CN113943784A - 一种特异性识别iyvlvmlvl肽段的荧光pcr引物组及其应用 - Google Patents
一种特异性识别iyvlvmlvl肽段的荧光pcr引物组及其应用 Download PDFInfo
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Abstract
本发明属于基因工程技术领域,尤其涉及一种特异性识别IYVLVMLVL肽段的荧光PCR引物组及其应用。本申请的引物组是根据HLAA24分型,EBV病毒抗原肽段IYVLVMLVL的TCR设计的包括位点为L24‑49和/或L24‑2引物对。本申请的引物组能特异性的识别HLAA24分型患者体内EBV病毒抗原肽段IYVLVMLVL的TCR,为追踪监测患者体内TCR‑T细胞药物提供依据。
Description
本发明要求申请号为202110687266.8,申请日为2021年06月21日,发明名称为“一种特异性识别IYVLVMLVL肽段的荧光PCR引物组及其应用”的中国专利的优选权。
技术领域
本发明涉及基因工程技术领域,尤其涉及一种特异性识别IYVLVMLVL肽段的荧光PCR引物组及其应用。
背景技术
EBV是可以感染人类的疱疹病毒家族的一员。EBV感染与某些类型的癌症有关。这些因EBV病毒感染所导致的癌症患者有望在TCR-T细胞治疗中获益。
TCR-T细胞治疗是有望治疗实体肿瘤的新一代免疫细胞疗法。在TCR-T细胞治疗的过程中,需要不断监测被输注到病人体内的药用TCR-T细胞的生长和续存的情况。能实现此目的技术手段中,荧光定量PCR(qPCR)是检测灵敏度很高的技术手段。利用qPCR监测TCR-T细胞,需要针对TCR-T细胞的特定TCR基因序列设计qPCR引物。然而现有技术并未发现针对TCR基因序列所设计的qPCR引物组应用于检测TCR-T细胞在病人体内生长和续存状况中。
发明内容
针对现有技术的缺陷,本发明的目的在于提供一种特异灵敏的追踪TCR-T细胞药物的引物组,尤其涉及一种特异性识别IYVLVMLVL肽段的荧光PCR引物组及其应用。
为了实现上述发明目的,本发明提供以下技术方案:
本发明提供了一种TCR引物组,包括位点为L24-2和/或L24-49的引物对。
作为优选,所述引物组依据HLAA24分型,EBV病毒抗原肽段IYVLVMLVL的TCR设计。
作为优选,位点为L24-2的TCR引物对的序列如SEQ ID NO.1~2所示;
位点为L24-49的TCR引物对的序列如SEQ ID NO.3~4所示。
作为优选,位点为L24-2的TCR引物对的序列还如SEQ ID NO.6~7所示;
位点为L24-49的TCR引物对的序列还如SEQ ID NO.8~9所示。
本发明还提供了所述的引物组在制备特异性检测HLAA24分型患者体内EBV病毒抗原肽段IYVLVMLVL的TCR产品中的应用。
作为优选,所述产品为试剂盒产品。
作为优选,所述试剂盒为荧光定量PCR试剂盒。
本发明提供了一种特异性识别IYVLVMLVL肽段的荧光PCR引物组及其应用。本申请的引物组是根据HLAA24分型,EBV病毒抗原肽段IYVLVMLVL的TCR设计的。将本申请的引物组制备成荧光定量PCR试剂盒,追踪患者体内TCR-T细胞药物时,能特异的检测到药物的灵敏度为27个拷贝数的基因片段。
附图说明
图1为L24-2位点质粒DNA标准品的扩增曲线(从左到右质粒DNA标准品的稀释度分别为108拷贝、107拷贝、106拷贝、105拷贝、104拷贝、103拷贝、102拷贝、101拷贝的基因片段)。
图2为L24-2位点质粒DNA标准品扩增产物的电泳图(从左到右质粒DNA标准品的稀释度分别108拷贝、107拷贝、106拷贝、105拷贝、104拷贝、103拷贝、102拷贝、101拷贝的基因片段)。
图3为L24-2位点质粒DNA标准品的标准曲线图。
图4为L24-2位点的TCR-T细胞药的基因组DNA的扩增曲线(从左到右,基因组DNA的稀释度分别为原液、10倍稀释、100倍稀释、1000倍稀释、10000倍稀释)。
图5为L24-2位点的TCR-T细胞药的基因组DNA扩增产物的电泳图(从左到右基因组DNA的稀释度分别为原液、10倍稀释、100倍稀释、1000倍稀释、10000倍稀释)。
