CN104561303A - 人肿瘤预测性生物标志组合物及其应用 - Google Patents
人肿瘤预测性生物标志组合物及其应用 Download PDFInfo
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Abstract
人肿瘤预测性生物标志组合物及其应用，包括BRAF、BRCA1、BRCA2、EGFR、FBXW7、H-RAS、IDH1、K-RAS、N-RAS、PIK3CA、PTEN和TP53基因中的至少两个。为中国8大常见肿瘤设定了模块式特定基因组合突变检测方案，为临床有效选择靶向抑制剂和化疗药物提供最新和有效筛选方案，以达到针对特定肿瘤的个体化治疗。本发明提供的方法包括2、4、6和12个基因组合图谱的检测方案和相应的数据分析，可快速全面地评估肿瘤细胞相关信号通路的功能变化以及对治疗方案的可能反应。此发明采用的突变检测方法比目前应用中的方法检测效率更高，检测结果更准确，应用范围广泛，可覆盖80％左右的癌症病例。
Description
技术领域
本发明属于肿瘤基因检测技术领域，具体涉及人肿瘤预测性生物标志组合物及其应用。
背景技术
人类癌症通过基因组畸变的积累而发展，如基因突变，使细胞的病理生理改变发生，如无限生长和转移。越来越多的证据显示人类癌症是一复杂和多样性疾病，患者可能表现出类似的症状，并具有相同的病理改变，但可能由完全不同的基因变化而造成。正因为这样的异质性，某一特定药物或疗法往往只对一部分肿瘤病人有效。因此，不同类型癌症晚期患者对目前可用药物的反应率差别很大，从10％至90％不等。许多较新的生物治疗方法，特别是所谓的靶向治疗(targeted therapy)，也仅仅在小部分具有特异基因突变的患者身上显示有疗效。因此，能前瞻性的确定患者有否可能对某特定治疗起反应的方法具有巨大的临床应用价值，可最大幅度的提高肿瘤的治愈率。
预测性生物标志物，如基因突变(gene mutation)和基因表达(gene expression)等，可以提供癌症患者是否可能从某特定治疗(如靶向药物)中获益的前期信息，从而用来指导治疗方法的选择。如病人的标志物状态表示对某特定治疗可能有效，则此病人为接受该治疗的候选者。相反，如标志物测定暗示对某一特定治疗有耐药性，则应寻找或获取可能更有益的替代疗法。总之，这些标志物的测定可以帮助临床医生做出明智的决定，以避免不必要的医疗费用，可能无效的治疗以及毒性，提高患者的生活质量。这些标志物也可用于疗效评估，帮助确定最佳药物剂量和毒性预测。本发明即为适合这样的需求而产生。
目前肿瘤基因治疗如靶向治疗极具潜力，有比传统化疗更大的优越性。而随着越来越多靶向药物的问世和应用或进入临床试验，包括新药审批，基于分子基因检测的生物标志物如基因突变在预测靶向治疗疗效中的作用越来越受重视。目前许多西方国家，如美国，正投入大量的人力和物力寻找这样的预测标志物，目前已有少数用于临床指导干预，包括中国医院。本发明者之一(毛建华博士)在《科学》等杂志发表的新基因路线图开发了一个预测新靶向治疗的方向，如针对FBXW7基因突变的成果已初步取得美国临床运用成功。
发明内容
解决的技术问题：本发明提供一种人肿瘤预测性生物标志组合物及其应用，可用于制备肿瘤预测或筛查试剂盒。
技术方案：人肿瘤预测性生物标志组合物，包括BRAF、BRCA1、BRCA2、EGFR、FBXW7、H-RAS、IDH1、K-RAS、N-RAS、PIK3CA、PTEN和TP53基因中的至少两个。
优选的方案，组合物中含有2、4、6或12个所述基因。
上述人肿瘤预测性生物标志组合物在制备肿瘤预测或筛查试剂盒中的应用。
上述试剂盒中还包括组合物所含基因的对应引物。
一种人肿瘤诊断试剂盒，含有检测BRAF、BRCA1、BRCA2、EGFR、FBXW7、H-RAS、IDH1、K-RAS、N-RAS、PIK3CA、PTEN和TP53中至少两个基因的引物。
优选的方案，含有2、4、6或12个所述基因的引物。
检测BRAF的引物序列如SEQ ID NO.1～2所示；检测BRCA1的引物序列如SEQ IDNO.4～11所示；检测BRCA2的引物序列如SEQ ID NO.13～18所示；检测EGFR的引物序列如SEQ ID NO.20～27所示；检测FBXW7的引物序列如SEQ ID NO.29～32所示；检测H-RAS的引物序列如SEQ ID NO.34～37所示；检测IDH1的引物序列为如SEQ ID NO.39～40所示；检测K-RAS的引物序列如SEQ ID NO.42～45所示；检测N-RAS的引物序列如SEQ IDNO.47～50所示；检测PIK3CA的引物序列如SEQ ID NO.52～55所示；检测PTEN的引物序列如SEQ ID NO.57～62所示；检测TP53的引物序列如SEQ ID NO.64～71所示。
所述试剂盒中还含有其它必要试剂：dNTPs、MgCl2、反应缓冲液和Taq DNA聚合酶。
有益效果：本发明通过对十二个基因突变图谱的检测达到对一系列靶向药物治疗进行系统性优化选择。根据此十二个基因在每个肿瘤中的累积突变率(图3)，其在八大恶性肿瘤的应用可覆盖80％左右的癌症病例。通过该试剂盒可以对治病样本中以上各基因进行扩增与纯化测序，并与相应的突变图谱进行检查对比，进而对样本中的肿瘤进行诊断、预测、检测或筛查，系统分析肿瘤通路中各成员的功能状态，包括通路之间和非通路之间的关系；增加基因的数目可增加预测的精确度；确定病人是否需要接受通路中下游基因的靶向治疗，或协同靶向治疗。
附图说明
图1为EGFR/RAS分子信息通路图；
图2为不同组合的检测基因突变图谱及其在中国八大常见肿瘤中的应用图，其中百分比表示十二个基因在每个肿瘤中的突变率；
图3为本发明十二个基因在每个肿瘤中的累积突变率图。
具体实施方式
下面的实施例可使本专业技术人员更全面地理解本发明，但不以任何方式限制本发明。
本发明不需要从肿瘤病人再次取样，而可以直接用现有病理检查的石蜡样本、肿瘤活检或血液标本。
从石蜡切片中提取高质量肿瘤样本DNA:先将待测样本去蜡，后将样本用蛋白酶K(Protinase K)消化，最后将DNA提取用于检测基因突变。肿瘤活检或血液标本DNA的提取类似于石蜡切片标本，除了不需去蜡步骤。对于血液标本，首先将肿瘤细胞从血液中分离出来。
本发明设计和优化了检测位于12个基因序列中突变热点的特定PCR引物。先应用PCR对相应的短基因片段进行扩充，然后对PCR产物进行纯化，最后应用DNA测序仪器来检测相应的基因是否有突变。本发明也开发了用第二代测序仪的方法来检测12个基因的突变，先用本发明设计的针对12个基因的DNAbaits将基因从总DNA中提取出来，然后建库测序。本发明还在在开发应用其它快速、精确和价廉的方法来检测12基因的突变图谱。
实施例1
通过检查单个基因IDH1的突变图谱决定肺癌、前列腺癌或其它癌症病人是否应接受IDH1靶向治疗(AGI-5198等)。
试剂盒组成：标准96孔PCR板，12个基因的引物(也可选择2、4、或6个基因的引物)，dNTPs、MgCl2、反应缓冲液和Taq DNA聚合酶。
用上述方法将DNA从病人肿瘤样本中提取出来。将病人肿瘤DNA(20～100ng)按照1：1容积比例加入到本发明设计好的96孔板试剂盒，然后将96孔板放入PCR仪对IDH1进行扩增，用Qiagen或其它PCR纯化试剂将PCR产物纯化，最后用DNA测序仪对IDH1基因进行突变检测。若病人肿瘤组织中含有IDH1突变，则IDH1的靶向治疗对此病人有效。
实施例2
通过检测EGFR和K-RAS的突变图谱决定肺癌病人是否应接受厄洛替尼(特罗凯)，吉非替尼(易瑞沙)或西妥昔单抗(爱必妥)的治疗[均为EGFR靶向治疗]。
用上述方法将DNA从病人肿瘤样本中提取出来。将病人肿瘤DNA(20～100ng)按照1：1容积比例加入到本专利设计的96孔板试剂盒，然后将96孔板放入PCR仪对EGFR和K-RAS进行扩增，用Qiagen或其它PCR纯化试剂将PCR产物纯化，最后用DNA测序仪对EGFR和K-RAS基因进行突变检测。若病人肿瘤组织中含有EGFR突变，但没有K-RAS突变，则EGFR的靶向治疗对此病人有效。
实施例3
通过检查BRAF1和K-RAS的突变图谱决定肠癌、肺癌或其它癌症病人是否应接受BRAF靶向治疗(如Vemurafenib或Dabrafenib)。
用上述方法将DNA从病人肿瘤样本中提取出来。将病人肿瘤DNA(20～100ng)按照1：1容积比例加入到本专利设计好96孔板试剂盒，然后将96孔板放入PCR仪对BRAF1和K-RAS进行扩增，用Qiagen或其它PCR纯化试剂将PCR产物纯化，最后用DNA测序仪对BRAF1和K-RAS进行突变检测。若病人肿瘤中含有BRAF1突变，但没有K-RAS突变，则BRAF1的靶向治疗对此病人有效.
实施例4
通过检查BRCA1和BRCA2的突变图谱决定乳癌、肺癌或其它癌症病人是否应接受PARP1靶向治疗(Inipaib等)。
用上述方法将DNA从病人肿瘤样本中提取出来。将病人肿瘤DNA(20～100ng)按照1：1容积比例加入到本专利设计好96孔板试剂盒，然后将96孔板放入PCR机对BRCA1和BRCA2进行扩增后，用Qiagen或其它PCR纯化试剂将PCR产物纯化，最后用DNA测序仪对BRCA1和BRCA2进行突变检测。若病人肿瘤组织中含有BRCA1突变，或有BRCA2突变，PARP1的靶向治疗对此病人有效。
实施例5
通过检测EGFR、K-RAS、PTEN和FBXW7的突变图谱决定肠癌病人是否应接受EGFR靶向治疗，如厄洛替尼(特罗凯)、吉非替尼(易瑞沙)或西妥昔单抗(爱必妥)或mTOR靶向治疗，如雷帕霉素或西罗莫司，或EGFR和mTOR的组合靶向治疗。
按上述方法将DNA从病人肿瘤样本中提取出来。将病人肿瘤DNA(20～100ng)按照1：1容积比例加入到本专利设计好96孔板试剂盒，然后将96孔放入PCR机对EGFR，K-RAS，PTEN和FBXW7进行扩增，用Qiagen或其它PCR纯化试剂将PCR产物纯化，最后用DNA测序仪对以上进行突变检测。若病人肿瘤组织中含有EGFR突变，但没有其它基因的突变，则EGFR的靶向治疗对此病人有效。若病人肿瘤组织中含有FBXW7和/或PTEN突变，实施以下方案：如没有其它基因的突变，采用mTOR靶向治疗，对此病人有效。若病人肿瘤中含有所有四个基因的突变，采用EGFR和mTOR协同靶向治疗，可以产生更好的治疗效果。
实施例6
通过检测EGFR、K－RAS、PI3K、PTEN、TP53和FBXW7的突变图决定肺癌或其它癌症病人是否应接受接受厄洛替尼(特罗凯)，吉非替尼(易瑞沙)或西妥昔单抗(爱必妥)的治疗[均为EGFR靶向治疗]。决定是否应实施雷帕霉素或西罗莫司[mTOR靶向治疗]，或EGFR和mTOR组合靶向治疗。与实施例5类似，增添TP53和PI3K的突变检测可以帮助决定应用单一或组合靶向治疗，如果此两基因均有突变，组合靶向应为首选。
实施例7
按照上面描述的方法将DNA从病人肿瘤样本中提取出来。通过检测所有的12个基因的突变图谱(图2)，决定癌症病人是否应接受组合治疗，如EGFR和mTOR联合靶向治疗。此实施是综合了实施例1-6。
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<213> 人工序列
<400> 9
tacttgtctg ttcatttggc ttgt 24
<210> 10
<211> 24
<212> DNA
<213> 人工序列
<400> 10
taaatatgac gtgtctgctc cact 24
<210> 11
<211> 24
<212> DNA
<213> 人工序列
<400> 11
acttatttat gtggttggga tgga 24
<210> 12
<211> 7224
<212> DNA
<213> 人工序列
<400> 12
gtaccttgat ttcgtattct gagaggctgc tgcttagcgg tagccccttg gtttccgtgg 60
caacggaaaa gcgcgggaat tacagataaa ttaaaactgc gactgcgcgg cgtgagctcg 120
ctgagacttc ctggacgggg gacaggctgt ggggtttctc agataactgg gcccctgcgc 180
tcaggaggcc ttcaccctct gctctgggta aagttcattg gaacagaaag aaatggattt 240
atctgctctt cgcgttgaag aagtacaaaa tgtcattaat gctatgcaga aaatcttaga 300
gtgtcccatc tgtctggagt tgatcaagga acctgtctcc acaaagtgtg accacatatt 360
ttgcaaattt tgcatgctga aacttctcaa ccagaagaaa gggccttcac agtgtccttt 420
atgtaagaat gatataacca aaaggagcct acaagaaagt acgagattta gtcaacttgt 480
tgaagagcta ttgaaaatca tttgtgcttt tcagcttgac acaggtttgg agtatgcaaa 540
cagctataat tttgcaaaaa aggaaaataa ctctcctgaa catctaaaag atgaagtttc 600
tatcatccaa agtatgggct acagaaaccg tgccaaaaga cttctacaga gtgaacccga 660
aaatccttcc ttgcaggaaa ccagtctcag tgtccaactc tctaaccttg gaactgtgag 720
aactctgagg acaaagcagc ggatacaacc tcaaaagacg tctgtctaca ttgaattggg 780
atctgattct tctgaagata ccgttaataa ggcaacttat tgcagtgtgg gagatcaaga 840
attgttacaa atcacccctc aaggaaccag ggatgaaatc agtttggatt ctgcaaaaaa 900
ggctgcttgt gaattttctg agacggatgt aacaaatact gaacatcatc aacccagtaa 960
taatgatttg aacaccactg agaagcgtgc agctgagagg catccagaaa agtatcaggg 1020
tagttctgtt tcaaacttgc atgtggagcc atgtggcaca aatactcatg ccagctcatt 1080
acagcatgag aacagcagtt tattactcac taaagacaga atgaatgtag aaaaggctga 1140
attctgtaat aaaagcaaac agcctggctt agcaaggagc caacataaca gatgggctgg 1200
aagtaaggaa acatgtaatg ataggcggac tcccagcaca gaaaaaaagg tagatctgaa 1260
tgctgatccc ctgtgtgaga gaaaagaatg gaataagcag aaactgccat gctcagagaa 1320
tcctagagat actgaagatg ttccttggat aacactaaat agcagcattc agaaagttaa 1380
tgagtggttt tccagaagtg atgaactgtt aggttctgat gactcacatg atggggagtc 1440
tgaatcaaat gccaaagtag ctgatgtatt ggacgttcta aatgaggtag atgaatattc 1500
tggttcttca gagaaaatag acttactggc cagtgatcct catgaggctt taatatgtaa 1560
aagtgaaaga gttcactcca aatcagtaga gagtaatatt gaagacaaaa tatttgggaa 1620
aacctatcgg aagaaggcaa gcctccccaa cttaagccat gtaactgaaa atctaattat 1680
aggagcattt gttactgagc cacagataat acaagagcgt cccctcacaa ataaattaaa 1740
gcgtaaaagg agacctacat caggccttca tcctgaggat tttatcaaga aagcagattt 1800
ggcagttcaa aagactcctg aaatgataaa tcagggaact aaccaaacgg agcagaatgg 1860
tcaagtgatg aatattacta atagtggtca tgagaataaa acaaaaggtg attctattca 1920
gaatgagaaa aatcctaacc caatagaatc actcgaaaaa gaatctgctt tcaaaacgaa 1980
agctgaacct ataagcagca gtataagcaa tatggaactc gaattaaata tccacaattc 2040
aaaagcacct aaaaagaata ggctgaggag gaagtcttct accaggcata ttcatgcgct 2100
tgaactagta gtcagtagaa atctaagccc acctaattgt actgaattgc aaattgatag 2160
ttgttctagc agtgaagaga taaagaaaaa aaagtacaac caaatgccag tcaggcacag 2220
cagaaaccta caactcatgg aaggtaaaga acctgcaact ggagccaaga agagtaacaa 2280
gccaaatgaa cagacaagta aaagacatga cagcgatact ttcccagagc tgaagttaac 2340
aaatgcacct ggttctttta ctaagtgttc aaataccagt gaacttaaag aatttgtcaa 2400
tcctagcctt ccaagagaag aaaaagaaga gaaactagaa acagttaaag tgtctaataa 2460
tgctgaagac cccaaagatc tcatgttaag tggagaaagg gttttgcaaa ctgaaagatc 2520
tgtagagagt agcagtattt cattggtacc tggtactgat tatggcactc aggaaagtat 2580
ctcgttactg gaagttagca ctctagggaa ggcaaaaaca gaaccaaata aatgtgtgag 2640
tcagtgtgca gcatttgaaa accccaaggg actaattcat ggttgttcca aagataatag 2700
aaatgacaca gaaggcttta agtatccatt gggacatgaa gttaaccaca gtcgggaaac 2760
aagcatagaa atggaagaaa gtgaacttga tgctcagtat ttgcagaata cattcaaggt 2820
ttcaaagcgc cagtcatttg ctccgttttc aaatccagga aatgcagaag aggaatgtgc 2880
aacattctct gcccactctg ggtccttaaa gaaacaaagt ccaaaagtca cttttgaatg 2940
tgaacaaaag gaagaaaatc aaggaaagaa tgagtctaat atcaagcctg tacagacagt 3000
taatatcact gcaggctttc ctgtggttgg tcagaaagat aagccagttg ataatgccaa 3060
atgtagtatc aaaggaggct ctaggttttg tctatcatct cagttcagag gcaacgaaac 3120
tggactcatt actccaaata aacatggact tttacaaaac ccatatcgta taccaccact 3180
ttttcccatc aagtcatttg ttaaaactaa atgtaagaaa aatctgctag aggaaaactt 3240
tgaggaacat tcaatgtcac ctgaaagaga aatgggaaat gagaacattc caagtacagt 3300
gagcacaatt agccgtaata acattagaga aaatgttttt aaagaagcca gctcaagcaa 3360
tattaatgaa gtaggttcca gtactaatga agtgggctcc agtattaatg aaataggttc 3420
cagtgatgaa aacattcaag cagaactagg tagaaacaga gggccaaaat tgaatgctat 3480
gcttagatta ggggttttgc aacctgaggt ctataaacaa agtcttcctg gaagtaattg 3540
taagcatcct gaaataaaaa agcaagaata tgaagaagta gttcagactg ttaatacaga 3600
tttctctcca tatctgattt cagataactt agaacagcct atgggaagta gtcatgcatc 3660
