CN1367246A - Nasopharyngeal carcinoma malignant transformation gene Tx gene DNA sequence and its determination method - Google Patents

Abstract

The present invention discloses gDNA sequence of nasopharyngeal carcinoma malignant transformation gene Tx. The Tx gene is expressed in the tumor cell of nasopharyngeal carcinoma and other epithelial source.

Description

Nasopharyngeal carcinoma malignant transformation gene Tx gene DNA sequence and measuring method thereof

The invention discloses the dna sequence dna of nasopharyngeal carcinoma malignant transformation gene Tx gene, belong to human malignancies molecular biology research field.

Recently domestic several research groups almost find the tumor cells expression immunoglobulin (Ig) of epithelial origin simultaneously.Only there is one piece of article to report abroad.Be the Kimoto report:, and further infer the existence that corresponding protein is arranged with the reorganization that has immunoglobulin (Ig) C district and tcr gene in the responsive nest-type PRC scientific discovery cancer epithelial cell.But up to now, yet there are no and be reported in the gene of being cloned into expressed immunoglobulin (Ig) in the cancer cells.The present invention has cloned one and immunoglobulin (Ig) kappa light chain gene height homologous Tx gene first from cancer cells, and proves that this gene has the vicious transformation function.Nasopharyngeal carcinoma malignant transformation gene Tx gene of the present invention is world's acquisition of cloning first, check order, and in U.S. gene database GenBank login (accession number: AF269073).

The objective of the invention is to pass through the nasopharyngeal carcinoma malignant transformation gene Tx of being cloned, inquire into cancer cells and express the mechanism of immunoglobulin (Ig) and the factor of regulation and control Tx genetic expression, express the biological significance of immunoglobulin (Ig) to set forth cancer cells, for the research of cell carcinogenesis mechanism provides a new thinking.

The method that realizes the object of the invention is at first to clone the gDNA of Tx gene.The gDNA of human nasopharyngeal carcinoma cell line CNE2 is broken into the fragment of 9-22kb, carrier λ Dash recombinates, transfection JB6 cell strain, the positive colony that picking forms on soft agar, the gDNA that extracts this positive colony is transfection JB6 cell strain again, after three times such screenings, the positive colony that is obtained is set up the gDNA library, and the tumor-necrosis factor glycoproteins Alu of personnel selection screens this library as probe, obtain the gDNA sequence that is about 16kb, i.e. a Tx gene.

Description of drawings: Fig. 1 is the cleavage map spectrogram of Tx gene.

Below to gene of the present invention clone and sequence measurement be described further.

The cloning process of Tx gene is: adopt the source of the CNE2 nasopharyngeal carcinoma cell system of Epstein-Barr virus feminine gender as clone's vicious transformation gene.Cultivate the CNE2 cell with the BEM that contains 7.5% foetal calf serum (FBS) (basic Eagel`s substratum), extract the gDNA (15 μ g) of this cell, calcium phosphate precipitation transfection to 2 * 10 ⁵Among the recipient cell JB6, transfection one day, and after cultivating three days, with 5 * 10 ⁴Individual cell or 2.4 * 10 ⁶Individual cell is resuspended in 0.33% agar that contains 5%FBS, place 100-mm or 60-mm culture dish to cultivate respectively 12 days, counting is greater than clone's number of 8 cells, and deduction negative control value (no DNA or do not have recipient cell DNA) is the positive colony number that the non-dependence of berthing is grown.The cell clone of picking maximum from agar as cells transfected, further carries out clonal expansion again.Extract these clones' DNA and purifying to carry out the transfection of next round.With the 16kb fragment among the DNA that extracts liposome method and pSV2 cotransfection JB6 recipient cell, the neomycin resistance cell selective grows among the 5%TMEM that contains G418 (500ug/ml), cultivated for 2 weeks, changed liquid once in per three days therebetween, picking 3.6 * 10 in 0.33% agar ⁴The resistant cell non-dependence grown cell that promptly obtains to berth.The resistant cell line in 3-4 generation of going down to posterity is resuspended in 0.33% agar that contains 10%FBS, places the 60-mm culture dish.After 14 days, counting is greater than the positive colony number of 8 cells.To do part digestion through the DNA of three-wheel cells transfected with Mbol, the fragment of separating 9-22kb wherein by sucrose gradient centrifugation, partly fill and lead up these fragments with the T4 ligase enzyme with dGTP and dATP, in addition with Xhol digestion λ dash vector arms (10ug), behind the purifying, partly fill and lead up end with dTTP and dCTP, both are recombinated, thereby (formed Sal 1 restriction enzyme site, help like this from carrier, separating and insert fragment) pack this recombinant chou with Gigapack Gold packaging extract (stratagene Inc.), selective conversion p2392 intestinal bacteria.This library capacity that do not increase is 8 * 10 ⁶Recombinant phage.Phage is after amplification, and this gDNA library titre is 1.4 * 10 ¹⁰Pfu/ml.

The Alu repeated sequence of personnel selection is as probe, and the screening phage library is chosen positive colony, and with its DNA of Sal 1 complete digestion, the insertion fragment of purifying 16-kb in gel is with it called after Tx gene (nasopharyngeal carcinoma deterioration transforming gene).

The method that the nasopharyngeal carcinoma malignant transformation gene Tx gene of being cloned carries out sequencing is:

The Tx gene is carried out enzyme with Sal I, Xho I, EcoR I respectively to be cut, obtain five endonuclease bamhis, be respectively Sal I/Xho I Tx1.3, Xho I/EcoRI Tx3.0, EcoRI/EcoRI Tx 1.0, EcoR I/EcoR I TX2.8 and EcoR I/Sal I Tx6.0.Restriction enzyme mapping as shown in Figure 1.

Method one: with the blank plasmid of PSP72 as carrier, respectively with the endonuclease bamhi subclone in this carrier, with recombinant plasmid dna as sequencing template.Initial primers is respectively carrier T7 Promoter and SP6 Promoter, from inserting the order-checking in opposite directions simultaneously of fragment 5 ' end and 3 ' end, is advanced into coincidence gradually.Method two: based on the known array of measuring, the design primer carries out PCR, and amplified production checks order, with the sequencing result of verification method one.Sequencing reaction: adopt two deoxidation cessation method.

Determine that through the aforesaid method order-checking Tx full length gene is 13kb.

