaca tcttttggat tgcttaaaaa tggaagtgac caacagctaa 600

tctggagtca gaatgagt 618

<210>12

<211>503

<212>DNA

＜213〉Rattus norvegicus (norway rat)

<400>12

cggccgctgc ggcgggagga gccccccagg aggaggacgg ggatccatgt gtctttcctg 60

gtgactagga tgtcgtcgga ggaggaccgg agcgcggagc agcagcagcc gccgccagca 120

ccccccgagg agcccggagc cccggccccg agccccgcag ccgcagacaa aagacctcgg 180

ggccggcctc gcaaagatgg cgcttcccct ttccagagag ccagaaagaa gcctcgaagt 240

agaggaaaat ctacagtgga agatgaggac agcacggatg gactggagac gacggagaca 300

gaaaatattg tggagacaga agtcaaagaa caatctgtgg aggaggacgc tgagacagaa 360

gtggagagca gcaagcagcc agcctcagct cttcagcggt ctgtgtctga ggactctgca 420

cactccctgg tctctgtcgg tgtagaagcc aaaatcagtg aacagctctg cgccttttgt 480

tactgtgggg agaaaagttc ctt 503

<210>13

<211>618

<212>DNA

＜213〉rat kind (Rattus sp)

<400>13

caaaattcta aaaagtttat ttgaaatcaa gccacccttc ccttaggctg cacggtagtt 60

cctcaaaggc agacaggtct gcagtgatgg taacaattca gcatgatcca gcttggggtc 120

cacaggaaca ggtgagaaca aaccctgcag gagctttcca ctgaaagcat cctggtataa 180

caggtactta cgtgagttct cattccggca gcttctattc taatgaactt gtaaatgttg 240

gaagtccaat ggcctttaaa ccatagcttt taccataatt gagaggaggt gacaggcagg 300

caggagccgc agaaaccatg atacatccac ttgtgttccc agaccccaga aatggtctta 360

catgacaata gttaagctca cagcaaaatc tgccatcagt gctccagctg tctagcatgt 420

gagaggtgac gtgacatcat atcagagctg agaccacgca gcttgtattt aacatgtggg 480

tgaggaaaga ggtcaaagta gagactataa acctgacgtt ggtgcactga ggcccacact 540

gtcaagtgca ctctcttagc tgtaagcagc agcacaccct ttgccaatgt ccacagcaga 600

gagcagacac agaccggt 618

Claims (11)

1, one group of buccal cavity white spot-related gene is characterized in that it expresses enhancing in buccal cavity white spot, and it has the nucleotide sequence shown in SEQID:1, SEQID:2.

2, one group of buccal cavity white spot-related gene is characterized in that it expresses enhancing in buccal cavity white spot, and it has the nucleotide sequence shown in SEQID:3, SEQID:4.

3, one group of buccal cavity white spot-related gene is characterized in that it expresses enhancing in buccal cavity white spot, and it has the nucleotide sequence shown in SEQID:5, SEQID:6.

4, one group of buccal cavity white spot-related gene is characterized in that it expresses enhancing in buccal cavity white spot, and it has the nucleotide sequence shown in SEQID:7, SEQID:8.

5, one group of buccal cavity white spot-related gene is characterized in that it expresses enhancing in buccal cavity white spot, and it has the nucleotide sequence shown in SEQID:9, SEQID:10.

6, one group of buccal cavity white spot-related gene is characterized in that it expresses enhancing in buccal cavity white spot, and it has the nucleotide sequence shown in SEQID:11, SEQID:12, SEQID:13.

7, genes involved according to claim 1 is characterized in that: the albumen of SEQID:1 genes encoding is that the albumen of chemokine (C-X-C motif) aglucon 9, SEQID:2 genes encoding is cyclin A2.

8, genes involved according to claim 2 is characterized in that: the albumen of SEQID:3 genes encoding is that the albumen that is similar to cyclin B2, SEQID:4 genes encoding is estrogen receptor 2.

9, genes involved according to claim 3 is characterized in that: the albumen of SEQID:5 genes encoding is that the albumen of Janus kinases 3, SEQID:6 genes encoding is chemokine C-X-C aglucon 10.

10, genes involved according to claim 4 is characterized in that: the albumen of SEQID:7 genes encoding is vehicle 1, and ATP-binding cassette, the albumen of B subfamily, SEQID:8 genes encoding are cell division cycle 2 homologue A.

11, genes involved according to claim 5 is characterized in that: the albumen of SEQID:9 genes encoding is that the albumen of nuclear factor KB p105 subunit, SEQID:10 genes encoding is membrane-spanning domain 4, A subfamily, 7.

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