tcaggtttgt tctgagacac ctgatgacct gttagatgat ggtgaaataa aggaagatac 3720
tagttttgct gaaaatgaca ttaaggaaag ttctgctgtt tttagcaaaa gcgtccagaa 3780
aggagagctt agcaggagtc ctagcccttt cacccataca catttggctc agggttaccg 3840
aagaggggcc aagaaattag agtcctcaga agagaactta tctagtgagg atgaagagct 3900
tccctgcttc caacacttgt tatttggtaa agtaaacaat ataccttctc agtctactag 3960
gcatagcacc gttgctaccg agtgtctgtc taagaacaca gaggagaatt tattatcatt 4020
gaagaatagc ttaaatgact gcagtaacca ggtaatattg gcaaaggcat ctcaggaaca 4080
tcaccttagt gaggaaacaa aatgttctgc tagcttgttt tcttcacagt gcagtgaatt 4140
ggaagacttg actgcaaata caaacaccca ggatcctttc ttgattggtt cttccaaaca 4200
aatgaggcat cagtctgaaa gccagggagt tggtctgagt gacaaggaat tggtttcaga 4260
tgatgaagaa agaggaacgg gcttggaaga aaataatcaa gaagagcaaa gcatggattc 4320
aaacttaggt gaagcagcat ctgggtgtga gagtgaaaca agcgtctctg aagactgctc 4380
agggctatcc tctcagagtg acattttaac cactcagcag agggatacca tgcaacataa 4440
cctgataaag ctccagcagg aaatggctga actagaagct gtgttagaac agcatgggag 4500
ccagccttct aacagctacc cttccatcat aagtgactct tctgcccttg aggacctgcg 4560
aaatccagaa caaagcacat cagaaaaagc agtattaact tcacagaaaa gtagtgaata 4620
ccctataagc cagaatccag aaggcctttc tgctgacaag tttgaggtgt ctgcagatag 4680
ttctaccagt aaaaataaag aaccaggagt ggaaaggtca tccccttcta aatgcccatc 4740
attagatgat aggtggtaca tgcacagttg ctctgggagt cttcagaata gaaactaccc 4800
atctcaagag gagctcatta aggttgttga tgtggaggag caacagctgg aagagtctgg 4860
gccacacgat ttgacggaaa catcttactt gccaaggcaa gatctagagg gaacccctta 4920
cctggaatct ggaatcagcc tcttctctga tgaccctgaa tctgatcctt ctgaagacag 4980
agccccagag tcagctcgtg ttggcaacat accatcttca acctctgcat tgaaagttcc 5040
ccaattgaaa gttgcagaat ctgcccagag tccagctgct gctcatacta ctgatactgc 5100
tgggtataat gcaatggaag aaagtgtgag cagggagaag ccagaattga cagcttcaac 5160
agaaagggtc aacaaaagaa tgtccatggt ggtgtctggc ctgaccccag aagaatttat 5220
gctcgtgtac aagtttgcca gaaaacacca catcacttta actaatctaa ttactgaaga 5280
gactactcat gttgttatga aaacagatgc tgagtttgtg tgtgaacgga cactgaaata 5340
ttttctagga attgcgggag gaaaatgggt agttagctat ttctgggtga cccagtctat 5400
taaagaaaga aaaatgctga atgagcatga ttttgaagtc agaggagatg tggtcaatgg 5460
aagaaaccac caaggtccaa agcgagcaag agaatcccag gacagaaaga tcttcagggg 5520
gctagaaatc tgttgctatg ggcccttcac caacatgccc acagatcaac tggaatggat 5580
ggtacagctg tgtggtgctt ctgtggtgaa ggagctttca tcattcaccc ttggcacagg 5640
tgtccaccca attgtggttg tgcagccaga tgcctggaca gaggacaatg gcttccatgc 5700
aattgggcag atgtgtgagg cacctgtggt gacccgagag tgggtgttgg acagtgtagc 5760
actctaccag tgccaggagc tggacaccta cctgataccc cagatccccc acagccacta 5820
ctgactgcag ccagccacag gtacagagcc acaggacccc aagaatgagc ttacaaagtg 5880
gcctttccag gccctgggag ctcctctcac tcttcagtcc ttctactgtc ctggctacta 5940
aatattttat gtacatcagc ctgaaaagga cttctggcta tgcaagggtc ccttaaagat 6000
tttctgcttg aagtctccct tggaaatctg ccatgagcac aaaattatgg taatttttca 6060
cctgagaaga ttttaaaacc atttaaacgc caccaattga gcaagatgct gattcattat 6120
ttatcagccc tattctttct attcaggctg ttgttggctt agggctggaa gcacagagtg 6180
gcttggcctc aagagaatag ctggtttccc taagtttact tctctaaaac cctgtgttca 6240
caaaggcaga gagtcagacc cttcaatgga aggagagtgc ttgggatcga ttatgtgact 6300
taaagtcaga atagtccttg ggcagttctc aaatgttgga gtggaacatt ggggaggaaa 6360
ttctgaggca ggtattagaa atgaaaagga aacttgaaac ctgggcatgg tggctcacgc 6420
ctgtaatccc agcactttgg gaggccaagg tgggcagatc actggaggtc aggagttcga 6480
aaccagcctg gccaacatgg tgaaacccca tctctactaa aaatacagaa attagccggt 6540
catggtggtg gacacctgta atcccagcta ctcaggtggc taaggcagga gaatcacttc 6600
agcccgggag gtggaggttg cagtgagcca agatcatacc acggcactcc agcctgggtg 6660
acagtgagac tgtggctcaa aaaaaaaaaa aaaaaaagga aaatgaaact agaagagatt 6720
tctaaaagtc tgagatatat ttgctagatt tctaaagaat gtgttctaaa acagcagaag 6780
attttcaaga accggtttcc aaagacagtc ttctaattcc tcattagtaa taagtaaaat 6840
gtttattgtt gtagctctgg tatataatcc attcctctta aaatataaga cctctggcat 6900
gaatatttca tatctataaa atgacagatc ccaccaggaa ggaagctgtt gctttctttg 6960
aggtgatttt tttcctttgc tccctgttgc tgaaaccata cagcttcata aataattttg 7020
cttgctgaag gaagaaaaag tgtttttcat aaacccatta tccaggactg tttatagctg 7080
ttggaaggac taggtcttcc ctagcccccc cagtgtgcaa gggcagtgaa gacttgattg 7140
tacaaaatac gttttgtaaa tgttgtgctg ttaacactgc aaataaactt ggtagcaaac 7200
acttccaaaa aaaaaaaaaa aaaa 7224
<210> 13
<211> 24
<212> DNA
<213> 人工序列
<400> 13
aaaatcatct ctccgaaaaa caag 24
<210> 14
<211> 24
<212> DNA
<213> 人工序列
<400> 14
tgaaacacaa acgattttac cact 24
<210> 15
<211> 24
<212> DNA
<213> 人工序列
<400> 15
ataatgatga atgtagcacg catt 24
<210> 16
<211> 24
<212> DNA
<213> 人工序列
<400> 16
atactttgga aaagacttgc ttgg 24
<210> 17
<211> 25
<212> DNA
<213> 人工序列
<400> 17
gagaagcaaa atccactact aatgc 25
<210> 18
<211> 24
<212> DNA
<213> 人工序列
<400> 18
gaaaggtttg taccggtagt tgtt 24
<210> 19
<211> 11386
<212> DNA
<213> 人工序列
<400> 19
gtggcgcgag cttctgaaac taggcggcag aggcggagcc gctgtggcac tgctgcgcct 60
ctgctgcgcc tcgggtgtct tttgcggcgg tgggtcgccg ccgggagaag cgtgagggga 120
cagatttgtg accggcgcgg tttttgtcag cttactccgg ccaaaaaaga actgcacctc 180
tggagcggac ttatttacca agcattggag gaatatcgta ggtaaaaatg cctattggat 240
ccaaagagag gccaacattt tttgaaattt ttaagacacg ctgcaacaaa gcagatttag 300
gaccaataag tcttaattgg tttgaagaac tttcttcaga agctccaccc tataattctg 360
aacctgcaga agaatctgaa cataaaaaca acaattacga accaaaccta tttaaaactc 420
cacaaaggaa accatcttat aatcagctgg cttcaactcc aataatattc aaagagcaag 480
ggctgactct gccgctgtac caatctcctg taaaagaatt agataaattc aaattagact 540
taggaaggaa tgttcccaat agtagacata aaagtcttcg cacagtgaaa actaaaatgg 600
atcaagcaga tgatgtttcc tgtccacttc taaattcttg tcttagtgaa agtcctgttg 660
ttctacaatg tacacatgta acaccacaaa gagataagtc agtggtatgt gggagtttgt 720
ttcatacacc aaagtttgtg aagggtcgtc agacaccaaa acatatttct gaaagtctag 780
gagctgaggt ggatcctgat atgtcttggt caagttcttt agctacacca cccaccctta 840
gttctactgt gctcatagtc agaaatgaag aagcatctga aactgtattt cctcatgata 900
ctactgctaa tgtgaaaagc tatttttcca atcatgatga aagtctgaag aaaaatgata 960
gatttatcgc ttctgtgaca gacagtgaaa acacaaatca aagagaagct gcaagtcatg 1020
gatttggaaa aacatcaggg aattcattta aagtaaatag ctgcaaagac cacattggaa 1080
agtcaatgcc aaatgtccta gaagatgaag tatatgaaac agttgtagat acctctgaag 1140
aagatagttt ttcattatgt ttttctaaat gtagaacaaa aaatctacaa aaagtaagaa 1200
ctagcaagac taggaaaaaa attttccatg aagcaaacgc tgatgaatgt gaaaaatcta 1260
aaaaccaagt gaaagaaaaa tactcatttg tatctgaagt ggaaccaaat gatactgatc 1320
cattagattc aaatgtagca aatcagaagc cctttgagag tggaagtgac aaaatctcca 1380
aggaagttgt accgtctttg gcctgtgaat ggtctcaact aaccctttca ggtctaaatg 1440
gagcccagat ggagaaaata cccctattgc atatttcttc atgtgaccaa aatatttcag 1500
aaaaagacct attagacaca gagaacaaaa gaaagaaaga ttttcttact tcagagaatt 1560
ctttgccacg tatttctagc ctaccaaaat cagagaagcc attaaatgag gaaacagtgg 1620
taaataagag agatgaagag cagcatcttg aatctcatac agactgcatt cttgcagtaa 1680
agcaggcaat atctggaact tctccagtgg cttcttcatt tcagggtatc aaaaagtcta 1740
tattcagaat aagagaatca cctaaagaga ctttcaatgc aagtttttca ggtcatatga 1800
ctgatccaaa ctttaaaaaa gaaactgaag cctctgaaag tggactggaa atacatactg 1860
tttgctcaca gaaggaggac tccttatgtc caaatttaat tgataatgga agctggccag 1920
ccaccaccac acagaattct gtagctttga agaatgcagg tttaatatcc actttgaaaa 1980
agaaaacaaa taagtttatt tatgctatac atgatgaaac atcttataaa ggaaaaaaaa 2040
taccgaaaga ccaaaaatca gaactaatta actgttcagc ccagtttgaa gcaaatgctt 2100
ttgaagcacc acttacattt gcaaatgctg attcaggttt attgcattct tctgtgaaaa 2160
gaagctgttc acagaatgat tctgaagaac caactttgtc cttaactagc tcttttggga 2220
caattctgag gaaatgttct agaaatgaaa catgttctaa taatacagta atctctcagg 2280
atcttgatta taaagaagca aaatgtaata aggaaaaact acagttattt attaccccag 2340
aagctgattc tctgtcatgc ctgcaggaag gacagtgtga aaatgatcca aaaagcaaaa 2400
aagtttcaga tataaaagaa gaggtcttgg ctgcagcatg tcacccagta caacattcaa 2460
aagtggaata cagtgatact gactttcaat cccagaaaag tcttttatat gatcatgaaa 2520
atgccagcac tcttatttta actcctactt ccaaggatgt tctgtcaaac ctagtcatga 2580
tttctagagg caaagaatca tacaaaatgt cagacaagct caaaggtaac aattatgaat 2640
ctgatgttga attaaccaaa aatattccca tggaaaagaa tcaagatgta tgtgctttaa 2700
atgaaaatta taaaaacgtt gagctgttgc cacctgaaaa atacatgaga gtagcatcac 2760
cttcaagaaa ggtacaattc aaccaaaaca caaatctaag agtaatccaa aaaaatcaag 2820
aagaaactac ttcaatttca aaaataactg tcaatccaga ctctgaagaa cttttctcag 2880
acaatgagaa taattttgtc ttccaagtag ctaatgaaag gaataatctt gctttaggaa 2940
atactaagga acttcatgaa acagacttga cttgtgtaaa cgaacccatt ttcaagaact 3000
ctaccatggt tttatatgga gacacaggtg ataaacaagc aacccaagtg tcaattaaaa 3060
aagatttggt ttatgttctt gcagaggaga acaaaaatag tgtaaagcag catataaaaa 3120
tgactctagg tcaagattta aaatcggaca tctccttgaa tatagataaa ataccagaaa 3180
aaaataatga ttacatgaac aaatgggcag gactcttagg tccaatttca aatcacagtt 3240
ttggaggtag cttcagaaca gcttcaaata aggaaatcaa gctctctgaa cataacatta 3300
agaagagcaa aatgttcttc aaagatattg aagaacaata tcctactagt ttagcttgtg 3360
ttgaaattgt aaataccttg gcattagata atcaaaagaa actgagcaag cctcagtcaa 3420
ttaatactgt atctgcacat ttacagagta gtgtagttgt ttctgattgt aaaaatagtc 3480
atataacccc tcagatgtta ttttccaagc aggattttaa ttcaaaccat aatttaacac 3540
ctagccaaaa ggcagaaatt acagaacttt ctactatatt agaagaatca ggaagtcagt 3600
ttgaatttac tcagtttaga aaaccaagct acatattgca gaagagtaca tttgaagtgc 3660
ctgaaaacca gatgactatc ttaaagacca cttctgagga atgcagagat gctgatcttc 3720
atgtcataat gaatgcccca tcgattggtc aggtagacag cagcaagcaa tttgaaggta 3780
cagttgaaat taaacggaag tttgctggcc tgttgaaaaa tgactgtaac aaaagtgctt 3840
ctggttattt aacagatgaa aatgaagtgg ggtttagggg cttttattct gctcatggca 3900
caaaactgaa tgtttctact gaagctctgc aaaaagctgt gaaactgttt agtgatattg 3960