Tx gene sequences are: 1 gtgtggaggg aaaggtgtga acccgggagg cggagcttgc agtgagccaa gatcgcacca 61 ctgcactcca gcctgggcga cagagcgagac tccgtctca aaaaaaataa ataaataaaa 121 taaaaaataa ttaggtactc taggcccagt gacctgtctc tgtactctgt aaattcaggt 181 cacctgctca gggctaatct gagagaaggt ctctcttcag ttgaattttg aaagacaatt 241 agcagttcac aagctaaccc aggtggacaa agatgttccc aagcagaggg agtgcttgtg 301 aaagctggag gccatagaaa aactctaagg agtgtaggga ggtgggagta atgtatggaa 361 ggggtggaga tggaaggtta agagagatac aaggctgcaa aaatggagct ggactcaaaa 421 gaaaatactg aaaaggtctt cagtgttgtt gatgagatta ctatggaaac actatggaac 481 actgggactc catggcagct ccaaagatgg catgcgcctg gtccagctca gtaagagctg 541 agctcttcct gtgctgtgaa aacagacaaa ccaaccaagt aaagtctact tttctactct 601 attagtcttc actttggttt cgtataccat ctggagctac atttcaaaat gcatttcaaa 661 gttatgagcc ttaagttgat atatatttag tctacctttt tttaaataac attgcagcaa 721 aggagaagat aaaatagtaa gacaacctgt aattattact cattgagaag ctgatgattt 781 ccataattac actaaatgaa gtttatcctt tgcaaaagcc cccccagccc accccaaaag 841 aaagtacaaa aaaactggcc atttttttta attgcttgtt tttctttgta attaacattc 901 agtctacttt ctaaaaaata aataaataat aagcagtcca gatgtggcaa gttgctaaag 961 aaaggaacca tcaggccata gacgtaaata tattctcttc ttggatttta ggtctcacct 1021 aagaaaataa acacatgcta tgtcagagaa gcctcagggt ttccacacct gctcgaaaag 1081 ggagttgagc ttcagcagct gacccaggac tctgttcccc tttggtgaga agggtttttg 1141 ttcagcaaga caatggagag ctctcactgt ggtggacgtt cggccaaggg accaaggtgg 1201 aaatcaaacg tgagtagaat ttaaactttg cttcctcagt tgtctgtgtc ttctgttccc 1261 tgtgtctatg aagtgatcta taaggtgact ctgcaatcag cctctgatat ccttcaggga 1321 aaagataaag ataagtctgt agtcaaactc gagaattgat tgcacatttt ctttgaagag 1381 caagcaagat tcagtcattg ggtgagaata acttgtctaa gtaatagctt cagaaatgtc 1441 ctggggaaca taacatgttc tggacagagc cttggtcaat tgtcagaaag ggagtttttg 1501 tataggaggg aagttaagag gaaccatgtg tgtacacttt tggccagggg accaagctgg 1561 agatcaaacg taagtacttt tttccactga ttcttcactg ttgctaatta gtttactttg 1621 tgttcctttg tgtggatttt cattagtcgg atgccaggga tctaacaaac ttcattccca 1681 ggttaggtac agaggagggg aaattgttcc acaggacgct agcttgtggc taatttttaa 1741 gatttctaaa tcaaaataac ttcattgggg gaaagaggct tgctgagctt tcagggaggt 1801 ttttgtaaag ggaaaagtta agacgaatca ctgtgattca ctttcggccc tgggaccaaa 1861 gtggatatca aacgtaagta catctgtctc aattattcgt gagattttag tgccattgta 1921 tcatttgtgc aagttttgtg atattttggt tgaataaacc tggtgaccca gaagtaaata 1981 gcaggacacc agaaaatgaa cttaaaaagc tgagcaaata gacgaatcat tgggtttgag 2041 aggagaatag gattcatggg ggaaatgggg aagaaatagc tagatttttc tctgaacaag 2101 cagcctatct catatgattg gcttcaagag aggtttttgt tgaggggaaa gggtgagatc 2161 cctcactgtg gctcactttc ggcggaggga ccaaggtgga gatcaaacgt aagtgcactt 2221 tcctaatgct ttttcttata aggttttaaa tttggagcgt ttttgtgttt gagatattag 2281 ctcaggtcaa ttccaaagag taccagattc tttcaaaaag tcagatgagt aagggataga 2341 aaattagttc atcttaagga acagccaagc gctagccagt taagtgaggc atctcaattg 2401 caagattttc tctgcatcgg tcaggttagt gatattaaca gcgaaaagag atttttgtta 2461 aggggaaagt aattaagtta acactgtgga tcaccttcgg ccaagggaca cgactggaga 2521 ttaaacgtaa gtaatttttc actattgtct tctgaaattt gggtctgatg gccagtattg 2581 acttttagag gcttaaatag gagtttggta aagattggta aatgagggca tttaagattt 2641 gccatgggtt gcaaaagtta aactcagctt caaaaatgga tttggagaaa aaaagattaa 2701 attgctctaa actgaatgac acaaagtaaa aaaaaaaagt gtaactaaaa aggaaccctt 2761 gtatttctaa ggagcaaaag taaatttatt tttgttcact cttgccaaat attgtattgg 2821 ttgttgctga ttatgcatga tacagaaaag tggaaaaata cattttttag tctttctccc 2881 ttttgtttga taaattattt tgtcagacaa caataaaaat caatagcacg ccctaagaaa 2941 aatcagggaa aagtgaagtg tacctatttg ctatgtagaa gaggcagctt acttgaaaat 3001 cagcagcaat gttgttttta gagtctgtaa taagtaataa actcaaaaag acacattcta 3061 taggaataag ggcttcacag atagagctca ttttttaaaa atccaatttg tacattagac 3121 taaacgtgaa attatctctt attgtaatgg tggaaaggtg gttattccca aaagctcaat 3181 ttcaaagaaa tgtgtttaaa tgaaaaaaag taaataattg cattttttaa tgaccgtggg 3241 tctgtgaaaa aaataggaaa tattttaaag agtatgttct ttcattatcc tctgttatta 3301 cttgtctaca tttttattct gccaagaagg ccgtggcacc gcgagctgta gacagagccg 3361 cggtctttct cgattgagtg gctttggtgg ccatgccacc gcgctcttgg ggcagccgcc 3421 ttgccgctag tggccgtggc caccctgtgt ctgcccgatt gatgctgccg tagccagctt 3481 tcctgatgca cagtgataca aataatgcca ctaagggaaa gagaacagaa acgtaatggg 3541 cgctgagctg ggaaaaccag ggagaagact gatttattag agatttcaga aataaaattc 3601 acattcatta tgatatctca ttagtgaaaa tttccattag gggattgtaa ataatttaaa 3661 gctttttttt ttcagtgcta tttaattatt tcaatatcct ctcatcaaat gtatttaaat 3721 aacaaaagct caaccaaaaa gaaagaaata tgtaattctt ttcagagtaa aaatcacacc 3781 catgacctgg ccactgaggg cttgatcaat tcactttgaa tttggcatta aataccatta 3841 aggtatatta actgatttta aaataagata tattcgtgac catgttttta actttcaaaa 3901 atgtagctgc cagtgtgtga ttttatttca gttgtacaaa atatctaaac ctatagcaat 3961 gtgattaata aaaacttaaa catattttcc agtaccttaa ttctgtgata ggaaaatttt 4021 aatctgagta ttttaatttc ataatctcta aaatagttta atgatttgtc attgtgttgc 4081 tgtcgtttac cccagctgat ctcaaaagtg atatttaagg agattatttt ggtctgcaac 4141 aacttgatag gactatttta gggccttttt aaagctctat taaaactaac ttacaacgat 4201 tcaaaactgt tttaaactat ttcaaaatga ttttagagcc ttttgaaaac tcttttaaac 4261 actttttaaa ctctattaaa actaataaga taacttgaaa taattttcat gtcaaataca 4321 ttaactgttt aatgtttaaa tgccagatg aaaaatgtaaa gctatcaaga attcacccag 4381 ataggagtat cttcatagca tgtttttccc tgcttatttt ccagtgatca cattattttg 4441 ctaccatggt tattttatac aattatctga aaaaaattag ttatgaagat taaaagagaa 4501 gaaaatatta aacataagag attcagtctt tcatgttgaa ctgcttggtt