agaatattag tgaggaaact tctgcagagg tacatccaat aagtttatct tcaagtaaat 4020
gtcatgattc tgttgtttca atgtttaaga tagaaaatca taatgataaa actgtaagtg 4080
aaaaaaataa taaatgccaa ctgatattac aaaataatat tgaaatgact actggcactt 4140
ttgttgaaga aattactgaa aattacaaga gaaatactga aaatgaagat aacaaatata 4200
ctgctgccag tagaaattct cataacttag aatttgatgg cagtgattca agtaaaaatg 4260
atactgtttg tattcataaa gatgaaacgg acttgctatt tactgatcag cacaacatat 4320
gtcttaaatt atctggccag tttatgaagg agggaaacac tcagattaaa gaagatttgt 4380
cagatttaac ttttttggaa gttgcgaaag ctcaagaagc atgtcatggt aatacttcaa 4440
ataaagaaca gttaactgct actaaaacgg agcaaaatat aaaagatttt gagacttctg 4500
atacattttt tcagactgca agtgggaaaa atattagtgt cgccaaagag tcatttaata 4560
aaattgtaaa tttctttgat cagaaaccag aagaattgca taacttttcc ttaaattctg 4620
aattacattc tgacataaga aagaacaaaa tggacattct aagttatgag gaaacagaca 4680
tagttaaaca caaaatactg aaagaaagtg tcccagttgg tactggaaat caactagtga 4740
ccttccaggg acaacccgaa cgtgatgaaa agatcaaaga acctactcta ttgggttttc 4800
atacagctag cgggaaaaaa gttaaaattg caaaggaatc tttggacaaa gtgaaaaacc 4860
tttttgatga aaaagagcaa ggtactagtg aaatcaccag ttttagccat caatgggcaa 4920
agaccctaaa gtacagagag gcctgtaaag accttgaatt agcatgtgag accattgaga 4980
tcacagctgc cccaaagtgt aaagaaatgc agaattctct caataatgat aaaaaccttg 5040
tttctattga gactgtggtg ccacctaagc tcttaagtga taatttatgt agacaaactg 5100
aaaatctcaa aacatcaaaa agtatctttt tgaaagttaa agtacatgaa aatgtagaaa 5160
aagaaacagc aaaaagtcct gcaacttgtt acacaaatca gtccccttat tcagtcattg 5220
aaaattcagc cttagctttt tacacaagtt gtagtagaaa aacttctgtg agtcagactt 5280
cattacttga agcaaaaaaa tggcttagag aaggaatatt tgatggtcaa ccagaaagaa 5340
taaatactgc agattatgta ggaaattatt tgtatgaaaa taattcaaac agtactatag 5400
ctgaaaatga caaaaatcat ctctccgaaa aacaagatac ttatttaagt aacagtagca 5460
tgtctaacag ctattcctac cattctgatg aggtatataa tgattcagga tatctctcaa 5520
aaaataaact tgattctggt attgagccag tattgaagaa tgttgaagat caaaaaaaca 5580
ctagtttttc caaagtaata tccaatgtaa aagatgcaaa tgcataccca caaactgtaa 5640
atgaagatat ttgcgttgag gaacttgtga ctagctcttc accctgcaaa aataaaaatg 5700
cagccattaa attgtccata tctaatagta ataattttga ggtagggcca cctgcattta 5760
ggatagccag tggtaaaatc gtttgtgttt cacatgaaac aattaaaaaa gtgaaagaca 5820
tatttacaga cagtttcagt aaagtaatta aggaaaacaa cgagaataaa tcaaaaattt 5880
gccaaacgaa aattatggca ggttgttacg aggcattgga tgattcagag gatattcttc 5940
ataactctct agataatgat gaatgtagca cgcattcaca taaggttttt gctgacattc 6000
agagtgaaga aattttacaa cataaccaaa atatgtctgg attggagaaa gtttctaaaa 6060
tatcaccttg tgatgttagt ttggaaactt cagatatatg taaatgtagt atagggaagc 6120
ttcataagtc agtctcatct gcaaatactt gtgggatttt tagcacagca agtggaaaat 6180
ctgtccaggt atcagatgct tcattacaaa acgcaagaca agtgttttct gaaatagaag 6240
atagtaccaa gcaagtcttt tccaaagtat tgtttaaaag taacgaacat tcagaccagc 6300
tcacaagaga agaaaatact gctatacgta ctccagaaca tttaatatcc caaaaaggct 6360
tttcatataa tgtggtaaat tcatctgctt tctctggatt tagtacagca agtggaaagc 6420
aagtttccat tttagaaagt tccttacaca aagttaaggg agtgttagag gaatttgatt 6480
taatcagaac tgagcatagt cttcactatt cacctacgtc tagacaaaat gtatcaaaaa 6540
tacttcctcg tgttgataag agaaacccag agcactgtgt aaactcagaa atggaaaaaa 6600
cctgcagtaa agaatttaaa ttatcaaata acttaaatgt tgaaggtggt tcttcagaaa 6660
ataatcactc tattaaagtt tctccatatc tctctcaatt tcaacaagac aaacaacagt 6720
tggtattagg aaccaaagtg tcacttgttg agaacattca tgttttggga aaagaacagg 6780
cttcacctaa aaacgtaaaa atggaaattg gtaaaactga aactttttct gatgttcctg 6840
tgaaaacaaa tatagaagtt tgttctactt actccaaaga ttcagaaaac tactttgaaa 6900
cagaagcagt agaaattgct aaagctttta tggaagatga tgaactgaca gattctaaac 6960
tgccaagtca tgccacacat tctcttttta catgtcccga aaatgaggaa atggttttgt 7020
caaattcaag aattggaaaa agaagaggag agccccttat cttagtggga gaaccctcaa 7080
tcaaaagaaa cttattaaat gaatttgaca ggataataga aaatcaagaa aaatccttaa 7140
aggcttcaaa aagcactcca gatggcacaa taaaagatcg aagattgttt atgcatcatg 7200
tttctttaga gccgattacc tgtgtaccct ttcgcacaac taaggaacgt caagagatac 7260
agaatccaaa ttttaccgca cctggtcaag aatttctgtc taaatctcat ttgtatgaac 7320
atctgacttt ggaaaaatct tcaagcaatt tagcagtttc aggacatcca ttttatcaag 7380
tttctgctac aagaaatgaa aaaatgagac acttgattac tacaggcaga ccaaccaaag 7440
tctttgttcc accttttaaa actaaatcac attttcacag agttgaacag tgtgttagga 7500
atattaactt ggaggaaaac agacaaaagc aaaacattga tggacatggc tctgatgata 7560
gtaaaaataa gattaatgac aatgagattc atcagtttaa caaaaacaac tccaatcaag 7620
cagcagctgt aactttcaca aagtgtgaag aagaaccttt agatttaatt acaagtcttc 7680
agaatgccag agatatacag gatatgcgaa ttaagaagaa acaaaggcaa cgcgtctttc 7740
cacagccagg cagtctgtat cttgcaaaaa catccactct gcctcgaatc tctctgaaag 7800
cagcagtagg aggccaagtt ccctctgcgt gttctcataa acagctgtat acgtatggcg 7860
tttctaaaca ttgcataaaa attaacagca aaaatgcaga gtcttttcag tttcacactg 7920
aagattattt tggtaaggaa agtttatgga ctggaaaagg aatacagttg gctgatggtg 7980
gatggctcat accctccaat gatggaaagg ctggaaaaga agaattttat agggctctgt 8040
gtgacactcc aggtgtggat ccaaagctta tttctagaat ttgggtttat aatcactata 8100
gatggatcat atggaaactg gcagctatgg aatgtgcctt tcctaaggaa tttgctaata 8160
gatgcctaag cccagaaagg gtgcttcttc aactaaaata cagatatgat acggaaattg 8220
atagaagcag aagatcggct ataaaaaaga taatggaaag ggatgacaca gctgcaaaaa 8280
cacttgttct ctgtgtttct gacataattt cattgagcgc aaatatatct gaaacttcta 8340
gcaataaaac tagtagtgca gatacccaaa aagtggccat tattgaactt acagatgggt 8400
ggtatgctgt taaggcccag ttagatcctc ccctcttagc tgtcttaaag aatggcagac 8460
tgacagttgg tcagaagatt attcttcatg gagcagaact ggtgggctct cctgatgcct 8520
gtacacctct tgaagcccca gaatctctta tgttaaagat ttctgctaac agtactcggc 8580
ctgctcgctg gtataccaaa cttggattct ttcctgaccc tagacctttt cctctgccct 8640
tatcatcgct tttcagtgat ggaggaaatg ttggttgtgt tgatgtaatt attcaaagag 8700
cataccctat acagtggatg gagaagacat catctggatt atacatattt cgcaatgaaa 8760
gagaggaaga aaaggaagca gcaaaatatg tggaggccca acaaaagaga ctagaagcct 8820
tattcactaa aattcaggag gaatttgaag aacatgaaga aaacacaaca aaaccatatt 8880
taccatcacg tgcactaaca agacagcaag ttcgtgcttt gcaagatggt gcagagcttt 8940
atgaagcagt gaagaatgca gcagacccag cttaccttga gggttatttc agtgaagagc 9000
agttaagagc cttgaataat cacaggcaaa tgttgaatga taagaaacaa gctcagatcc 9060
agttggaaat taggaaggcc atggaatctg ctgaacaaaa ggaacaaggt ttatcaaggg 9120
atgtcacaac cgtgtggaag ttgcgtattg taagctattc aaaaaaagaa aaagattcag 9180
ttatactgag tatttggcgt ccatcatcag atttatattc tctgttaaca gaaggaaaga 9240
gatacagaat ttatcatctt gcaacttcaa aatctaaaag taaatctgaa agagctaaca 9300
tacagttagc agcgacaaaa aaaactcagt atcaacaact accggtttca gatgaaattt 9360
tatttcagat ttaccagcca cgggagcccc ttcacttcag caaattttta gatccagact 9420
ttcagccatc ttgttctgag gtggacctaa taggatttgt cgtttctgtt gtgaaaaaaa 9480
caggacttgc ccctttcgtc tatttgtcag acgaatgtta caatttactg gcaataaagt 9540
tttggataga ccttaatgag gacattatta agcctcatat gttaattgct gcaagcaacc 9600
tccagtggcg accagaatcc aaatcaggcc ttcttacttt atttgctgga gatttttctg 9660
tgttttctgc tagtccaaaa gagggccact ttcaagagac attcaacaaa atgaaaaata 9720
ctgttgagaa tattgacata ctttgcaatg aagcagaaaa caagcttatg catatactgc 9780
atgcaaatga tcccaagtgg tccaccccaa ctaaagactg tacttcaggg ccgtacactg 9840
ctcaaatcat tcctggtaca ggaaacaagc ttctgatgtc ttctcctaat tgtgagatat 9900
attatcaaag tcctttatca ctttgtatgg ccaaaaggaa gtctgtttcc acacctgtct 9960
cagcccagat gacttcaaag tcttgtaaag gggagaaaga gattgatgac caaaagaact 10020
gcaaaaagag aagagccttg gatttcttga gtagactgcc tttacctcca cctgttagtc 10080
ccatttgtac atttgtttct ccggctgcac agaaggcatt tcagccacca aggagttgtg 10140
gcaccaaata cgaaacaccc ataaagaaaa aagaactgaa ttctcctcag atgactccat 10200
ttaaaaaatt caatgaaatt tctcttttgg aaagtaattc aatagctgac gaagaacttg 10260
cattgataaa tacccaagct cttttgtctg gttcaacagg agaaaaacaa tttatatctg 10320
tcagtgaatc cactaggact gctcccacca gttcagaaga ttatctcaga ctgaaacgac 10380
gttgtactac atctctgatc aaagaacagg agagttccca ggccagtacg gaagaatgtg 10440
agaaaaataa gcaggacaca attacaacta aaaaatatat ctaagcattt gcaaaggcga 10500
caataaatta ttgacgctta acctttccag tttataagac tggaatataa tttcaaacca 10560
cacattagta cttatgttgc acaatgagaa aagaaattag tttcaaattt acctcagcgt 10620
ttgtgtatcg ggcaaaaatc gttttgcccg attccgtatt ggtatacttt tgcttcagtt 10680
gcatatctta aaactaaatg taatttatta actaatcaag aaaaacatct ttggctgagc 10740
tcggtggctc atgcctgtaa tcccaacact ttgagaagct gaggtgggag gagtgcttga 10800
ggccaggagt tcaagaccag cctgggcaac atagggagac ccccatcttt acaaagaaaa 10860
aaaaaagggg aaaagaaaat cttttaaatc tttggatttg atcactacaa gtattatttt 10920
acaagtgaaa taaacatacc attttctttt agattgtgtc attaaatgga atgaggtctc 10980
ttagtacagt tattttgatg cagataattc cttttagttt agctactatt ttaggggatt 11040
ttttttagag gtaactcact atgaaatagt tctccttaat gcaaatatgt tggttctgct 11100
atagttccat cctgttcaaa agtcaggatg aatatgaaga gtggtgtttc cttttgagca 11160
attcttcatc cttaagtcag catgattata agaaaaatag aaccctcagt gtaactctaa 11220
ttccttttta ctattccagt gtgatctctg aaattaaatt acttcaacta aaaattcaaa 11280
tactttaaat cagaagattt catagttaat ttattttttt tttcaacaaa atggtcatcc 11340
aaactcaaac ttgagaaaat atcttgcttt caaattggca ctgatt 11386
<210> 20
<211> 24
<212> DNA
<213> 人工序列
<400> 20
gaacactagg ctgcaaagac agta 24
<210> 21
<211> 24
<212> DNA
<213> 人工序列
<400> 21
caaagactct ccaagatggg atac 24
<210> 22
<211> 24
<212> DNA
<213> 人工序列
<400> 22
taacagtgac catatcaagc aggt 24
<210> 23
<211> 24
<212> DNA
<213> 人工序列
<400> 23
atgttctgtt ctccttcact ttcc 24
<210> 24
<211> 24
<212> DNA
<213> 人工序列
<400> 24
acgtcttcct tctctctctg tcat 24
<210> 25
<211> 24
<212> DNA
<213> 人工序列
<400> 25
tgtcaagaaa ctagtgctgg gtag 24
<210> 26
<211> 24
<212> DNA
<213> 人工序列
<400> 26
gaggaactgg atggagaaaa gtta 24
<210> 27
<211> 25
<212> DNA
<213> 人工序列
<400> 27
ttaaacaata cagctagtgg gaagg 25
<210> 28
<211> 5616
<212> DNA
<213> 人工序列
<400> 28
ccccggcgca gcgcggccgc agcagcctcc gccccccgca cggtgtgagc gcccgacgcg 60
gccgaggcgg ccggagtccc gagctagccc cggcggccgc cgccgcccag accggacgac 120
aggccacctc gtcggcgtcc gcccgagtcc ccgcctcgcc gccaacgcca caaccaccgc 180
gcacggcccc ctgactccgt ccagtattga tcgggagagc cggagcgagc tcttcgggga 240
gcagcgatgc gaccctccgg gacggccggg gcagcgctcc tggcgctgct ggctgcgctc 300
tgcccggcga gtcgggctct ggaggaaaag aaagtttgcc aaggcacgag taacaagctc 360
acgcagttgg gcacttttga agatcatttt ctcagcctcc agaggatgtt caataactgt 420
gaggtggtcc ttgggaattt ggaaattacc tatgtgcaga ggaattatga tctttccttc 480
ttaaagacca tccaggaggt ggctggttat gtcctcattg ccctcaacac agtggagcga 540
attcctttgg aaaacctgca gatcatcaga ggaaatatgt actacgaaaa ttcctatgcc 600
ttagcagtct tatctaacta tgatgcaaat aaaaccggac tgaaggagct gcccatgaga 660
aatttacagg aaatcctgca tggcgccgtg cggttcagca acaaccctgc cctgtgcaac 720
gtggagagca tccagtggcg ggacatagtc agcagtgact ttctcagcaa catgtcgatg 780
gacttccaga accacctggg cagctgccaa aagtgtgatc caagctgtcc caatgggagc 840
tgctggggtg caggagagga gaactgccag aaactgacca aaatcatctg tgcccagcag 900
tgctccgggc gctgccgtgg caagtccccc agtgactgct gccacaacca gtgtgctgca 960
ggctgcacag gcccccggga gagcgactgc ctggtctgcc gcaaattccg agacgaagcc 1020
acgtgcaagg acacctgccc cccactcatg ctctacaacc ccaccacgta ccagatggat 1080
gtgaaccccg agggcaaata cagctttggt gccacctgcg tgaagaagtg tccccgtaat 1140
tatgtggtga cagatcacgg ctcgtgcgtc cgagcctgtg gggccgacag ctatgagatg 1200
gaggaagacg gcgtccgcaa gtgtaagaag tgcgaagggc cttgccgcaa agtgtgtaac 1260
ggaataggta ttggtgaatt taaagactca ctctccataa atgctacgaa tattaaacac 1320
ttcaaaaact gcacctccat cagtggcgat ctccacatcc tgccggtggc atttaggggt 1380
gactccttca cacatactcc tcctctggat ccacaggaac tggatattct gaaaaccgta 1440
aaggaaatca cagggttttt gctgattcag gcttggcctg aaaacaggac ggacctccat 1500
gcctttgaga acctagaaat catacgcggc aggaccaagc aacatggtca gttttctctt 1560
gcagtcgtca gcctgaacat aacatccttg ggattacgct ccctcaagga gataagtgat 1620
ggagatgtga taatttcagg aaacaaaaat ttgtgctatg caaatacaat aaactggaaa 1680
aaactgtttg ggacctccgg tcagaaaacc aaaattataa gcaacagagg tgaaaacagc 1740
tgcaaggcca caggccaggt ctgccatgcc ttgtgctccc ccgagggctg ctggggcccg 1800
gagcccaggg actgcgtctc ttgccggaat gtcagccgag gcagggaatg cgtggacaag 1860
tgcaaccttc tggagggtga gccaagggag tttgtggaga actctgagtg catacagtgc 1920
cacccagagt gcctgcctca ggccatgaac atcacctgca caggacgggg accagacaac 1980
tgtatccagt gtgcccacta cattgacggc ccccactgcg tcaagacctg cccggcagga 2040
gtcatgggag aaaacaacac cctggtctgg aagtacgcag acgccggcca tgtgtgccac 2100
ctgtgccatc caaactgcac ctacggatgc actgggccag gtcttgaagg ctgtccaacg 2160
aatgggccta agatcccgtc catcgccact gggatggtgg gggccctcct cttgctgctg 2220
gtggtggccc tggggatcgg cctcttcatg cgaaggcgcc acatcgttcg gaagcgcacg 2280
ctgcggaggc tgctgcagga gagggagctt gtggagcctc ttacacccag tggagaagct 2340
cccaaccaag ctctcttgag gatcttgaag gaaactgaat tcaaaaagat caaagtgctg 2400
ggctccggtg cgttcggcac ggtgtataag ggactctgga tcccagaagg tgagaaagtt 2460
aaaattcccg tcgctatcaa ggaattaaga gaagcaacat ctccgaaagc caacaaggaa 2520
atcctcgatg aagcctacgt gatggccagc gtggacaacc cccacgtgtg ccgcctgctg 2580
ggcatctgcc tcacctccac cgtgcagctc atcacgcagc tcatgccctt cggctgcctc 2640
ctggactatg tccgggaaca caaagacaat attggctccc agtacctgct caactggtgt 2700
gtgcagatcg caaagggcat gaactacttg gaggaccgtc gcttggtgca ccgcgacctg 2760
gcagccagga acgtactggt gaaaacaccg cagcatgtca agatcacaga ttttgggctg 2820
gccaaactgc tgggtgcgga agagaaagaa taccatgcag aaggaggcaa agtgcctatc 2880
aagtggatgg cattggaatc aattttacac agaatctata cccaccagag tgatgtctgg 2940
agctacgggg tgaccgtttg ggagttgatg acctttggat ccaagccata tgacggaatc 3000
cctgccagcg agatctcctc catcctggag aaaggagaac gcctccctca gccacccata 3060
tgtaccatcg atgtctacat gatcatggtc aagtgctgga tgatagacgc agatagtcgc 3120
ccaaagttcc gtgagttgat catcgaattc tccaaaatgg cccgagaccc ccagcgctac 3180
cttgtcattc agggggatga aagaatgcat ttgccaagtc ctacagactc caacttctac 3240
cgtgccctga tggatgaaga agacatggac gacgtggtgg atgccgacga gtacctcatc 3300
ccacagcagg gcttcttcag cagcccctcc acgtcacgga ctcccctcct gagctctctg 3360
agtgcaacca gcaacaattc caccgtggct tgcattgata gaaatgggct gcaaagctgt 3420
cccatcaagg aagacagctt cttgcagcga tacagctcag accccacagg cgccttgact 3480
gaggacagca tagacgacac cttcctccca gtgcctgaat acataaacca gtccgttccc 3540
aaaaggcccg ctggctctgt gcagaatcct gtctatcaca atcagcctct gaaccccgcg 3600
cccagcagag acccacacta ccaggacccc cacagcactg cagtgggcaa ccccgagtat 3660
ctcaacactg tccagcccac ctgtgtcaac agcacattcg acagccctgc ccactgggcc 3720
cagaaaggca gccaccaaat tagcctggac aaccctgact accagcagga cttctttccc 3780
aaggaagcca agccaaatgg catctttaag ggctccacag ctgaaaatgc agaataccta 3840
agggtcgcgc cacaaagcag tgaatttatt ggagcatgac cacggaggat agtatgagcc 3900
ctaaaaatcc agactctttc gatacccagg accaagccac agcaggtcct ccatcccaac 3960
agccatgccc gcattagctc ttagacccac agactggttt tgcaacgttt acaccgacta 4020
gccaggaagt acttccacct cgggcacatt ttgggaagtt gcattccttt gtcttcaaac 4080
tgtgaagcat ttacagaaac gcatccagca agaatattgt ccctttgagc agaaatttat 4140
ctttcaaaga ggtatatttg aaaaaaaaaa aaagtatatg tgaggatttt tattgattgg 4200
ggatcttgga gtttttcatt gtcgctattg atttttactt caatgggctc ttccaacaag 4260
gaagaagctt gctggtagca cttgctaccc tgagttcatc caggcccaac tgtgagcaag 4320
gagcacaagc cacaagtctt ccagaggatg cttgattcca gtggttctgc ttcaaggctt 4380
ccactgcaaa acactaaaga tccaagaagg ccttcatggc cccagcaggc cggatcggta 4440
ctgtatcaag tcatggcagg tacagtagga taagccactc tgtcccttcc tgggcaaaga 4500
agaaacggag gggatggaat tcttccttag acttactttt gtaaaaatgt ccccacggta 4560
cttactcccc actgatggac cagtggtttc cagtcatgag cgttagactg acttgtttgt 4620
cttccattcc attgttttga aactcagtat gctgcccctg tcttgctgtc atgaaatcag 4680
caagagagga tgacacatca aataataact cggattccag cccacattgg attcatcagc 4740
atttggacca atagcccaca gctgagaatg tggaatacct aaggatagca ccgcttttgt 4800
tctcgcaaaa acgtatctcc taatttgagg ctcagatgaa atgcatcagg tcctttgggg 4860
catagatcag aagactacaa aaatgaagct gctctgaaat ctcctttagc catcacccca 4920
accccccaaa attagtttgt gttacttatg gaagatagtt ttctcctttt acttcacttc 4980
aaaagctttt tactcaaaga gtatatgttc cctccaggtc agctgccccc aaaccccctc 5040
cttacgcttt gtcacacaaa aagtgtctct gccttgagtc atctattcaa gcacttacag 5100
ctctggccac aacagggcat tttacaggtg cgaatgacag tagcattatg agtagtgtgg 5160
aattcaggta gtaaatatga aactagggtt tgaaattgat aatgctttca caacatttgc 5220
agatgtttta gaaggaaaaa agttccttcc taaaataatt tctctacaat tggaagattg 5280
gaagattcag ctagttagga gcccaccttt tttcctaatc tgtgtgtgcc ctgtaacctg 5340
actggttaac agcagtcctt tgtaaacagt gttttaaact ctcctagtca atatccaccc 5400
catccaattt atcaaggaag aaatggttca gaaaatattt tcagcctaca gttatgttca 5460
gtcacacaca catacaaaat gttccttttg cttttaaagt aatttttgac tcccagatca 5520
gtcagagccc ctacagcatt gttaagaaag tatttgattt ttgtctcaat gaaaataaaa 5580
ctatattcat ttccactcta aaaaaaaaaa aaaaaa 5616
<210> 29
<211> 24
<212> DNA
<213> 人工序列
<400> 29
tcacttttcc tttctaccca aaag 24
<210> 30
<211> 24
<212> DNA
<213> 人工序列
<400> 30
aagctgacaa cactagcaaa tgag 24
<210> 31
<211> 24
<212> DNA
<213> 人工序列
<400> 31
aaaatggttg ttgctgtgta aaaa 24
<210> 32
<211> 24
<212> DNA
<213> 人工序列
<400> 32
tgattcatca ggagagcatt taag 24
<210> 33
<211> 3927
<212> DNA
<213> 人工序列
<400> 33
taccgcgccg gagccttccg cagctgccgc ttcagtccga aggaggaagg gaaccaaccc 60
actttctcgg cgccgcggct cttttctaaa agtaatgtga aaacctttgc atcttctgat 120
agtctagcca aggtccaaga agtagcaagc tggcttttgg aaatgaatca ggaactgctc 180
tctgtgggca gcaaaagacg acgaactgga ggctctctga gaggtaaccc ttcctcaagc 240
caggtagatg aagaacagat gaatcgtgtg gtagaggagg aacagcaaca gcaactcaga 300
caacaagagg aggagcacac tgcaaggaat ggtgaagttg ttggagtaga acctagacct 360
ggaggccaaa atgattccca gcaaggacag ttggaagaaa acaataatag atttatttcg 420
gtagatgagg actcctcagg aaaccaagaa gaacaagagg aagatgaaga acatgctggt 480
gaacaagatg aggaggatga ggaggaggag gagatggacc aggagagtga cgattttgat 540
cagtctgatg atagtagcag agaagatgaa catacacata ctaacagtgt cacgaactcc 600
agtagtattg tggacctgcc cgttcaccaa ctctcctccc cattctatac aaaaacaaca 660
aaaatgaaaa gaaagttgga ccatggttct gaggtccgct ctttttcttt gggaaagaaa 720
ccatgcaaag tctcagaata tacaagtacc actgggcttg taccatgttc agcaacacca 780
acaacttttg gggacctcag agcagccaat ggccaagggc aacaacgacg ccgaattaca 840
tctgtccagc cacctacagg cctccaggaa tggctaaaaa tgtttcagag ctggagtgga 900
ccagagaaat tgcttgcttt agatgaactc attgatagtt gtgaaccaac acaagtaaaa 960
catatgatgc aagtgataga accccagttt caacgagact tcatttcatt gctccctaaa 1020
gagttggcac tctatgtgct ttcattcctg gaacccaaag acctgctaca agcagctcag 1080
acatgtcgct actggagaat tttggctgaa gacaaccttc tctggagaga gaaatgcaaa 1140
gaagagggga ttgatgaacc attgcacatc aagagaagaa aagtaataaa accaggtttc 1200
atacacagtc catggaaaag tgcatacatc agacagcaca gaattgatac taactggagg 1260
cgaggagaac tcaaatctcc taaggtgctg aaaggacatg atgatcatgt gatcacatgc 1320
ttacagtttt gtggtaaccg aatagttagt ggttctgatg acaacacttt aaaagtttgg 1380
tcagcagtca caggcaaatg tctgagaaca ttagtgggac atacaggtgg agtatggtca 1440
tcacaaatga gagacaacat catcattagt ggatctacag atcggacact caaagtgtgg 1500
aatgcagaga ctggagaatg tatacacacc ttatatgggc atacttccac tgtgcgttgt 1560
atgcatcttc atgaaaaaag agttgttagc ggttctcgag atgccactct tagggtttgg 1620
gatattgaga caggccagtg tttacatgtt ttgatgggtc atgttgcagc agtccgctgt 1680
gttcaatatg atggcaggag ggttgttagt ggagcatatg attttatggt aaaggtgtgg 1740
gatccagaga ctgaaacctg tctacacacg ttgcaggggc atactaatag agtctattca 1800
ttacagtttg atggtatcca tgtggtgagt ggatctcttg atacatcaat ccgtgtttgg 1860
gatgtggaga cagggaattg cattcacacg ttaacagggc accagtcgtt aacaagtgga 1920
atggaactca aagacaatat tcttgtctct gggaatgcag attctacagt taaaatctgg 1980