aacagtgaag 4561 ttagttttaa aaaaaaaaaa aactatttct gttatcagct gacttctccc tatctgttga 4621 cttctcccag caaaagattc ttattttaca ttttaactac tgctctccca cccaacgggt 4681 ggaatccccc agagggggat ttccaagagg ccacctggca gttgctgagg gtcagaagtg 4741 aagctagcca cttcctctta ggcaggtggc caagattaca gttgacctct cctggtatgg 4801 ctgaaaattg ctgcatatgg ttacaggcct tgaggccttt gggagggctt agagagttgc 4861 tggaacagtc agaaggtgga ggggctgaca ccacccaggc gcagaggcag ggctcagggc 4921 ctgctctgca gggaggtttt agcccagccc agccaaagta acccccggga gcctgttatc 4981 ccagcacagt cctggaagag gcacagggga aataaaagcg gacggaggct ttccttgact 5041 cagccgctgc ctggtcttct tcagacctgt tctgaattct aaactctgag ggggtcggat 5101 gacgtggcca ttctttgcct aaagcattga gtttactgca aggtcagaaa agcatgcaaa 5161 gccctcagaa tggctgcaaa gagctccaac aaaacaattt agaactttat taaggaatag 5221 ggggaagcta ggaagaaact caaaacatca agattttaaa tacgcttctt ggtctccttg 5281 ctataattat ctgggataag catgctgttt tctgtctgtc cctaacatgc cctgtgatta 5341 tccgcaaaca acacacccaa gggcagaact ttgttactta aacaccatcc tgtttgcttc 5401 tttcctcagg aactgtggct gcaccatctg tcttcatctt cccgccatct gatgagcagt 5461 tgaaatctgg aactgcctct gttgtgtgcc tgctgaataa cttctatccc agagaggcca 5521 aagtacagtg gaaggtggat aacgccctcc aatcgggtaa ctcccaggag agtgtcacag 5581 agcaggacag caaggacagc acctacagcc tcagcagcac cctgacgctg agcaaagcag 5641 actacgagaa acacaaagtc tacgcctgcg aagtcaccca tcagggcctg agctcgcccg 5701 tcacaaagag cttcaacagg ggagagtgtt agagggagaa gtgcccccac ctgctcctca 5761 gttccagcct gaccccctcc catcctttgg cctctgaccc tttttccaca ggggacctac 5821 ccctattgcg gtcctccagc tcatctttca cctcaccccc ctcctcctcc ttggctttaa 5881 ttatgctaat gttggaggag aatgaataaa taaagtgaat ctttgcacct gtggtttctc 5941 tctttcctca tttaataatt attatctgtt gttttaccaa ctactcaatt tctcttataa 6001 gggactaaat atgtagtcat cctaaggcgc ataaccattt ataaaaatca tccttcattc 6061 tattttaccc tatcatcctc tgcaagacag tcctccctca aacccacaag ccttctgtcc 6121 tcacagtccc ctgggccatg gtaggagaga cttgcttcct tgttttcccc tcctcagcaa 6181 gccctcatag tcctttttaa gggtgacagg tcttacagtc atatatcctt tgattcaatt 6241 ccctgagaat caaccaaagc aaatttttca aaagaagaaa cctgctataa agagaatcat 6301 tcattgcaac atgatataaa ataacaacac aataaaagca attaaataaa caaacaatag 6361 ggaaatgttt aagttcatca tggtacttag acttaatgga atgtcatgcc ttatttacat 6421 ttttaaacag gtactgaggg actcctgtct cgcaagggcc gtattgagta ctttccacaa 6481 cctaatttaa tccacactat actgtgagat taaaaacatt cattaaaatg ttgcaaaggt 6541 tctataaagc tgagagacaa atatattcta taactcagca atcccacttc tagatgactg 6601 agtgtcccca cccaccaaaa aactatgcaa gaatgttcaa agcagcttta tttacaaaag 6661 ccaaaaattg gaaatagccc gattgtccaa caatagaatg agttattaaa ctgtggtatg 6721 tttatacatt agaataccca atgaggagaa ttaacaagct acaactatac ctactcacac 6781 agatgaatct cataaaaata atgttacata agagaaactc aatgcaaaag atatgttctg 6841 tatgttttca tccatataaa gttcaaaacc aggtaaaaat aaagttagaa atttggatgg 6901 aaattactct tagctggggg tgggcgagtt agtgcctggg agaagacaag aaggggcttc 6961 tggggtcttg gtaatgttct gttcctcgtg tggggttgtg cagttatgat ctgtgcactg 7021 ttctgtatac acattatgct tcaaaataac ttcacataa agaacatctta tacccagtta 7081 atagatagaa gaggaataag taataggtca agaccacgca gctggtaagt gggggggcct 7141 gggatcaaat agctacctgc ctaatcctgc cctcttgagc cctgaatgag tctgcttcca 7201 gggctcaagg tgctcaacaa aacaacaggc ctgctatttt cctgcatctg tgccctgttt 7261 ggctagctag gagcacacat acatagaaat taaatgaaac agaccttcag caaggggaca 7321 gaggacagaa ttaaccttgc ccagacactg gaaacccatg tatgaccact cacatgtttg 7381 ggaaggggga agggcacatg taaatgagga ctcttcctca ttctatgggg cactctggct 7441 gccctctcag ctactcatcc atccaacaca cctttctaag tacctctctc tgcctacact 7501 ctgaaggggt tcaggagtaa ctaacacagc atcccttccc tcaaatgact gacaatccct 7561 ttgtcctgct ttgtttttct ttccagtcag tactgggaaa gtggggaagg acagtcatgg 7621 agaaactaca taaggaagca ccttgccctt ctgcctcttg agaatgttga tgagtatcaa 7681 atctttcaaa ctttggaggt ttgagtaggg gtgagactca gtaatgtccc ttccaatgac 7741 atgaacttgc tcactcatcc tgggggccaa attgaacaat caaaggcagg cataatccag 7801 ttatgaattc tttggaagaa gagtcttcta ttgtgagact tcactttggg tgggtattgg 7861 attcgttcac ttctgtccac gacgtccacc ttaacacagg aagtttttcc atacgagttg 7921 aggaacgaga accatgagta tttacccagt gtatttacac tgaaataaac ctttatccca 7981 gaaacgtctc cattagttca gttttaatcc agtatgactt tacaaacact cctacgccac 8041 ttttacctag taagtatata acgaccaccc ttatattttc ccatatcgat gagatctttt 8101 atcaacagtc aaagaacttt ttgatttgtt ttctgtggat ggtatactgg gtccttaaca 8161 tgaggaaccc ttaaatgggg gtcctttatt tttgaataca ggtgtgtctt gggtatgtac 8221 taacaagttg tcgtcgaaat aaacaacatc ggtttcgatc tttctcggtt gggtagggag 8281 ttatccgttg atcggattgt ttaacattat atagcccagt gtatggtaca ctaaggaaaa 8341 tacattgtaa caacttcact gttttaagta cggtatctta cgatactccg ttatttttcc 8401 ttgcttcaca actatgtctc ttgacctcac taagacttcc tgaaagatga ctcacttttt 8461 tcggttagac ttttatccct atctcttgtc taagaccaac ggtccccaat cccaccacct 8521 ctttcttctc atccgctttg atatttccct ctagaaacac tagtacccta tttagacata 8581 gaactaacgt caccatcaac gtccgtagat ctgtacacta ttttactgta tcttgacatg 8641 tgtgaataaa atagttacag tttaagaacc aaaattatag catgacatta atgcattctt 8701 cattggttgt cctctttgac ccacgtcctg tgtagtctgg agacacgaaa tataggacag 8761 aaacgatgaa agacacttag atattaataa aggtttatta aaaaaatttg aaaaaaaata 8821 cgacctagct cgttaagatc tctaggggaa gcttcctttc tggactacga aaagcacgcg 8881 cgtattttat ggaactatga cacggcctac tttcgccaag cgctgctcat ctacgttaat 8941 accaaagagg cggttcttag agaaacgtaa atagttcaca aaggaagtaa ctataaggct 9001 ctcgtagtta tacgttacga caaccctacc gttaaaaatg cggacaaaac gaaacgagct 9061 gtatttctat aggtagatgc tatagtctgg tgaagtaaag cgtatttagt