gatatcaaaa caggacagtg tttacaaaca ttgcaaggtc ccaacaagca tcagagtgct 2040
gtgacctgtt tacagttcaa caagaacttt gtaattacca gctcagatga tggaactgta 2100
aaactatggg acttgaaaac gggtgaattt attcgaaacc tagtcacatt ggagagtggg 2160
gggagtgggg gagttgtgtg gcggatcaga gcctcaaaca caaagctggt gtgtgcagtt 2220
gggagtcgga atgggactga agaaaccaag ctgctggtgc tggactttga tgtggacatg 2280
aagtgaagag cagaaaagat gaatttgtcc aattgtgtag acgatatact ccctgccctt 2340
ccccctgcaa aaagaaaaaa agaaaagaaa aagaaaaaaa tcccttgttc tcagtggtgc 2400
aggatgttgg cttggggcaa cagattgaaa agacctacag actaagaagg aaaagaagaa 2460
gagatgacaa accataactg acaagagagg cgtctgctgt ctcatcacat aaaaggcttc 2520
acttttgact gagggcagct ttgcaaaatg agactttcta aatcaaacca ggtgcaatta 2580
tttctttatt ttcttctcca gtggtcattg ggcagtgtta atgctgaaac atcattacag 2640
attctgctag cctgttcttt taccactgac agctagacac ctagaaagga actgcaataa 2700
tatcaaaaca agtactggtt gactttctaa ttagagagca tctgcaacaa aaagtcattt 2760
ttctggagtg gaaaagctta aaaaaattac tgtgaattgt ttttgtacag ttatcatgaa 2820
aagctttttt tttttttttt ttgccaacca ttgccaatgt caatcaatca cagtattagc 2880
ctctgttaat ctatttactg ttgcttccat atacattctt caatgcatat gttgctcaaa 2940
ggtggcaagt tgtcctgggt tctgtgagtc ctgagatgga tttaattctt gatgctggtg 3000
ctagaagtag gtcttcaaat atgggattgt tgtcccaacc ctgtactgta ctcccagtgg 3060
ccaaacttat ttatgctgct aaatgaaaga aagaaaaaag caaattattt ttttttattt 3120
tttttctgct gtgacgtttt agtcccagac tgaattccaa atttgctcta gtttggttat 3180
ggaaaaaaga ctttttgcca ctgaaacttg agccatctgt gcctctaaga ggctgagaat 3240
ggaagagttt cagataataa agagtgaagt ttgcctgcaa gtaaagaatt gagagtgtgt 3300
gcaaagctta ttttctttta tctgggcaaa aattaaaaca cattccttgg aacagagcta 3360
ttacttgcct gttctgtgga gaaacttttc tttttgaggg ctgtggtgaa tggatgaacg 3420
tacatcgtaa aactgacaaa atattttaaa aatatataaa acacaaaatt aaaataaagt 3480
tgctggtcag tcttagtgtt ttacagtatt tgggaaaaca actgttacag ttttattgct 3540
ctgagtaact gacaaagcag aaactattca gtttttgtag taaaggcgtc acatgcaaac 3600
aaacaaaatg aatgaaacag tcaaatggtt tgcctcattc tccaagagcc acaactcaag 3660
ctgaactgtg aaagtggttt aacactgtat cctaggcgat cttttttcct ccttctgttt 3720
atttttttgt ttgttttatt tatagtctga tttaaaacaa tcagattcaa gttggttaat 3780
tttagttatg taacaacctg acatgatgga ggaaaacaac ctttaaaggg attgtgtcta 3840
tggtttgatt cacttagaaa ttttattttc ttataactta agtgcaataa aatgtgtttt 3900
ttcatgttaa aaaaaaaaaa aaaaaaa 3927
<210> 34
<211> 19
<212> DNA
<213> 人工序列
<400> 34
gtgggtttgc ccttcagat 19
<210> 35
<211> 23
<212> DNA
<213> 人工序列
<400> 35
ctctagagga agcaggagac agg 23
<210> 36
<211> 21
<212> DNA
<213> 人工序列
<400> 36
ccctgtctcc tgcttcctct a 21
<210> 37
<211> 24
<212> DNA
<213> 人工序列
<400> 37
gatgtcctca aaagacttgg tgtt 24
<210> 38
<211> 1169
<212> DNA
<213> 人工序列
<400> 38
tgccctgcgc ccgcaacccg agccgcaccc gccgcggacg gagcccatgc gcggggcgaa 60
ccgcgcgccc ccgcccccgc cccgccccgg cctcggcccc ggccctggcc ccgggggcag 120
tcgcgcctgt gaacggtggg gcaggagacc ctgtaggagg accccgggcc gcaggcccct 180
gaggagcgat gacggaatat aagctggtgg tggtgggcgc cggcggtgtg ggcaagagtg 240
cgctgaccat ccagctgatc cagaaccatt ttgtggacga atacgacccc actatagagg 300
attcctaccg gaagcaggtg gtcattgatg gggagacgtg cctgttggac atcctggata 360
ccgccggcca ggaggagtac agcgccatgc gggaccagta catgcgcacc ggggagggct 420
tcctgtgtgt gtttgccatc aacaacacca agtcttttga ggacatccac cagtacaggg 480
agcagatcaa acgggtgaag gactcggatg acgtgcccat ggtgctggtg gggaacaagt 540
gtgacctggc tgcacgcact gtggaatctc ggcaggctca ggacctcgcc cgaagctacg 600
gcatccccta catcgagacc tcggccaaga cccggcaggg agtggaggat gccttctaca 660
cgttggtgcg tgagatccgg cagcacaagc tgcggaagct gaaccctcct gatgagagtg 720
gccccggctg catgagctgc aagtgtgtgc tctcctgacg caggtgaggg ggactcccag 780
ggcggccgcc acgcccaccg gatgaccccg gctccccgcc cctgccggtc tcctggcctg 840
cggtcagcag cctcccttgt gccccgccca gcacaagctc aggacatgga ggtgccggat 900
gcaggaagga ggtgcagacg gaaggaggag gaaggaagga cggaagcaag gaaggaagga 960
agggctgctg gagcccagtc accccgggac cgtgggccga ggtgactgca gaccctccca 1020
gggaggctgt gcacagactg tcttgaacat cccaaatgcc accggaaccc cagcccttag 1080
ctcccctccc aggcctctgt gggcccttgt cgggcacaga tgggatcaca gtaaattatt 1140
ggatggtctt gaaaaaaaaa aaaaaaaaa 1169
<210> 39
<211> 24
<212> DNA
<213> 人工序列
<400> 39
taaaacttgg cagattttgg tgta 24
<210> 40
<211> 24
<212> DNA
<213> 人工序列
<400> 40
aaatcacatt attgccaaca tgac 24
<210> 41
<211> 2402
<212> DNA
<213> 人工序列
<400> 41
gggctgagga ggcggggcct gggaggggac aaagccggga agaggaaaag ctcggaccta 60
ccctgtggtc ccgggtttct gcagagtcta cttcagaagc ggaggcactg ggagtccggt 120
ttgggattgc caggctgtgg ttgtgagtct gagcttgtga gcggctgtgg cgccccaact 180
cttcgccagc atatcatccc ggcaggcgat aaactacatt cagttgagtc tgcaagactg 240
ggaggaactg gggtgataag aaatctattc actgtcaagg tttattgaag tcaaaatgtc 300
caaaaaaatc agtggcggtt ctgtggtaga gatgcaagga gatgaaatga cacgaatcat 360
ttgggaattg attaaagaga aactcatttt tccctacgtg gaattggatc tacatagcta 420
tgatttaggc atagagaatc gtgatgccac caacgaccaa gtcaccaagg atgctgcaga 480
agctataaag aagcataatg ttggcgtcaa atgtgccact atcactcctg atgagaagag 540
ggttgaggag ttcaagttga aacaaatgtg gaaatcacca aatggcacca tacgaaatat 600
tctgggtggc acggtcttca gagaagccat tatctgcaaa aatatccccc ggcttgtgag 660
tggatgggta aaacctatca tcataggtcg tcatgcttat ggggatcaat acagagcaac 720
tgattttgtt gttcctgggc ctggaaaagt agagataacc tacacaccaa gtgacggaac 780
ccaaaaggtg acatacctgg tacataactt tgaagaaggt ggtggtgttg ccatggggat 840
gtataatcaa gataagtcaa ttgaagattt tgcacacagt tccttccaaa tggctctgtc 900
taagggttgg cctttgtatc tgagcaccaa aaacactatt ctgaagaaat atgatgggcg 960
ttttaaagac atctttcagg agatatatga caagcagtac aagtcccagt ttgaagctca 1020
aaagatctgg tatgagcata ggctcatcga cgacatggtg gcccaagcta tgaaatcaga 1080
gggaggcttc atctgggcct gtaaaaacta tgatggtgac gtgcagtcgg actctgtggc 1140
ccaagggtat ggctctctcg gcatgatgac cagcgtgctg gtttgtccag atggcaagac 1200
agtagaagca gaggctgccc acgggactgt aacccgtcac taccgcatgt accagaaagg 1260
acaggagacg tccaccaatc ccattgcttc catttttgcc tggaccagag ggttagccca 1320
cagagcaaag cttgataaca ataaagagct tgccttcttt gcaaatgctt tggaagaagt 1380
ctctattgag acaattgagg ctggcttcat gaccaaggac ttggctgctt gcattaaagg 1440
tttacccaat gtgcaacgtt ctgactactt gaatacattt gagttcatgg ataaacttgg 1500
agaaaacttg aagatcaaac tagctcaggc caaactttaa gttcatacct gagctaagaa 1560
ggataattgt cttttggtaa ctaggtctac aggtttacat ttttctgtgt tacactcaag 1620
gataaaggca aaatcaattt tgtaatttgt ttagaagcca gagtttatct tttctataag 1680
tttacagcct ttttcttata tatacagtta ttgccacctt tgtgaacatg gcaagggact 1740
tttttacaat ttttatttta ttttctagta ccagcctagg aattcggtta gtactcattt 1800
gtattcactg tcactttttc tcatgttcta attataaatg accaaaatca agattgctca 1860
aaagggtaaa tgatagccac agtattgctc cctaaaatat gcataaagta gaaattcact 1920
gccttcccct cctgtccatg accttgggca cagggaagtt ctggtgtcat agatatcccg 1980
ttttgtgagg tagagctgtg cattaaactt gcacatgact ggaacgaagt atgagtgcaa 2040
ctcaaatgtg ttgaagatac tgcagtcatt tttgtaaaga ccttgctgaa tgtttccaat 2100
agactaaata ctgtttaggc cgcaggagag tttggaatcc ggaataaata ctacctggag 2160
gtttgtcctc tccatttttc tctttctcct cctggcctgg cctgaatatt atactactct 2220
aaatagcata tttcatccaa gtgcaataat gtaagctgaa tcttttttgg acttctgctg 2280
gcctgtttta tttcttttat ataaatgtga tttctcagaa attgatatta aacactatct 2340
tatcttctcc tgaactgttg attttaatta aaattaagtg ctaattacca ttaaaaaaaa 2400
aa 2402
<210> 42
<211> 23
<212> DNA
<213> 人工序列
<400> 42
gtcgatggag gagtttgtaa atg 23
<210> 43
<211> 24
<212> DNA
<213> 人工序列
<400> 43
catgaaaatg gtcagagaaa cctt 24
<210> 44
<211> 24
<212> DNA
<213> 人工序列
<400> 44
tatgtggtag catctcatgg tttt 24
<210> 45
<211> 24
<212> DNA
<213> 人工序列
<400> 45
tatatgcatg gcattagcaa agac 24
<210> 46
<211> 5889
<212> DNA
<213> 人工序列
<400> 46
tcctaggcgg cggccgcggc ggcggaggca gcagcggcgg cggcagtggc ggcggcgaag 60
gtggcggcgg ctcggccagt actcccggcc cccgccattt cggactggga gcgagcgcgg 120
cgcaggcact gaaggcggcg gcggggccag aggctcagcg gctcccaggt gcgggagaga 180
ggcctgctga aaatgactga atataaactt gtggtagttg gagctggtgg cgtaggcaag 240
agtgccttga cgatacagct aattcagaat cattttgtgg acgaatatga tccaacaata 300
gaggattcct acaggaagca agtagtaatt gatggagaaa cctgtctctt ggatattctc 360
gacacagcag gtcaagagga gtacagtgca atgagggacc agtacatgag gactggggag 420
ggctttcttt gtgtatttgc cataaataat actaaatcat ttgaagatat tcaccattat 480
agagaacaaa ttaaaagagt taaggactct gaagatgtac ctatggtcct agtaggaaat 540
aaatgtgatt tgccttctag aacagtagac acaaaacagg ctcaggactt agcaagaagt 600
tatggaattc cttttattga aacatcagca aagacaagac agagagtgga ggatgctttt 660
tatacattgg tgagggagat ccgacaatac agattgaaaa aaatcagcaa agaagaaaag 720
actcctggct gtgtgaaaat taaaaaatgc attataatgt aatctgggtg ttgatgatgc 780
cttctataca ttagttcgag aaattcgaaa acataaagaa aagatgagca aagatggtaa 840
aaagaagaaa aagaagtcaa agacaaagtg tgtaattatg taaatacaat ttgtactttt 900
ttcttaaggc atactagtac aagtggtaat ttttgtacat tacactaaat tattagcatt 960
tgttttagca ttacctaatt tttttcctgc tccatgcaga ctgttagctt ttaccttaaa 1020
tgcttatttt aaaatgacag tggaagtttt tttttcctct aagtgccagt attcccagag 1080
ttttggtttt tgaactagca atgcctgtga aaaagaaact gaatacctaa gatttctgtc 1140
ttggggtttt tggtgcatgc agttgattac ttcttatttt tcttaccaat tgtgaatgtt 1200
ggtgtgaaac aaattaatga agcttttgaa tcatccctat tctgtgtttt atctagtcac 1260
ataaatggat taattactaa tttcagttga gaccttctaa ttggttttta ctgaaacatt 1320
gagggaacac aaatttatgg gcttcctgat gatgattctt ctaggcatca tgtcctatag 1380
tttgtcatcc ctgatgaatg taaagttaca ctgttcacaa aggttttgtc tcctttccac 1440
tgctattagt catggtcact ctccccaaaa tattatattt tttctataaa aagaaaaaaa 1500
tggaaaaaaa ttacaaggca atggaaacta ttataaggcc atttcctttt cacattagat 1560
aaattactat aaagactcct aatagctttt cctgttaagg cagacccagt atgaaatggg 1620
gattattata gcaaccattt tggggctata tttacatgct actaaatttt tataataatt 1680
gaaaagattt taacaagtat aaaaaattct cataggaatt aaatgtagtc tccctgtgtc 1740
agactgctct ttcatagtat aactttaaat cttttcttca acttgagtct ttgaagatag 1800
ttttaattct gcttgtgaca ttaaaagatt atttgggcca gttatagctt attaggtgtt 1860
gaagagacca aggttgcaag gccaggccct gtgtgaacct ttgagctttc atagagagtt 1920
tcacagcatg gactgtgtcc ccacggtcat ccagtgttgt catgcattgg ttagtcaaaa 1980
tggggaggga ctagggcagt ttggatagct caacaagata caatctcact ctgtggtggt 2040
cctgctgaca aatcaagagc attgcttttg tttcttaaga aaacaaactc ttttttaaaa 2100
attactttta aatattaact caaaagttga gattttgggg tggtggtgtg ccaagacatt 2160
aatttttttt ttaaacaatg aagtgaaaaa gttttacaat ctctaggttt ggctagttct 2220
cttaacactg gttaaattaa cattgcataa acacttttca agtctgatcc atatttaata 2280
atgctttaaa ataaaaataa aaacaatcct tttgataaat ttaaaatgtt acttatttta 2340
aaataaatga agtgagatgg catggtgagg tgaaagtatc actggactag gaagaaggtg 2400
acttaggttc tagataggtg tcttttagga ctctgatttt gaggacatca cttactatcc 2460
atttcttcat gttaaaagaa gtcatctcaa actcttagtt tttttttttt acaactatgt 2520
aatttatatt ccatttacat aaggatacac ttatttgtca agctcagcac aatctgtaaa 2580
tttttaacct atgttacacc atcttcagtg ccagtcttgg gcaaaattgt gcaagaggtg 2640
aagtttatat ttgaatatcc attctcgttt taggactctt cttccatatt agtgtcatct 2700
tgcctcccta ccttccacat gccccatgac ttgatgcagt tttaatactt gtaattcccc 2760
taaccataag atttactgct gctgtggata tctccatgaa gttttcccac tgagtcacat 2820
cagaaatgcc ctacatctta tttcctcagg gctcaagaga atctgacaga taccataaag 2880
ggatttgacc taatcactaa ttttcaggtg gtggctgatg ctttgaacat ctctttgctg 2940
cccaatccat tagcgacagt aggatttttc aaacctggta tgaatagaca gaaccctatc 3000
cagtggaagg agaatttaat aaagatagtg ctgaaagaat tccttaggta atctataact 3060
aggactactc ctggtaacag taatacattc cattgtttta gtaaccagaa atcttcatgc 3120
aatgaaaaat actttaattc atgaagctta cttttttttt ttggtgtcag agtctcgctc 3180
ttgtcaccca ggctggaatg cagtggcgcc atctcagctc actgcaacct ccatctccca 3240
ggttcaagcg attctcgtgc ctcggcctcc tgagtagctg ggattacagg cgtgtgccac 3300
tacactcaac taatttttgt atttttagga gagacggggt ttcaccctgt tggccaggct 3360
ggtctcgaac tcctgacctc aagtgattca cccaccttgg cctcataaac ctgttttgca 3420
gaactcattt attcagcaaa tatttattga gtgcctacca gatgccagtc accgcacaag 3480
gcactgggta tatggtatcc ccaaacaaga gacataatcc cggtccttag gtagtgctag 3540
tgtggtctgt aatatcttac taaggccttt ggtatacgac ccagagataa cacgatgcgt 3600
attttagttt tgcaaagaag gggtttggtc tctgtgccag ctctataatt gttttgctac 3660
gattccactg aaactcttcg atcaagctac tttatgtaaa tcacttcatt gttttaaagg 3720
aataaacttg attatattgt ttttttattt ggcataactg tgattctttt aggacaatta 3780
ctgtacacat taaggtgtat gtcagatatt catattgacc caaatgtgta atattccagt 3840
tttctctgca taagtaatta aaatatactt aaaaattaat agttttatct gggtacaaat 3900
aaacaggtgc ctgaactagt tcacagacaa ggaaacttct atgtaaaaat cactatgatt 3960
tctgaattgc tatgtgaaac tacagatctt tggaacactg tttaggtagg gtgttaagac 4020
ttacacagta cctcgtttct acacagagaa agaaatggcc atacttcagg aactgcagtg 4080
cttatgaggg gatatttagg cctcttgaat ttttgatgta gatgggcatt tttttaaggt 4140
agtggttaat tacctttatg tgaactttga atggtttaac aaaagatttg tttttgtaga 4200
gattttaaag ggggagaatt ctagaaataa atgttaccta attattacag ccttaaagac 4260
aaaaatcctt gttgaagttt ttttaaaaaa agctaaatta catagactta ggcattaaca 4320
tgtttgtgga agaatatagc agacgtatat tgtatcattt gagtgaatgt tcccaagtag 4380
gcattctagg ctctatttaa ctgagtcaca ctgcatagga atttagaacc taacttttat 4440
aggttatcaa aactgttgtc accattgcac aattttgtcc taatatatac atagaaactt 4500
tgtggggcat gttaagttac agtttgcaca agttcatctc atttgtattc cattgatttt 4560
ttttttcttc taaacatttt ttcttcaaac agtatataac tttttttagg ggattttttt 4620
ttagacagca aaaactatct gaagatttcc atttgtcaaa aagtaatgat ttcttgataa 4680
ttgtgtagta atgtttttta gaacccagca gttaccttaa agctgaattt atatttagta 4740
acttctgtgt taatactgga tagcatgaat tctgcattga gaaactgaat agctgtcata 4800
aaatgaaact ttctttctaa agaaagatac tcacatgagt tcttgaagaa tagtcataac 4860
tagattaaga tctgtgtttt agtttaatag tttgaagtgc ctgtttggga taatgatagg 4920
taatttagat gaatttaggg gaaaaaaaag ttatctgcag atatgttgag ggcccatctc 4980
tccccccaca cccccacaga gctaactggg ttacagtgtt ttatccgaaa gtttccaatt 5040
ccactgtctt gtgttttcat gttgaaaata cttttgcatt tttcctttga gtgccaattt 5100
cttactagta ctatttctta atgtaacatg tttacctgga atgtatttta actatttttg 5160
tatagtgtaa actgaaacat gcacattttg tacattgtgc tttcttttgt gggacatatg 5220
cagtgtgatc cagttgtttt ccatcatttg gttgcgctga cctaggaatg ttggtcatat 5280
caaacattaa aaatgaccac tcttttaatt gaaattaact tttaaatgtt tataggagta 5340
tgtgctgtga agtgatctaa aatttgtaat atttttgtca tgaactgtac tactcctaat 5400
tattgtaatg taataaaaat agttacagtg actatgagtg tgtatttatt catgaaattt 5460
gaactgtttg ccccgaaatg gatatggaat actttataag ccatagacac tatagtatac 5520
cagtgaatct tttatgcagc ttgttagaag tatcctttat ttctaaaagg tgctgtggat 5580
attatgtaaa ggcgtgtttg cttaaactta aaaccatatt tagaagtaga tgcaaaacaa 5640
atctgccttt atgacaaaaa aataggataa cattatttat ttatttcctt ttatcaaaga 5700
aggtaattga tacacaacag gtgacttggt tttaggccca aaggtagcag cagcaacatt 5760
aataatggaa ataattgaat agttagttat gtatgttaat gccagtcacc agcaggctat 5820
ttcaaggtca gaagtaatga ctccatacat attatttatt tctataacta catttaaatc 5880
attaccagg 5889
<210> 47
<211> 24
<212> DNA
<213> 人工序列
<400> 47
gggttgctag aaaactactc caga 24
<210> 48
<211> 24
<212> DNA
<213> 人工序列
<400> 48
taaagatgat ccgacaagtg agag 24
<210> 49
<211> 24
<212> DNA
<213> 人工序列
<400> 49
gaatttggtg ggttttcttt tatg 24
<210> 50
<211> 24
<212> DNA
<213> 人工序列
<400> 50
ttaatatccg caaatgactt gcta 24
<210> 51
<211> 4454
<212> DNA
<213> 人工序列
<400> 51
gaaacgtccc gtgtgggagg ggcgggtctg ggtgcggcct gccgcatgac tcgtggttcg 60
gaggcccacg tggccggggc ggggactcag gcgcctgggg cgccgactga ttacgtagcg 120
ggcggggccg gaagtgccgc tccttggtgg gggctgttca tggcggttcc ggggtctcca 180
acatttttcc cggctgtggt cctaaatctg tccaaagcag aggcagtgga gcttgaggtt 240
cttgctggtg tgaaatgact gagtacaaac tggtggtggt tggagcaggt ggtgttggga 300
aaagcgcact gacaatccag ctaatccaga accactttgt agatgaatat gatcccacca 360
tagaggattc ttacagaaaa caagtggtta tagatggtga aacctgtttg ttggacatac 420
tggatacagc tggacaagaa gagtacagtg ccatgagaga ccaatacatg aggacaggcg 480
aaggcttcct ctgtgtattt gccatcaata atagcaagtc atttgcggat attaacctct 540
acagggagca gattaagcga gtaaaagact cggatgatgt acctatggtg ctagtgggaa 600
acaagtgtga tttgccaaca aggacagttg atacaaaaca agcccacgaa ctggccaaga 660
gttacgggat tccattcatt gaaacctcag ccaagaccag acagggtgtt gaagatgctt 720
tttacacact ggtaagagaa atacgccagt accgaatgaa aaaactcaac agcagtgatg 780
atgggactca gggttgtatg ggattgccat gtgtggtgat gtaacaagat acttttaaag 840
ttttgtcaga aaagagccac tttcaagctg cactgacacc ctggtcctga cttccctgga 900
ggagaagtat tcctgttgct gtcttcagtc tcacagagaa gctcctgcta cttccccagc 960
tctcagtagt ttagtacaat aatctctatt tgagaagttc tcagaataac tacctcctca 1020
cttggctgtc tgaccagaga atgcacctct tgttactccc tgttattttt ctgccctggg 1080
ttcttccaca gcacaaacac acctctgcca ccccaggttt ttcatctgaa aagcagttca 1140
tgtctgaaac agagaaccaa accgcaaacg tgaaattcta ttgaaaacag tgtcttgagc 1200
tctaaagtag caactgctgg tgattttttt tttcttttta ctgttgaact tagaactatg 1260
ctaatttttg gagaaatgtc ataaattact gttttgccaa gaatatagtt attattgctg 1320
tttggtttgt ttataatgtt atcggctcta ttctctaaac tggcatctgc tctagattca 1380
taaatacaaa aatgaatact gaattttgag tctatcctag tcttcacaac tttgacgtaa 1440
ttaaatccaa ctttcacagt gaagtgcctt tttcctagaa gtggtttgta gacttccttt 1500
ataatatttc agtggaatag atgtctcaaa aatccttatg catgaaatga atgtctgaga 1560
tacgtctgtg acttatctac cattgaagga aagctatatc tatttgagag cagatgccat 1620
tttgtacatg tatgaaattg gttttccaga ggcctgtttt ggggctttcc caggagaaag 1680
atgaaactga aagcacatga ataatttcac ttaataattt ttacctaatc tccacttttt 1740
tcataggtta ctacctatac aatgtatgta atttgtttcc cctagcttac tgataaacct 1800
aatattcaat gaacttccat ttgtattcaa atttgtgtca taccagaaag ctctacattt 1860
gcagatgttc aaatattgta aaactttggt gcattgttat ttaatagctg tgatcagtga 1920
ttttcaaacc tcaaatatag tatattaaca aattacattt tcactgtata tcatggtatc 1980
ttaatgatgt atataattgc cttcaatccc cttctcaccc caccctctac agcttccccc 2040
acagcaatag gggcttgatt atttcagttg agtaaagcat ggtgctaatg gaccagggtc 2100
acagtttcaa aacttgaaca atccagttag catcacagag aaagaaattc ttctgcattt 2160
gctcattgca ccagtaactc cagctagtaa ttttgctagg tagctgcagt tagccctgca 2220
aggaaagaag aggtcagtta gcacaaaccc tttaccatga ctggaaaact cagtatcacg 2280
tatttaaaca tttttttttc ttttagccat gtagaaactc taaattaagc caatattctc 2340
atttgagaat gaggatgtct cagctgagaa acgttttaaa ttctctttat tcataatgtt 2400
ctttgaaggg tttaaaacaa gatgttgata aatctaagct gatgagtttg ctcaaaacag 2460
gaagttgaaa ttgttgagac aggaatggaa aatataatta attgatacct atgaggattt 2520
ggaggcttgg cattttaatt tgcagataat accctggtaa ttctcatgaa aaatagactt 2580
ggataacttt tgataaaaga ctaattccaa aatggccact ttgttcctgt ctttaatatc 2640
taaatactta ctgaggtcct ccatcttcta tattatgaat tttcatttat taagcaaatg 2700
tcatattacc ttgaaattca gaagagaaga aacatatact gtgtccagag tataatgaac 2760
ctgcagagtt gtgcttctta ctgctaattc tgggagcttt cacagtactg tcatcatttg 2820
taaatggaaa ttctgctttt ctgtttctgc tccttctgga gcagtgctac tctgtaattt 2880
tcctgaggct tatcacctca gtcatttctt ttttaaatgt ctgtgactgg cagtgattct 2940
ttttcttaaa aatctattaa atttgatgtc aaattaggga gaaagatagt tactcatctt 3000
gggctcttgt gccaatagcc cttgtatgta tgtacttaga gttttccaag tatgttctaa 3060
gcacagaagt ttctaaatgg ggccaaaatt cagacttgag tatgttcttt gaatacctta 3120
agaagttaca attagccggg catggtggcc cgtgcctgta gtcccagcta cttgagaggc 3180
tgaggcagga gaatcacttc aacccaggag gtggaggtta cagtgagcag agatcgtgcc 3240
actgcactcc agcctgggtg acaagagaga cttgtctcca aaaaaaaagt tacacctagg 3300
tgtgaatttt ggcacaaagg agtgacaaac ttatagttaa aagctgaata acttcagtgt 3360
ggtataaaac gtggttttta ggctatgttt gtgattgctg aaaagaattc tagtttacct 3420