ggttgagcaa 9121 cgggccattg ttgtcggtca aggtaacgtt cagactcggt tgtaccacta ctaagacgac 9181 gaactattta aaagtccata agcagtcggc attcagaact agaggaatgg agactaaaac 9241 gacgcgctca ccgtcgctgt accaaacaac aatataccgg aagtcgataa cggagagcct 9301 tacgtagcga gtcacaacta gactaattga accgactgcg gcggaacggg agcagataca 9361 taggtaactc gtaacggcgt taaagaaaac accactacag aagttcacct cgtagtccgt 9421 ctggggagg aataacgaaat taaaacgagt acattaaata ctcacagaag atgaactaag 9481 gagacgaccg gtcctaaaaa agcatcgcta gttcggtact tacattgcat tgccttaata 9541 gtgacaacta agagcgacag tctccgaaca caaacacagg acttttattg agttacaacc 9601 ggacatatcg aagtcactaa cgctaagcgg acagagacgg attaggtttg agaaatgggc 9661 aggaacccag ggacatcgtc attataggta acaaagaata tatttccaat cccccattta 9721 gggccgcgag tactgaagcg gaagaagggt aaagactagg agaagttttc cggtggacaa 9781 tgaccagcta aattcagttg gaaatggcga ctaagcacct tgtctatgag agaaggtagg 9841 aattggcctc cacccttata ggacgtaagg gcttgggtag ctgcttgaca aagttccgaa 9901 gaacctgcag cgaccgcacg caaggtgagg acttcacagt tcatgtagcg tttcagaggc 9961 gttaatgtgc gttctttttt ggcggtagtc cgccgaacca caagaaagtc aagaagttaa 10021 gcttataacc aatgcagacg tacacgatag acgcgggtat agtaggtcac cagcatcgtc 10081 agcaactaca agaggcgaag ctattgagac aacttaccga gaggtaaggt aagaggacac 10141 tgagccttca cgtaaatagt agaggtattt tgttttgggc ggcatcgctc aagtctattt 10201 tatttagggg cgctcacgct cctaacaata cattataacc caaattagta gatatacaaa 10261 acatgtctct cccgttcata gcaaaggtgg catgagcact attattaaaa cgtgccatag 10321 tcagtaaaga gcgtgtaacg tcttacccct aaacagaagt aatctgaata tttggaagta 10381 ccttataaac atacggctga gatatagata tggaagtaga tgtatttgtg gaagcactac 10441 agacgtacct ctgttctgtg gcctagacgt gttgtaacta ttgcgggtta gaaaaacgag 10501 tctgagattg agtaactatg agtaaatatt tgaggaacgt tacatacagc aaagtcgatt 10561 tgccatagtc gttacaaata catttctttg tcattctatt atgagttggg ctacaaactc 10621 atgccagtag tagactgtga tgtctgagac cgtagcgaca cttctgctgc gctttaagtc 10681 gtaaaagtgt tcgcaataga aaatgttttg gctagagtga gaggaaacta cgcttacggt 10741 cgcagtctgt agtatacgtc tatgagtgga cgtaggactt gggtaactgg aggttggggc 10801 attatcgcta cgcattacta cagctatcaa tgattgccca gaacaagcta attgacggcg 10861 tctttgagaa ggtccagtgg tcacgtcacg aactattgtc ctcagaaggg tcctaccgct 10921 tgttgttctt tgaccaaagg cagaagtgcc tgaagcaacg aaaggtcaaa tcgttatgcg 10981 aatgagggta ggctctattg tggaagcatt atgagtgcga cgagcaactc aaaactaaaa 11041 cgacaaagtt cgagttgtgc gtcaaaggga tgacaatcgc gttataggag caagaggacc 11101 agcgccgcaa actacataac gaccaaagaa agggcaagta ggtcgtcaag gtcgtgttag 11161 ctaccacaat ggttaagtac cttttccaga cgcagtttag gggtcagcag tacgtaacgg 11221 acgagacggc gaagtgcgtc acggactctc aattaaagcg agtgaagctt ggagagacaa 11281 atgactattc aaggtctagg aggaccgttg aacgtgttca ggctgttggg acttgctggt 11341 ccgcagaagc aagtagatag cctagcggtg tgagtgttgt tactcaccgt ctatatcgga 11401 ccaccaagtc cgccgcgtaa aaataacgac acaacgcgac attaagaaga taaagactac 11461 gacttagtta ctacagacgg tagaaagtaa ttagggactt gacaaccaat tatgcgaact 11521 cccacttacg cttattattt tttcctcgga catcgaggga ctactaaaac gaaaagtaca 11581 agtagcaagg aatttctgcg gcaaattgta cggctaacgg tccgaattta ctcagccaca 11641 cttagggtag tcgcaatggc aaagcgccac gaagaagtca tgcgatgccg tttacagtag 11701 ctgcaaaaat aggcctttga cgacagaccg aaaaaaacta aagtcttaat cggactgccc 11761 gttacgacgc ttcccgcaaa aggacgactc cacagtaact tgttcagggt acagccgttc 11821 gtattcgtgt gtcttatact tcgggcgacg gtctttttac gtaaggcacc aacagtatgg 11881 accagagaga gtagacgaag acgaaagcgg tggtagtaaa ggtcgaaaac actttcccta 11941 cgccgattgc atactttaag aagcagacaa agatgaccat aaccgtgttt ggactaaggt 12001 taaactcgtt ccgatacacg gtagagctat gagcaagaat tgagttgtct tctacgaaac 12061 acgtatgtcg gggagcaaat aataaataga ggagtcggtc ggcgacacga aagtcaccta 12121 aagcctattg tctttccggc cctttatggg tcggagcgaa acattgcctc atctgctttc 12181 actaacgcgg atgggcctat aatagcactc ctacgcagta gcggtaacga ggggtttatg 12241 ttttggttaa agtcggtcac ggagcaggta aaaaagctac ttgaggccgt gctagagcag 12301 ttttgagcgg tacatgaaaa gtagggcgag ttagtgctgt attacgtccg gaagtgcgaa 12361 gtatgcgccc agtatcaacc gtttcatggt ccgtaaaaaa gcgcagtggg tgtacgacat 12421 gacgtggacc cggtacattc gactgaaata ccggagcttt ggtggctcgg ccttgaagta 12481 ctttagggcc ctccatttgc ccgtaaagtc aagttccggc aacggcagtg acgtatttgg 12541 tagccctctc gtccgccatg cgtatgaaag cagcgctatc tactagcccc taagtcattg 12601 taagtgcggc cttcatttaa gtttgtccca agaccgcagc aagagcatga caaaaggggt 12661 ccggtcacga aatcgcaatt gaaggcctcg gtgtggccac gtttggagtc gttcgtccca 12721 caccttcatc ctgtaaaagt acagtccggt gaagaaaggc ctcgccccaa aacgatagtg 12781 caacacttga agacttcggc tactactgcg gctcggcatt aaacacggtg cgtagaaggg 12841 ggacaagctg tcgagagtgt agctagggcc atgcgacgtc ctattacagg ccacagtacg 12901 acggtggaag acgagacgcc gaaagacaaa gtccttaggt tctcgaaaat gacgaagccg 12961 gacacagtca agactgctac gtgcttacag cgccgcttta tagacccttg tctcgccgtt 13021 attcagcagt agggtacaaa ataggtcccg ctagtcgtct cacaattaga ggacgtacca 13081 aagtagcaat tggcctcact acagcgcaag gccgactgca agacgtcaca tacgtcataa 13141 aagctgttac gcgagccgaa gtagggaaca gtatctatgg tcgtttaggc ttccggtctg 13201 cccgtgtgac ttacgtaccg aaatactgca ttgtaggcaa accctacgct gacggtgccg 13261 gggcactaaa gagacggaag cgctcaaaac ttaccaagcg ccgccgtaag taggtaggta 13321 agccattgcg tctagcctac taatgccagg aacgccattt aggccgtaca tgtcctaagt 13381 aacaggacga gtttcaggta cggtagtttg acgaccaaaa gtaactacta cgccctggtc 13441 ggtagttgcg ggtggtggcc ttgctacggt aagacgaata gtcctttccg catttaaaga 13501 aagcaggtgc ctaattccgg catgaccaac cgttgctagt cattacgcta cttgacgcgt 13561 agcgaccgta gtggaaattt acggcagacc gcttctcacc actagtcaag gacac

Above-mentioned Tx gene and human normal immunoglobulin kappa light chain gene height homology, five J districts of J1-J5 of containing Ig κ, a C district, but the C district that lacks Ig κ, in addition, Tx also has recombination signal sequence (recombination signal sequence, RSS), N-segment, the relevant district of nuclear matrix (matrix associated region, sequence such as MAR), compare with the JC district gDNA of normal Ig kappa gene, the Tx gene exists disappearance or point mutation in some site, and the Tx gene is the unusual human normal immunoglobulin kappa light chain gene in a shortage V district.Utilize molecule clone technology also can directly clone the cDNA total length of Tx gene.

Above-mentioned Tx gene not only has expression in nasopharyngeal carcinoma cell system, and in the tumour cell of other epithelial origins expression is arranged also.By research gene order of nasopharyngeal carcinoma malignant transformation gene Tx and coded protein sequence, can determine the function of Tx gene and regulate and control the factor of this genetic expression, and illustrate the biological significance that cancer cells is expressed immunoglobulin (Ig).

Claims (5)

1 nasopharyngeal carcinoma malignant transformation gene Tx gene gDNA sequence total length 13kb with human normal immunoglobulin kappa light chain gene height homology, is characterized in that:

1) the Tx gene contains five J districts of J1-J5 of Ig κ, a C district, but lack

The C district of Ig κ, in addition, Tx also has recombination signal sequence

(recombination signal sequence, RSS), N-segment, nuclear base

The qualitative correlation district (matrix associated region, sequence such as MAR), with

The JC district gDNA of normal Ig kappa gene compares, and the Tx gene is in some position

Point exists disappearance or point mutation, and the Tx gene is the unusual of a shortage V district

Human normal immunoglobulin kappa light chain gene.

2) can directly clone the cDNA preface of Tx gene with the method for molecular cloning

Row.

2 according to the described Tx gene of claim 1, the method of nasopharyngeal carcinoma malignant transformation gene Tx gene of it is characterized in that cloning people is: with KB cell is the source of CNE2 as clone Tx gene, extract the gDNA of this clone, be broken into the fragment of 9-22kb, recombinate among the carrier λ Dash, transfection JB6 cell strain, the positive colony that picking forms on soft agar, extract the gDNA of this positive colony, it is interrupted once more, in the λ Dash carrier of recombinating, transfection JB6 clone, the positive colony that picking forms on soft agar, three screenings through such obtain a Tx gene that is about 13kb at last.