caaaatcctt ctctttcccc aaattaagtg cctggccagc tgtcataaat tacatattcc 3480
ttttggtttt tttaaaggtt acatgttcaa gagtgaaaat aagatgttct gtctgaaggc 3540
taccatgccg gatctgtaaa tgaacctgtt aaatgctgta tttgctccaa cggcttacta 3600
tagaatgtta cttaatacaa tatcatactt attacaattt ttactatagg agtgtaatag 3660
gtaaaattaa tctctatttt agtgggccca tgtttagtct ttcaccatcc tttaaactgc 3720
tgtgaatttt tttgtcatga cttgaaagca aggatagaga aacactttag agatatgtgg 3780
ggttttttta ccattccaga gcttgtgagc ataatcatat ttgctttata tttatagtca 3840
tgaactccta agttggcagc tacaaccaag aaccaaaaaa tggtgcgttc tgcttcttgt 3900
aattcatctc tgctaataaa ttataagaag caaggaaaat tagggaaaat attttatttg 3960
gatggtttct ataaacaagg gactataatt cttgtacatt atttttcatc tttgctgttt 4020
ctttgagcag tctaatgtgc cacacaatta tctaaggtat ttgttttcta taagaattgt 4080
tttaaaagta ttcttgttac cagagtagtt gtattatatt tcaaaacgta agatgatttt 4140
taaaagcctg agtactgacc taagatggaa ttgtatgaac tctgctctgg agggagggga 4200
ggatgtccgt ggaagttgta agacttttat ttttttgtgc catcaaatat aggtaaaaat 4260
aattgtgcaa ttctgctgtt taaacaggaa ctattggcct ccttggccct aaatggaagg 4320
gccgatattt taagttgatt attttattgt aaattaatcc aacctagttc tttttaattt 4380
ggttgaatgt tttttcttgt taaatgatgt ttaaaaaata aaaactggaa gttcttggct 4440
tagtcataat tctt 4454
<210> 52
<211> 24
<212> DNA
<213> 人工序列
<400> 52
tggttctttc ctgtctctga aaat 24
<210> 53
<211> 24
<212> DNA
<213> 人工序列
<400> 53
tgagatcagc caaattcagt tatt 24
<210> 54
<211> 24
<212> DNA
<213> 人工序列
<400> 54
tcaggagatg tgttacaagg ctta 24
<210> 55
<211> 24
<212> DNA
<213> 人工序列
<400> 55
aattctaatg ctgttcatgg attg 24
<210> 56
<211> 3724
<212> DNA
<213> 人工序列
<400> 56
tctccctcgg cgccgccgcc gccgcccgcg gggctgggac ccgatgcggt tagagccgcg 60
gagcctggaa gagccccgag cgtttctgct ttgggacaac catacatcta attccttaaa 120
gtagttttat atgtaaaact tgcaaagaat cagaacaatg cctccacgac catcatcagg 180
tgaactgtgg ggcatccact tgatgccccc aagaatccta gtagaatgtt tactaccaaa 240
tggaatgata gtgactttag aatgcctccg tgaggctaca ttaataacca taaagcatga 300
actatttaaa gaagcaagaa aataccccct ccatcaactt cttcaagatg aatcttctta 360
cattttcgta agtgttactc aagaagcaga aagggaagaa ttttttgatg aaacaagacg 420
actttgtgac cttcggcttt ttcaaccctt tttaaaagta attgaaccag taggcaaccg 480
tgaagaaaag atcctcaatc gagaaattgg ttttgctatc ggcatgccag tgtgtgaatt 540
tgatatggtt aaagatccag aagtacagga cttccgaaga aatattctga acgtttgtaa 600
agaagctgtg gatcttaggg acctcaattc acctcatagt agagcaatgt atgtctatcc 660
tccaaatgta gaatcttcac cagaattgcc aaagcacata tataataaat tagataaagg 720
gcaaataata gtggtgatct gggtaatagt ttctccaaat aatgacaagc agaagtatac 780
tctgaaaatc aaccatgact gtgtaccaga acaagtaatt gctgaagcaa tcaggaaaaa 840
aactcgaagt atgttgctat cctctgaaca actaaaactc tgtgttttag aatatcaggg 900
caagtatatt ttaaaagtgt gtggatgtga tgaatacttc ctagaaaaat atcctctgag 960
tcagtataag tatataagaa gctgtataat gcttgggagg atgcccaatt tgatgttgat 1020
ggctaaagaa agcctttatt ctcaactgcc aatggactgt tttacaatgc catcttattc 1080
cagacgcatt tccacagcta caccatatat gaatggagaa acatctacaa aatccctttg 1140
ggttataaat agtgcactca gaataaaaat tctttgtgca acctacgtga atgtaaatat 1200
tcgagacatt gataagatct atgttcgaac aggtatctac catggaggag aacccttatg 1260
tgacaatgtg aacactcaaa gagtaccttg ttccaatccc aggtggaatg aatggctgaa 1320
ttatgatata tacattcctg atcttcctcg tgctgctcga ctttgccttt ccatttgctc 1380
tgttaaaggc cgaaagggtg ctaaagagga acactgtcca ttggcatggg gaaatataaa 1440
cttgtttgat tacacagaca ctctagtatc tggaaaaatg gctttgaatc tttggccagt 1500
acctcatgga ttagaagatt tgctgaaccc tattggtgtt actggatcaa atccaaataa 1560
agaaactcca tgcttagagt tggagtttga ctggttcagc agtgtggtaa agttcccaga 1620
tatgtcagtg attgaagagc atgccaattg gtctgtatcc cgagaagcag gatttagcta 1680
ttcccacgca ggactgagta acagactagc tagagacaat gaattaaggg aaaatgacaa 1740
agaacagctc aaagcaattt ctacacgaga tcctctctct gaaatcactg agcaggagaa 1800
agattttcta tggagtcaca gacactattg tgtaactatc cccgaaattc tacccaaatt 1860
gcttctgtct gttaaatgga attctagaga tgaagtagcc cagatgtatt gcttggtaaa 1920
agattggcct ccaatcaaac ctgaacaggc tatggaactt ctggactgta attacccaga 1980
tcctatggtt cgaggttttg ctgttcggtg cttggaaaaa tatttaacag atgacaaact 2040
ttctcagtat ttaattcagc tagtacaggt cctaaaatat gaacaatatt tggataactt 2100
gcttgtgaga tttttactga agaaagcatt gactaatcaa aggattgggc actttttctt 2160
ttggcattta aaatctgaga tgcacaataa aacagttagc cagaggtttg gcctgctttt 2220
ggagtcctat tgtcgtgcat gtgggatgta tttgaagcac ctgaataggc aagtcgaggc 2280
aatggaaaag ctcattaact taactgacat tctcaaacag gagaagaagg atgaaacaca 2340
aaaggtacag atgaagtttt tagttgagca aatgaggcga ccagatttca tggatgctct 2400
acagggcttt ctgtctcctc taaaccctgc tcatcaacta ggaaacctca ggcttgaaga 2460
gtgtcgaatt atgtcctctg caaaaaggcc actgtggttg aattgggaga acccagacat 2520
catgtcagag ttactgtttc agaacaatga gatcatcttt aaaaatgggg atgatttacg 2580
gcaagatatg ctaacacttc aaattattcg tattatggaa aatatctggc aaaatcaagg 2640
tcttgatctt cgaatgttac cttatggttg tctgtcaatc ggtgactgtg tgggacttat 2700
tgaggtggtg cgaaattctc acactattat gcaaattcag tgcaaaggcg gcttgaaagg 2760
tgcactgcag ttcaacagcc acacactaca tcagtggctc aaagacaaga acaaaggaga 2820
aatatatgat gcagccattg acctgtttac acgttcatgt gctggatact gtgtagctac 2880
cttcattttg ggaattggag atcgtcacaa tagtaacatc atggtgaaag acgatggaca 2940
actgtttcat atagattttg gacacttttt ggatcacaag aagaaaaaat ttggttataa 3000
acgagaacgt gtgccatttg ttttgacaca ggatttctta atagtgatta gtaaaggagc 3060
ccaagaatgc acaaagacaa gagaatttga gaggtttcag gagatgtgtt acaaggctta 3120
tctagctatt cgacagcatg ccaatctctt cataaatctt ttctcaatga tgcttggctc 3180
tggaatgcca gaactacaat cttttgatga cattgcatac attcgaaaga ccctagcctt 3240
agataaaact gagcaagagg ctttggagta tttcatgaaa caaatgaatg atgcacatca 3300
tggtggctgg acaacaaaaa tggattggat cttccacaca attaaacagc atgcattgaa 3360
ctgaaaagat aactgagaaa atgaaagctc actctggatt ccacactgca ctgttaataa 3420
ctctcagcag gcaaagaccg attgcatagg aattgcacaa tccatgaaca gcattagaat 3480
ttacagcaag aacagaaata aaatactata taatttaaat aatgtaaacg caaacagggt 3540
ttgatagcac ttaaactagt tcatttcaaa attaagcttt agaataatgc gcaatttcat 3600
gttatgcctt aagtccaaaa aggtaaactt tgaagattgt ttgtatcttt ttttaaaaaa 3660
caaaacaaaa caaaaatccc caaaatatat agaaatgatg gagaaggaaa aaaaaaaaaa 3720
aaaa 3724
<210> 57
<211> 24
<212> DNA
<213> 人工序列
<400> 57
ttaccacagt tgcacaatat cctt 24
<210> 58
<211> 25
<212> DNA
<213> 人工序列
<400> 58
cacctcaata aaactgaagg aaaaa 25
<210> 59
<211> 24
<212> DNA
<213> 人工序列
<400> 59
agtttgacag ttaaaggcat ttcc 24
<210> 60
<211> 24
<212> DNA
<213> 人工序列
<400> 60
cctctcatgt tacaatgcca taag 24
<210> 61
<211> 24
<212> DNA
<213> 人工序列
<400> 61
tacattcttc ataccaggac caga 24
<210> 62
<211> 24
<212> DNA
<213> 人工序列
<400> 62
caacgaagaa ttaggaaaag gaaa 24
<210> 63
<211> 5572
<212> DNA
<213> 人工序列
<400> 63
cctcccctcg cccggcgcgg tcccgtccgc ctctcgctcg cctcccgcct cccctcggtc 60
ttccgaggcg cccgggctcc cggcgcggcg gcggaggggg cgggcaggcc ggcgggcggt 120
gatgtggcgg gactctttat gcgctgcggc aggatacgcg ctcggcgctg ggacgcgact 180
gcgctcagtt ctctcctctc ggaagctgca gccatgatgg aagtttgaga gttgagccgc 240
tgtgaggcga ggccgggctc aggcgaggga gatgagagac ggcggcggcc gcggcccgga 300
gcccctctca gcgcctgtga gcagccgcgg gggcagcgcc ctcggggagc cggccggcct 360
gcggcggcgg cagcggcggc gtttctcgcc tcctcttcgt cttttctaac cgtgcagcct 420
cttcctcggc ttctcctgaa agggaaggtg gaagccgtgg gctcgggcgg gagccggctg 480
aggcgcggcg gcggcggcgg cacctcccgc tcctggagcg ggggggagaa gcggcggcgg 540
cggcggccgc ggcggctgca gctccaggga gggggtctga gtcgcctgtc accatttcca 600
gggctgggaa cgccggagag ttggtctctc cccttctact gcctccaaca cggcggcggc 660
ggcggcggca catccaggga cccgggccgg ttttaaacct cccgtccgcc gccgccgcac 720
cccccgtggc ccgggctccg gaggccgccg gcggaggcag ccgttcggag gattattcgt 780
cttctcccca ttccgctgcc gccgctgcca ggcctctggc tgctgaggag aagcaggccc 840
agtcgctgca accatccagc agccgccgca gcagccatta cccggctgcg gtccagagcc 900
aagcggcggc agagcgaggg gcatcagcta ccgccaagtc cagagccatt tccatcctgc 960
agaagaagcc ccgccaccag cagcttctgc catctctctc ctcctttttc ttcagccaca 1020
ggctcccaga catgacagcc atcatcaaag agatcgttag cagaaacaaa aggagatatc 1080
aagaggatgg attcgactta gacttgacct atatttatcc aaacattatt gctatgggat 1140
ttcctgcaga aagacttgaa ggcgtataca ggaacaatat tgatgatgta gtaaggtttt 1200
tggattcaaa gcataaaaac cattacaaga tatacaatct ttgtgctgaa agacattatg 1260
acaccgccaa atttaattgc agagttgcac aatatccttt tgaagaccat aacccaccac 1320
agctagaact tatcaaaccc ttttgtgaag atcttgacca atggctaagt gaagatgaca 1380
atcatgttgc agcaattcac tgtaaagctg gaaagggacg aactggtgta atgatatgtg 1440
catatttatt acatcggggc aaatttttaa aggcacaaga ggccctagat ttctatgggg 1500
aagtaaggac cagagacaaa aagggagtaa ctattcccag tcagaggcgc tatgtgtatt 1560
attatagcta cctgttaaag aatcatctgg attatagacc agtggcactg ttgtttcaca 1620
agatgatgtt tgaaactatt ccaatgttca gtggcggaac ttgcaatcct cagtttgtgg 1680
tctgccagct aaaggtgaag atatattcct ccaattcagg acccacacga cgggaagaca 1740
agttcatgta ctttgagttc cctcagccgt tacctgtgtg tggtgatatc aaagtagagt 1800
tcttccacaa acagaacaag atgctaaaaa aggacaaaat gtttcacttt tgggtaaata 1860
cattcttcat accaggacca gaggaaacct cagaaaaagt agaaaatgga agtctatgtg 1920
atcaagaaat cgatagcatt tgcagtatag agcgtgcaga taatgacaag gaatatctag 1980
tacttacttt aacaaaaaat gatcttgaca aagcaaataa agacaaagcc aaccgatact 2040
tttctccaaa ttttaaggtg aagctgtact tcacaaaaac agtagaggag ccgtcaaatc 2100
cagaggctag cagttcaact tctgtaacac cagatgttag tgacaatgaa cctgatcatt 2160
atagatattc tgacaccact gactctgatc cagagaatga accttttgat gaagatcagc 2220
atacacaaat tacaaaagtc tgaatttttt tttatcaaga gggataaaac accatgaaaa 2280
taaacttgaa taaactgaaa atggaccttt ttttttttaa tggcaatagg acattgtgtc 2340
agattaccag ttataggaac aattctcttt tcctgaccaa tcttgtttta ccctatacat 2400
ccacagggtt ttgacacttg ttgtccagtt gaaaaaaggt tgtgtagctg tgtcatgtat 2460