3 according to claim 1 obtained by sequencing gDNA Tx gene, identified by Tx gene sequences are: 1 gtgtggaggg aaaggtgtga acccgggagg cggagcttgc agtgagccaa gatcgcacca 61 ctgcactcca gcctgggcga cagagcgaga ctccgtctca aaaaaaataa ataaataaaa 121 taaaaaataa ttaggtactc taggcccagt gacctgtctc tgtactctgt aaattcaggt 181 cacctgctca gggctaatct gagagaaggt ctctcttcag ttgaattttg aaagacaatt 241 agcagttcac aagctaaccc aggtggacaa agatgttccc aagcagaggg agtgcttgtg 301 aaagctggag gccatagaaa aactctaagg agtgtaggga ggtgggagta atgtatggaa 361 ggggtggaga tggaaggtta agagagatac aaggctgcaa aaatggagct ggactcaaaa 421 gaaaatactg aaaaggtctt cagtgttgtt gatgagatta ctatggaaac actatggaac 481 actgggactc catggcagct ccaaagatgg catgcgcctg gtccagctca gtaagagctg 541 agctcttcct gtgctgtgaa aacagacaaa ccaaccaagt aaagtctact tttctactct 601 attagtcttc actttggttt cgtataccat ctggagctac atttcaaaat gcatttcaaa 661 gttatgagcc ttaagttgat atatatttag tctacctttt tttaaataac attgcagcaa 721 aggagaagat aaaatagtaa gacaacctgt aattattact cattgagaag ctgatgattt 781 ccataattac actaaatgaa gtttatcctt tgcaaaagcc cccccagccc accccaaaag 841 aaagtacaaa aaaactggcc atttttttta attgcttgtt tttctttgta attaacattc 901 agtctacttt ctaaaaaata aataaataat aagcagtcca gatgtggcaa gttgctaaag 961 aaaggaacca tcaggccata gacgtaaata tattctcttc ttggatttta ggtctcacct 1021 aagaaaataa acacatgcta tgtcagagaa gcctcagggt ttccacacct gctcgaaaag 1081 ggagttgagc ttcagcagct gacccaggac tctgttcccc tttggtgaga agggtttttg 1141 ttcagcaaga caatggagag ctctcactgt ggtggacgtt cggccaaggg accaaggtgg 1201 aaatcaaacg tgagtagaat ttaaactttg cttcctcagt tgtctgtgtc ttctgttccc 1261 tgtgtctatg aagtgatcta taaggtgact ctgcaatcag cctctgatat ccttcaggga 1321 aaagataaag ataagtctgt agtcaaactc gagaattgat tgcacatttt ctttgaagag 1381 caagcaagat tcagtcattg ggtgagaata acttgtctaa gtaatagctt cagaaatgtc 1441 ctggggaaca taacatgttc tggacagagc cttggtcaat tgtcagaaag ggagtttttg 1501 tataggaggg aagttaagag gaaccatgtg tgtacacttt tggccagggg accaagctgg 1561 agatcaaacg taagtacttt tttccactga ttcttcactg ttgctaatta gtttactttg 1621 tgttcctttg tgtggatttt cattagtcgg atgccaggga tctaacaaac ttcattccca 1681 ggttaggtac agaggagggg aaattgttcc acaggacgct agcttgtggc taatttttaa 1741 gatttctaaa tcaaaataac ttcattgggg gaaagaggct tgctgagctt tcagggaggt 1801 ttttgtaaag ggaaaagtta agacgaatca ctgtgattca ctttcggccc tgggaccaaa 1861 gtggatatca aacgtaagta catctgtctc aattattcgt gagattttag tgccattgta 1921 tcatttgtgc aagttttgtg atattttggt tgaataaacc tggtgaccca gaagtaaata 1981 gcaggacacc agaaaatgaa cttaaaaagc tgagcaaata gacgaatcat tgggtttgag 2041 aggagaatag gattcatggg ggaaatgggg aagaaatagc tagatttttc tctgaacaag 2101 cagcctatct catatgattg gcttcaagag aggtttttgt tgaggggaaa gggtgagatc 2161 cctcactgtg gctcactttc ggcggaggga ccaaggtgga gatcaaacgt aagtgcactt 2221 tcctaatgct ttttcttata aggttttaaa tttggagcgt ttttgtgttt gagatattag 2281 ctcaggtcaa ttccaaagag taccagattc tttcaaaaag tcagatgagt aagggataga 2341 aaattagttc atcttaagga acagccaagc gctagccagt taagtgaggc atctcaattg 2401 caagattttc tctgcatcgg tcaggttagt gatattaaca gcgaaaagag atttttgtta 2461 aggggaaagt aattaagtta acactgtgga tcaccttcgg ccaagggaca cgactggaga 2521 ttaaacgtaa gtaatttttc actattgtct tctgaaattt gggtctgatg gccagtattg 2581 acttttagag gcttaaatag gagtttggta aagattggta aatgagggca tttaagattt 2641 gccatgggtt gcaaaagtta aactcagctt caaaaatgga tttggagaaa aaaagattaa 2701 attgctctaa actgaatgac acaaagtaaa aaaaaaaagt gtaactaaaa aggaaccctt 2761 gtatttctaa ggagcaaaag taaatttatt tttgttcact cttgccaaat attgtattgg 2821 ttgttgctga ttatgcatga tacagaaaag tggaaaaata cattttttag tctttctccc 2881 ttttgtttga taaattattt tgtcagacaa caataaaaat caatagcacg ccctaagaaa 2941 aatcagggaa aagtgaagtg tacctatttg ctatgtagaa gaggcagctt acttgaaaat 3001 cagcagcaat gttgttttta gagtctgtaa taagtaataa actcaaaaag acacattcta 3061 taggaataag ggcttcacag atagagctca ttttttaaaa atccaatttg tacattagac 3121 taaacgtgaa attatctctt attgtaatgg tggaaaggtg gttattccca aaagctcaat 3181 ttcaaagaaa tgtgtttaaa tgaaaaaaag taaataattg cattttttaa tgaccgtggg 3241 tctgtgaaaa aaataggaaa tattttaaag agtatgttct ttcattatcc tctgttatta 3301 cttgtctaca tttttattct gccaagaagg ccgtggcacc gcgagctgta gacagagccg 3361 cggtctttct cgattgagtg gctttggtgg ccatgccacc gcgctcttgg ggcagccgcc 3421 ttgccgctag tggccgtggc caccctgtgt ctgcccgatt gatgctgccg tagccagctt 3481 tcctgatgca cagtgataca aataatgcca ctaagggaaa gagaacagaa acgtaatggg 3541 cgctgagctg ggaaaaccag ggagaagact gatttattag agatttcaga aataaaattc 3601 acattcatta tgatatctca ttagtgaaaa tttccattag gggattgtaa ataatttaaa 3661 gctttttttt ttcagtgcta tttaattatt tcaatatcct ctcatcaaat gtatttaaat 3721 aacaaaagct caaccaaaaa gaaagaaata tgtaattctt ttcagagtaa aaatcacacc 3781 catgacctgg ccactgaggg cttgatcaat tcactttgaa tttggcatta aataccatta 3841 aggtatatta actgatttta aaataagata tattcgtgac catgttttta actttcaaaa 3901 atgtagctgc cagtgtgtga ttttatttca gttgtacaaa atatctaaac ctatagcaat 3961 gtgattaata aaaacttaaa catattttcc agtaccttaa ttctgtgata ggaaaatttt 4021 aatctgagta ttttaatttc ataatctcta aaatagttta atgatttgtc attgtgttgc 4081 tgtcgtttac cccagctgat ctcaaaagtg atatttaagg agattatttt ggtctgcaac 4141 aacttgatag gactatttta gggccttttt aaagctctat taaaactaac ttacaacgat 4201 tcaaaactgt tttaaactat ttcaaaatga ttttagagcc ttttgaaaac tcttttaaac 4261 actttttaaa ctctattaaa actaataaga taacttgaaa taattttcat gtcaaataca 4321 ttaactgttt aatgtttaaa tgccagatga aaaatgtaaa gctatcaaga attcacccag 4381 ataggagtat cttcatagca tgtttttccc tgcttatttt ccagtgatca cattattttg 4441 ctaccatggt tattttatac aattatctga aaaaaattag ttatgaagat taaaagagaa 4501 gaaaatatta aacataagag attcagtctt tcatgttgaa ctgcttggtt aacagtgaag 4561 ttagttttaa aaaaaaaaaa aactatttct gttatcagct gacttctccc tatctgttga 4621 cttctcccag caaaagattc ttattttaca ttttaactac tgctctccca cccaacgggt 4681 ggaatccccc agagggggat ttccaagagg ccacctggca gttgctgagg gtcagaagtg 4741 aagctagcca cttcctctta ggcaggtggc caagattaca gttgacctct cctggtatgg 4801 ctgaaaattg ctgcatatgg ttacaggcct tgaggccttt gggagggctt agagagttgc 4861 tggaacagtc agaaggtgga ggggctgaca ccacccaggc gcagaggcag ggctcagggc 4921 ctgctctgca gggaggtttt agcccagccc agccaaagta acccccggga gcctgttatc 4981 ccagcacagt cctggaagag gcacagggga aataaaagcg gacggaggct ttccttgact 5041 cagccgctgc ctggtcttct tcagacctgt tctgaattct aaactctgag ggggtcggat 5101 gacgtggcca ttctttgcct aaagcattga gtttactgca aggtcagaaa agcatgcaaa 5161 gccctcagaa tggctgcaaa gagctccaac aaaacaattt agaactttat taaggaatag 5221 ggggaagcta ggaagaaact caaaacatca agattttaaa tacgcttctt ggtctccttg 5281 ctataattat ctgggataag catgctgttt tctgtctgtc cctaacatgc cctgtgatta 5341 tccgcaaaca acacacccaa gggcagaact ttgttactta aacaccatcc tgtttgcttc 5401 tttcctcagg aactgtggct gcaccatctg tcttcatctt cccgccatct gatgagcagt 5461 tgaaatctgg aactgcctct gttgtgtgcc tgctgaataa cttctatccc agagaggcca 5521 aagtacagtg gaaggtggat aacgccctcc aatcgggtaa ctcccaggag agtgtcacag 5581 agcaggacag caaggacagc acctacagcc tcagcagcac cctgacgctg agcaaagcag 5641 actacgagaa acacaaagtc tacgcctgcg aagtcaccca tcagggcctg agctcgcccg 5701 tcacaaagag cttcaacagg ggagagtgtt agagggagaa gtgcccccac ctgctcctca 5761 gttccagcct gaccccctcc catcctttgg cctctgaccc tttttccaca ggggacctac 5821 ccctattgcg gtcctccagc tcatctttca cctcaccccc ctcctcctcc ttggctttaa 5881 ttatgctaat gttggaggag aatgaataaa taaagtgaat ctttgcacct gtggtttctc 5941 tctttcctca tttaataatt attatctgtt gttttaccaa ctactcaatt tctcttataa 6001 gggactaaat atgtagtcat cctaaggcgc ataaccattt ataaaaatca tccttcattc 6061 tattttaccc tatcatcctc tgcaagacag tcctccctca aacccacaag ccttctgtcc 6121 tcacagtccc ctgggccatg gtaggagaga cttgcttcct tgttttcccc tcctcagcaa 6181 gccctcatag tcctttttaa gggtgacagg tcttacagtc atatatcctt tgattcaatt 6241 ccctgagaat caaccaaagc aaatttttca aaagaagaaa cctgctataa agagaatcat 6301 tcattgcaac atgatataaa ataacaacac aataaaagca attaaataaa caaacaatag 6361 ggaaatgttt aagttcatca tggtacttag acttaatgga atgtcatgcc ttatttacat 6421 ttttaaacag gtactgaggg actcctgtct cgcaagggcc gtattgagta ctttccacaa 6481 cctaatttaa tccacactat actgtgagat taaaaacatt cattaaaatg ttgcaaaggt 6541 tctataaagc tgagagacaa atatattcta taactcagca atcccacttc tagatgactg 6601 agtgtcccca cccaccaaa aaactatgcaa gaatgttcaa agcagcttta tttacaaaag 6661 ccaaaaattg gaaatagccc gattgtccaa caatagaatg agttattaaa ctgtggtatg 6721 tttatacatt agaataccca atgaggagaa ttaacaagct acaactatac ctactcacac 6781 agatgaatct cataaaaata atgttacata agagaaactc aatgcaaaag atatgttctg 6841 tatgttttca tccatataaa gttcaaaacc aggtaaaaat aaagttagaa atttggatgg 6901 aaattactct tagctggggg tgggcgagtt agtgcctggg agaagacaag aaggggcttc 6961 tggggtcttg gtaatgttct gttcctcgtg tggggttgtg cagttatgat ctgtgcactg 7021 ttctgtatac acattatgct tcaaaataac ttcacataaa gaacatctta tacccagtta 7081 atagatagaa gaggaataag taataggtca agaccacgca gctggtaagt gggggggcct 7141 gggatcaaat agctacctgc ctaatcctgc cctcttgagc cctgaatgag tctgcttcca 7201 gggctcaagg tgctcaacaa aacaacaggc ctgctatttt cctgcatctg tgccctgttt 7261 ggctagctag gagcacacat acatagaaat taaatgaaac agaccttcag caaggggaca 7321 gaggacagaa ttaaccttgc ccagacactg gaaacccatg tatgaccact cacatgtttg 7381 ggaaggggga agggcacatg taaatgagga ctcttcctca ttctatgggg cactctggct 7441 gccctctcag ctactcatcc atccaacaca cctttctaag tacctctctc tgcctacact 7501 ctgaaggggt tcaggagtaa ctaacacagc atcccttccc tcaaatgact gacaatccct 7561 ttgtcctgct ttgtttttct ttccagtcag tactgggaaa gtggggaagg acagtcatgg 7621 agaaactaca taaggaagca ccttgccctt ctgcctcttg agaatgttga tgagtatcaa 7681 atctttcaaa ctttggaggt ttgagtaggg gtgagactca gtaatgtccc ttccaatgac 7741 atgaacttgc tcactcatcc tgggggccaa attgaacaat caaaggcagg cataatccag 7801 ttatgaattc tttggaagaa gagtcttcta ttgtgagact tcactttggg tgggtattgg 7861 attcgttcac ttctgtccac gacgtccacc ttaacacagg aagtttttcc atacgagttg 7921 aggaacgaga accatgagta tttacccagt gtatttacac tgaaataaac ctttatccca 7981 gaaacgtctc cattagttca gttttaatcc agtatgactt tacaaacact cctacgccac 8041 ttttacctag taagtatata acgaccaccc ttatattttc ccatatcgat gagatctttt 8101 atcaacagtc aaagaacttt ttgatttgtt ttctgtggat ggtatactgg gtccttaaca 8161 tgaggaaccc ttaaatgggg gtcctttatt tttgaataca ggtgtgtctt gggtatgtac 8221 taacaagttg tcgtcgaaat aaacaacatc ggtttcgatc tttctcggtt gggtagggag 8281 ttatccgttg atcggattgt ttaacattat atagcccagt gtatggtaca ctaaggaaaa 8341 tacattgtaa caacttcact gttttaagta cggtatctta cgatactccg ttatttttcc 8401 ttgcttcaca actatgtctc ttgacctcac taagacttcc tgaaagatga ctcacttttt 8461 tcggttagac ttttatccct atctcttgtc taagaccaac ggtccccaat cccaccacct 8521 ctttcttctc atccgctttg atatttccct ctagaaacac tagtacccta tttagacata 858t gaactaacgt caccatcaac gtccgtagat ctgtacacta ttttactgta tcttgacatg 8641 tgtgaataaa atagttacag tttaagaacc aaaattatag catgacatta atgcattctt 8701 cattggttgt cctctttgac ccacgtcctg tgtagtctgg agacacgaaa tataggacag 8761 aaacgatgaa agacacttag atattaataa aggtttatta aaaaaatttg aaaaaaaata 8821 cgacctagct cgttaagatc tctaggggaa gcttcctttc tggactacga aaagcacgcg 8881 cgtattttat ggaactatga cacggcctac tttcgccaag cgctgctcat ctacgttaat 8941 accaaagagg cggttcttag agaaacgtaa atagttcaca aaggaagtaa ctataaggct 9001 ctcgtagtta tacgttacga caaccctacc gttaaaaatg cggacaaaac gaaacgagct 9061 gtatttctat aggtagatgc tatagtctgg tgaagtaaag cgtatttagt ggttgagcaa 9121 cgggccattg ttgtcggtca aggtaacgtt cagactcggt tgtaccacta ctaagacgac 9181 gaactattta aaagtccata agcagtcggc attcagaact agaggaatgg agactaaaac 9241 gacgcgctca ccgtcgctgt accaaacaac aatataccgg aagtcgataa cggagagcct 9301 tacgtagcga gtcacaacta gactaattga accgactgcg gcggaacggg agcagataca 9361 taggtaactc gtaacggcgt taaagaaaac accactacag