ataccttttt gtgtcaaaag gacatttaaa attcaattag gattaataaa gatggcactt 2520
tcccgtttta ttccagtttt ataaaaagtg gagacagact gatgtgtata cgtaggaatt 2580
ttttcctttt gtgttctgtc accaactgaa gtggctaaag agctttgtga tatactggtt 2640
cacatcctac ccctttgcac ttgtggcaac agataagttt gcagttggct aagagaggtt 2700
tccgaagggt tttgctacat tctaatgcat gtattcgggt taggggaatg gagggaatgc 2760
tcagaaagga aataatttta tgctggactc tggaccatat accatctcca gctatttaca 2820
cacacctttc tttagcatgc tacagttatt aatctggaca ttcgaggaat tggccgctgt 2880
cactgcttgt tgtttgcgca ttttttttta aagcatattg gtgctagaaa aggcagctaa 2940
aggaagtgaa tctgtattgg ggtacaggaa tgaaccttct gcaacatctt aagatccaca 3000
aatgaaggga tataaaaata atgtcatagg taagaaacac agcaacaatg acttaaccat 3060
ataaatgtgg aggctatcaa caaagaatgg gcttgaaaca ttataaaaat tgacaatgat 3120
ttattaaata tgttttctca attgtaacga cttctccatc tcctgtgtaa tcaaggccag 3180
tgctaaaatt cagatgctgt tagtacctac atcagtcaac aacttacact tattttacta 3240
gttttcaatc ataatacctg ctgtggatgc ttcatgtgct gcctgcaagc ttcttttttc 3300
tcattaaata taaaatattt tgtaatgctg cacagaaatt ttcaatttga gattctacag 3360
taagcgtttt ttttctttga agatttatga tgcacttatt caatagctgt cagccgttcc 3420
acccttttga ccttacacat tctattacaa tgaattttgc agttttgcac attttttaaa 3480
tgtcattaac tgttagggaa ttttacttga atactgaata catataatgt ttatattaaa 3540
aaggacattt gtgttaaaaa ggaaattaga gttgcagtaa actttcaatg ctgcacacaa 3600
aaaaaagaca tttgattttt cagtagaaat tgtcctacat gtgctttatt gatttgctat 3660
tgaaagaata gggttttttt tttttttttt tttttttttt ttaaatgtgc agtgttgaat 3720
catttcttca tagtgctccc ccgagttggg actagggctt caatttcact tcttaaaaaa 3780
aatcatcata tatttgatat gcccagactg catacgattt taagcggagt acaactacta 3840
ttgtaaagct aatgtgaaga tattattaaa aaggtttttt tttccagaaa tttggtgtct 3900
tcaaattata ccttcacctt gacatttgaa tatccagcca ttttgtttct taatggtata 3960
aaattccatt ttcaataact tattggtgct gaaattgttc actagctgtg gtctgaccta 4020
gttaatttac aaatacagat tgaataggac ctactagagc agcatttata gagtttgatg 4080
gcaaatagat taggcagaac ttcatctaaa atattcttag taaataatgt tgacacgttt 4140
tccatacctt gtcagtttca ttcaacaatt tttaaatttt taacaaagct cttaggattt 4200
acacatttat atttaaacat tgatatatag agtattgatt gattgctcat aagttaaatt 4260
ggtaaagtta gagacaacta ttctaacacc tcaccattga aatttatatg ccaccttgtc 4320
tttcataaaa gctgaaaatt gttacctaaa atgaaaatca acttcatgtt ttgaagatag 4380
ttataaatat tgttctttgt tacaatttcg ggcaccgcat attaaaacgt aactttattg 4440
ttccaatatg taacatggag ggccaggtca taaataatga cattataatg ggcttttgca 4500
ctgttattat ttttcctttg gaatgtgaag gtctgaatga gggttttgat tttgaatgtt 4560
tcaatgtttt tgagaagcct tgcttacatt ttatggtgta gtcattggaa atggaaaaat 4620
ggcattatat atattatata tataaatata tattatacat actctcctta ctttatttca 4680
gttaccatcc ccatagaatt tgacaagaat tgctatgact gaaaggtttt cgagtcctaa 4740
ttaaaacttt atttatggca gtattcataa ttagcctgaa atgcattctg taggtaatct 4800
ctgagtttct ggaatatttt cttagacttt ttggatgtgc agcagcttac atgtctgaag 4860
ttacttgaag gcatcacttt taagaaagct tacagttggg ccctgtacca tcccaagtcc 4920
tttgtagctc ctcttgaaca tgtttgccat acttttaaaa gggtagttga ataaatagca 4980
tcaccattct ttgctgtggc acaggttata aacttaagtg gagtttaccg gcagcatcaa 5040
atgtttcagc tttaaaaaat aaaagtaggg tacaagttta atgtttagtt ctagaaattt 5100
tgtgcaatat gttcataacg atggctgtgg ttgccacaaa gtgcctcgtt tacctttaaa 5160
tactgttaat gtgtcatgca tgcagatgga aggggtggaa ctgtgcacta aagtgggggc 5220
tttaactgta gtatttggca gagttgcctt ctacctgcca gttcaaaagt tcaacctgtt 5280
ttcatataga atatatatac taaaaaattt cagtctgtta aacagcctta ctctgattca 5340
gcctcttcag atactcttgt gctgtgcagc agtggctctg tgtgtaaatg ctatgcactg 5400
aggatacaca aaaataccaa tatgatgtgt acaggataat gcctcatccc aatcagatgt 5460
ccatttgtta ttgtgtttgt taacaaccct ttatctctta gtgttataaa ctccacttaa 5520
aactgattaa agtctcattc ttgtcaaaaa aaaaaaaaaa aaaaaaaaaa aa 5572
<210> 64
<211> 24
<212> DNA
<213> 人工序列
<400> 64
aaagaaaagc tcctgaggtg taga 24
<210> 65
<211> 24
<212> DNA
<213> 人工序列
<400> 65
taagagcaat cagtgaggaa tcag 24
<210> 66
<211> 24
<212> DNA
<213> 人工序列
<400> 66
tgattcctca ctgattgctc ttag 24
<210> 67
<211> 24
<212> DNA
<213> 人工序列
<400> 67
tatttttcag tagagacggg gttt 24
<210> 68
<211> 24
<212> DNA
<213> 人工序列
<400> 68
tgtgttatct cctaggttgg ctct 24
<210> 69
<211> 24
<212> DNA
<213> 人工序列
<400> 69
aaagccagag aaaagaaaac tgag 24
<210> 70
<211> 24
<212> DNA
<213> 人工序列
<400> 70
tgcttctctt ttcctatcct gagt 24
<210> 71
<211> 24
<212> DNA
<213> 人工序列
<400> 71
aaatattctc catccagtgg tttc 24
<210> 72
<211> 2591
<212> DNA
<213> 人工序列
<400> 72
gatgggattg gggttttccc ctcccatgtg ctcaagactg gcgctaaaag ttttgagctt 60
ctcaaaagtc tagagccacc gtccagggag caggtagctg ctgggctccg gggacacttt 120
gcgttcgggc tgggagcgtg ctttccacga cggtgacacg cttccctgga ttggcagcca 180
gactgccttc cgggtcactg ccatggagga gccgcagtca gatcctagcg tcgagccccc 240
tctgagtcag gaaacatttt cagacctatg gaaactactt cctgaaaaca acgttctgtc 300
ccccttgccg tcccaagcaa tggatgattt gatgctgtcc ccggacgata ttgaacaatg 360
gttcactgaa gacccaggtc cagatgaagc tcccagaatg ccagaggctg ctccccccgt 420
ggcccctgca ccagcagctc ctacaccggc ggcccctgca ccagccccct cctggcccct 480
gtcatcttct gtcccttccc agaaaaccta ccagggcagc tacggtttcc gtctgggctt 540
cttgcattct gggacagcca agtctgtgac ttgcacgtac tcccctgccc tcaacaagat 600
gttttgccaa ctggccaaga cctgccctgt gcagctgtgg gttgattcca cacccccgcc 660
cggcacccgc gtccgcgcca tggccatcta caagcagtca cagcacatga cggaggttgt 720
gaggcgctgc ccccaccatg agcgctgctc agatagcgat ggtctggccc ctcctcagca 780
tcttatccga gtggaaggaa atttgcgtgt ggagtatttg gatgacagaa acacttttcg 840
acatagtgtg gtggtgccct atgagccgcc tgaggttggc tctgactgta ccaccatcca 900
ctacaactac atgtgtaaca gttcctgcat gggcggcatg aaccggaggc ccatcctcac 960
catcatcaca ctggaagact ccagtggtaa tctactggga cggaacagct ttgaggtgcg 1020
tgtttgtgcc tgtcctggga gagaccggcg cacagaggaa gagaatctcc gcaagaaagg 1080
ggagcctcac cacgagctgc ccccagggag cactaagcga gcactgccca acaacaccag 1140
ctcctctccc cagccaaaga agaaaccact ggatggagaa tatttcaccc ttcagatccg 1200
tgggcgtgag cgcttcgaga tgttccgaga gctgaatgag gccttggaac tcaaggatgc 1260
ccaggctggg aaggagccag gggggagcag ggctcactcc agccacctga agtccaaaaa 1320
gggtcagtct acctcccgcc ataaaaaact catgttcaag acagaagggc ctgactcaga 1380
ctgacattct ccacttcttg ttccccactg acagcctccc acccccatct ctccctcccc 1440
tgccattttg ggttttgggt ctttgaaccc ttgcttgcaa taggtgtgcg tcagaagcac 1500
ccaggacttc catttgcttt gtcccggggc tccactgaac aagttggcct gcactggtgt 1560
tttgttgtgg ggaggaggat ggggagtagg acataccagc ttagatttta aggtttttac 1620
tgtgagggat gtttgggaga tgtaagaaat gttcttgcag ttaagggtta gtttacaatc 1680
agccacattc taggtagggg cccacttcac cgtactaacc agggaagctg tccctcactg 1740
ttgaattttc tctaacttca aggcccatat ctgtgaaatg ctggcatttg cacctacctc 1800
acagagtgca ttgtgagggt taatgaaata atgtacatct ggccttgaaa ccacctttta 1860
ttacatgggg tctagaactt gacccccttg agggtgcttg ttccctctcc ctgttggtcg 1920
gtgggttggt agtttctaca gttgggcagc tggttaggta gagggagttg tcaagtctct 1980
gctggcccag ccaaaccctg tctgacaacc tcttggtgaa ccttagtacc taaaaggaaa 2040
tctcacccca tcccacaccc tggaggattt catctcttgt atatgatgat ctggatccac 2100
caagacttgt tttatgctca gggtcaattt cttttttctt tttttttttt ttttttcttt 2160
ttctttgaga ctgggtctcg ctttgttgcc caggctggag tggagtggcg tgatcttggc 2220
ttactgcagc ctttgcctcc ccggctcgag cagtcctgcc tcagcctccg gagtagctgg 2280
gaccacaggt tcatgccacc atggccagcc aacttttgca tgttttgtag agatggggtc 2340
tcacagtgtt gcccaggctg gtctcaaact cctgggctca ggcgatccac ctgtctcagc 2400
ctcccagagt gctgggatta caattgtgag ccaccacgtc cagctggaag ggtcaacatc 2460
ttttacattc tgcaagcaca tctgcatttt caccccaccc ttcccctcct tctccctttt 2520
tatatcccat ttttatatcg atctcttatt ttacaataaa actttgctgc cacctgtgtg 2580
tctgaggggt g 2591
Claims (8)
1.人肿瘤预测性生物标志组合物，其特征在于包括BRAF、BRCA1、BRCA2、EGFR、FBXW7、H-RAS、IDH1、K-RAS、N-RAS、PIK3CA、PTEN和TP53基因中的至少两个。
2.根据权利要求1所述人肿瘤预测性生物标志组合物，其特征在于组合物中含有2、4、6或12个所述基因。
3.权利要求1或2所述人肿瘤预测性生物标志组合物在制备肿瘤预测或筛查试剂盒中的应用。
4.根据权利要求3所述的应用，其特征在于所述试剂盒中包括组合物所含基因的对应引物。
5.一种人肿瘤诊断试剂盒，其特征在于含有检测BRAF、BRCA1、BRCA2、EGFR、FBXW7、H-RAS、IDH1、K-RAS、N-RAS、PIK3CA、PTEN和TP53中至少两个基因的引物。
6.根据权利要求5所述的人肿瘤诊断试剂盒，其特征在于含有2、4、6或12个所述基因的引物。
7.根据权利要求5或6所述的人肿瘤诊断试剂盒，其特征在于检测BRAF的引物序列如SEQ ID NO.1~2所示；检测BRCA1的引物序列如SEQ ID NO.4~11所示；检测BRCA2的引物序列如SEQ ID NO.13~18所示；检测EGFR的引物序列如SEQ ID NO.20~27所示；检测FBXW7的引物序列如SEQ ID NO.29~32所示；检测H-RAS的引物序列如SEQ ID NO.34~37所示；检测IDH1的引物序列为如SEQ ID NO.39~40所示；检测K-RAS的引物序列如SEQ ID NO.42~45所示；检测N-RAS的引物序列如SEQ ID NO.47~50所示；检测PIK3CA的引物序列如SEQ ID NO.52~55所示；检测PTEN的引物序列如SEQ ID NO.57~62所示；检测TP53的引物序列如SEQ ID NO.64~71所示。
8.根据权利要求7所述的人肿瘤诊断试剂盒，其特征在于所述试剂盒中还含有其它必要试剂：dNTPs、MgCl2、反应缓冲液和Taq DNA聚合酶。
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| CN105112515A (zh) * | 2015-08-14 | 2015-12-02 | 深圳市众偱精准医学研究院 | Fbxw7基因及表达产物在肾癌检测、肾癌药物制备中的用途 |
| CN105238799A (zh) * | 2015-10-19 | 2016-01-13 | 昆明理工大学 | 一种乳腺癌pik3ca突变基因及其应用 |
| CN107090503A (zh) * | 2017-04-27 | 2017-08-25 | 元码基因科技（北京）有限公司 | 探针组合物、基因捕获芯片、试剂盒及其应用 |
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| CN105112515A (zh) * | 2015-08-14 | 2015-12-02 | 深圳市众偱精准医学研究院 | Fbxw7基因及表达产物在肾癌检测、肾癌药物制备中的用途 |
| CN105238799A (zh) * | 2015-10-19 | 2016-01-13 | 昆明理工大学 | 一种乳腺癌pik3ca突变基因及其应用 |
| CN105238799B (zh) * | 2015-10-19 | 2018-08-31 | 昆明理工大学 | 一种乳腺癌pik3ca突变基因及其应用 |
| CN107419001A (zh) * | 2016-05-23 | 2017-12-01 | 益善生物技术股份有限公司 | Brca1、pten基因异常检测试剂盒及检测方法 |
| CN107090503A (zh) * | 2017-04-27 | 2017-08-25 | 元码基因科技（北京）有限公司 | 探针组合物、基因捕获芯片、试剂盒及其应用 |
| CN107090503B (zh) * | 2017-04-27 | 2020-06-19 | 元码基因科技（北京）股份有限公司 | 探针组合物、基因捕获芯片、试剂盒及其应用 |
| CN110760575A (zh) * | 2018-07-27 | 2020-02-07 | 中国人民解放军军事科学院军事医学研究院 | 一种用于检测微波辐射致脑损伤的生物标志物Braf蛋白 |
| CN112708665A (zh) * | 2019-10-25 | 2021-04-27 | 益善生物技术股份有限公司 | 用于肺癌多基因突变测序文库的构建方法与试剂盒 |
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