aagttcacct cgtagtccgt 9421 ctggggagga ataacgaaat taaaacgagt acattaaata ctcacagaag atgaactaag 9481 gagacgaccg gtcctaaaaa agcatcgcta gttcggtact tacattgcat tgccttaata 9541 gtgacaacta agagcgacag tctccgaaca caaacacagg acttttattg agttacaacc 9601 ggacatatcg aagtcactaa cgctaagcgg acagagacgg attaggtttg agaaatgggc 9661 aggaacccag ggacatcgtc attataggta acaaagaata tatttccaat cccccattta 9721 gggccgcgag tactgaagcg gaagaagggt aaagactagg agaagttttc cggtggacaa 9781 tgaccagcta aattcagttg gaaatggcga ctaagcacct tgtctatgag agaaggtagg 9841 aattggcctc cacccttata ggacgtaagg gcttgggtag ctgcttgaca aagttccgaa 9901 gaacctgcag cgaccgcacg caaggtgagg acttcacagt tcatgtagcg tttcagaggc 9961 gttaatgtgc gttctttttt ggcggtagtc cgccgaacca caagaaagtc aagaagttaa 10021 gcttataacc aatgcagacg tacacgatag acgcgggtat agtaggtcac cagcatcgtc 10081 agcaactaca agaggcgaag ctattgagac aacttaccga gaggtaaggt aagaggacac 10141 tgagccttca cgtaaatagt agaggtattt tgttttgggc ggcatcgctc aagtctattt 10201 tatttagggg cgctcacgct cctaacaata cattataacc caaattagta gatatacaaa 10261 acatgtctct cccgttcata gcaaaggtgg catgagcact attattaaaa cgtgccatag 10321 tcagtaaaga gcgtgtaacg tcttacccct aaacagaagt aatctgaata tttggaagta 10381 ccttataaac atacggctga gatatagata tggaagtaga tgtatttgtg gaagcactac 10441 agacgtacct ctgttctgtg gcctagacgt gttgtaacta ttgcgggtta gaaaaacgag 10501 tctgagattg agtaactatg agtaaatatt tgaggaacgt tacatacagc aaagtcgatt 10561 tgccatagtc gttacaaata catttctttg tcattctatt atgagttggg ctacaaactc 10621 atgccagtag tagactgtga tgtctgagac cgtagcgaca cttctgctgc gctttaagtc 10681 gtaaaagtgt tcgcaataga aaatgttttg gctagagtga gaggaaacta cgcttacggt 10741 cgcagtctgt agtatacgtc tatgagtgga cgtaggactt gggtaactgg aggttggggc 10801 attatcgcta cgcattacta cagctatcaa tgattgccca gaacaagcta attgacggcg 10861 tctttgagaa ggtccagtgg tcacgtcacg aactattgtc ctcagaaggg tcctaccgct 10921 tgttgttctt tgaccaaagg cagaagtgcc tgaagcaacg aaaggtcaaa tcgttatgcg 10981 aatgagggta ggctctattg tggaagcatt atgagtgcga cgagcaactc aaaactaaaa 11041 cgacaaagtt cgagttgtgc gtcaaaggga tgacaatcgc gttataggag caagaggacc 11101 agcgccgcaa actacataac gaccaaagaa agggcaagta ggtcgtcaag gtcgtgttag 11161 ctaccacaat ggttaagtac cttttccaga cgcagtttag gggtcagcag tacgtaacgg 11221 acgagacggc gaagtgcgtc acggactctc aattaaagcg agtgaagctt ggagagacaa 11281 atgactattc aaggtctagg aggaccgttg aacgtgttca ggctgttggg acttgctggt 11341 ccgcagaagc aagtagatag cctagcggtg tgagtgttgt tactcaccgt ctatatcgga 11401 ccaccaagtc cgccgcgtaa aaataacgac acaacgcgac attaagaaga taaagactac 11461 gacttagtta ctacagacgg tagaaagtaa ttagggactt gacaaccaat tatgcgaact 11521 cccacttacg cttattattt tttcctcgga catcgaggga ctactaaaac gaaaagtaca 11581 agtagcaagg aatttctgcg gcaaattgta cggctaacgg tccgaattta ctcagccaca 11641 cttagggtag tcgcaatggc aaagcgccac gaagaagtca tgcgatgccg tttacagtag 11701 ctgcaaaaat aggcctttga cgacagaccg aaaaaaacta aagtcttaat cggactgccc 11761 gttacgacgc ttcccgcaaa aggacgactc cacagtaact tgttcagggt acagccgttc 11821 gtattcgtgt gtcttatact tcgggcgacg gtctttttac gtaaggcacc aacagtatgg 11881 accagagaga gtagacgaag acgaaagcgg tggtagtaaa ggtcgaaaac actttcccta 11941 cgccgattgc atactttaag aagcagacaa agatgaccat aaccgtgttt ggactaaggt 12001 taaactcgtt ccgatacacg gtagagctat gagcaagaat tgagttgtct tctacgaaac 12061 acgtatgtcg gggagcaaat aataaataga ggagtcggtc ggcgacacga aagtcaccta 12121 aagcctattg tctttccggc cctttatggg tcggagcgaa acattgcctc atctgctttc 12181 actaacgcgg atgggcctat aatagcactc ctacgcagta gcggtaacga ggggtttatg 12241 ttttggttaa agtcggtcac ggagcaggta aaaaagctac ttgaggccgt gctagagcag 12301 ttttgagcgg tacatgaaaa gtagggcgag ttagtgctgt attacgtccg gaagtgcgaa 12361 gtatgcgccc agtatcaacc gtttcatggt ccgtaaaaaa gcgcagtggg tgtacgacat 12421 gacgtggacc cggtacattc gactgaaata ccggagcttt ggtggctcgg ccttgaagta 12481 ctttagggcc ctccatttgc ccgtaaagtc aagttccggc aacggcagtg acgtatttgg 12541 tagccctctc gtccgccatg cgtatgaaag cagcgctatc tactagcccc taagtcattg 12601 taagtgcggc cttcatttaa gtttgtccca agaccgcagc aagagcatga caaaaggggt 12661 ccggtcacga aatcgcaatt gaaggcctcg gtgtggccac gtttggagtc gttcgtccca 12721 caccttcatc ctgtaaaagt acagtccggt gaagaaaggc ctcgccccaa aacgatagtg 12781 caacacttga agacttcggc tactactgcg gctcggcatt aaacacggtg cgtagaaggg 12841 ggacaagctg tcgagagtgt agctagggcc atgcgacgtc ctattacagg ccacagtacg 12901 acggtggaag acgagacgcc gaaagacaaa gtccttaggt tctcgaaaat gacgaagccg 12961 gacacagtca agactgctac gtgcttacag cgccgcttta tagacccttg tctcgccgtt 13021 attcagcagt agggtacaaa ataggtcccg ctagtcgtct cacaattaga ggacgtacca 13081 aagtagcaat tggcctcact acagcgcaag gccgactgca agacgtcaca tacgtcataa 13141 aagctgttac gcgagccgaa gtagggaaca gtatctatgg tcgtttaggc ttccggtctg 13201 cccgtgtgac ttacgtaccg aaatactgca ttgtaggcaa accctacgct gacggtgccg 13261 gggcactaaa gagacggaag cgctcaaaac ttaccaagcg ccgccgtaag taggtaggta 13321 agccattgcg tctagcctac taatgccagg aacgccattt aggccgtaca tgtcctaagt 13381 aacaggacga gtttcaggta cggtagtttg acgaccaaaa gtaactacta cgccctggtc 13441 ggtagttgcg ggtggtggcc ttgctacggt aagacgaata gtcctttccg catttaaaga 13501 aagcaggtgc ctaattccgg catgaccaac cgttgctagt cattacgcta cttgacgcgt 13561 agcgaccgta gtggaaattt acggcagacc gcttctcacc actagtcaag gacac ...

4 according to claim 1,3 described Tx genes, it is characterized in that the Tx gene not only has expression in nasopharyngeal carcinoma cell system, and in the tumour cell of other epithelial origins expression are arranged also.

5 according to right 1,3, clones the cDNA total length of Tx gene, it is characterized in that can be used for studying the factor of vicious transformation function, regulating and expressing of this sequence and the function of coded protein each side.

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