具体实施方式
在本发明中,所述位点为L24-2的TCR引物对的模板序列如SEQ ID NO.10所示;
所述位点为L24-49的TCR引物对的模板序列如SEQ ID NO.11所示。
本发明实施例中所述的TCR质粒标准品的构建方法为:
(1)将每个位点对应的TCR基因片段克隆至SEQ ID NO.5所示的质粒载体中,得到含有特定TCR基因片段的质粒载体;
(2)将含有特定TCR基因片段的质粒载体在大肠杆菌内扩增,提纯质粒DNA,得到相应位点的TCR质粒标准品。
在本发明实施例中以L24-2位点为例设计方案,其余位点的检测方法与之相同。
在本发明实施例中所述的TCR-T细胞药的基因组DNA为从相应的TCR-T药物细胞中提取的基因组DNA。
下面结合实施例对本发明提供的技术方案进行详细的说明,但是不能把它们理解为对本发明保护范围的限定。
实施例1引物设计
根据HLAA24分型,EBV病毒抗原肽段IYVLVMLVL的L24-2和L24-49位点设计得到的TCR引物。得到的引物的序列如SEQ ID NO.1~4和SEQ ID NO.6~9所示。
实施例2标准曲线建立
以TCR的L24-2位点为例设计方案。
取L24-2位点的质粒DNA标准品的108个拷贝数稀释液、107个拷贝数稀释液、106个拷贝数稀释液、105个拷贝数稀释液、104个拷贝数稀释液、103个拷贝数稀释液、102个拷贝数稀释液、101个拷贝数稀释液,按照下述的反应体系和反应程序设置实时荧光PCR。
反应体系:PCR预混液10μl;正向引物0.4μl,引物浓度:10μm/μl;反向引物0.4μl,引物浓度:10μm/μl;ROX Reference Dye(50×)0.2μl;TCR-T细胞药的基因组DNA2μl,浓度50ng/μl,或TCR的质粒标准品DNA2μl;Nuclease-Free Water 7μl;总反应体系共20μl。
PCR反应程序:95℃,30s;(95℃,5s;60℃,20s)40个循环。设置参比荧光为ROX,选择反应类型为SYBR Green Reagents,选择荧光检测通道SYBR,设置在60℃时采集荧光信号。
经实时荧光PCR扩增得到的L24-2位点质粒DNA标准品的扩增曲线如图1所示。
将每个稀释度的质粒DNA标准品的扩增产物进行琼脂糖凝胶电泳,结果如图2所示。
以扩增曲线的Ct值为纵坐标,质粒DNA标准品的log以10为底的基因片段拷贝数为横坐标建立标准曲线,标准曲线的结果如图3所示。计算公式为:y=-3.5555x+41.485(R2=0.9981)
由于计算公式中的R2≥0.95,则证明本实施例的标准曲线有效。
实施例3灵敏度检测
将针对L24-2位点的TCR-T细胞药的基因组DNA原液、10倍稀释液、100倍稀释液、1000倍稀释液、10000倍稀释液,根据实施例2实时荧光PCR的反应体系和程序进行扩增。得到针对L24-2位点的TCR-T细胞药的基因组DNA的扩增曲线如图4所示。将扩增产物进行琼脂糖凝胶电泳,结果如图5所示。
将得到的每个稀释度的TCR-T细胞药的基因组DNA的Ct值代到实施例2的标准曲线中,可以计算得到每个稀释度的TCR-T细胞药的基因组DNA的拷贝数。
经过计算,针对L24-2位点的TCR-T细胞药基因组DNA原液的拷贝数为46766个拷贝,10倍稀释液的拷贝数为7998个拷贝、100倍稀释液的拷贝数为873个拷贝、1000倍稀释液的拷贝数为125个拷贝、10000倍稀释液的拷贝数为27个拷贝。由此可以得出,本实施例1得到的引物对于检测L24-2位点的TCR-T细胞药的灵敏度能达到27个拷贝。即,在实际应用中,引物在患者外周血样的基因组DNA中最低可以检测到27个拷贝的TCR基因片段。
将L24-49位点的引物按照本实施例3的灵敏度的检测方法检测相对应位点的TCR-T细胞药,所得到的灵敏度均能达到27个拷贝。
实施例4
按照SEQ ID NO.12所示的质粒载体设置L24-2和L24-49位点对应的TCR基因片段,得到SEQ ID NO.13~14所示的基因片段,所述L24-2位点对应SEQ ID NO.13,L24-49位点对应SEQ ID NO.14。利用这两个位点设计引物来检测相应位点的TCR-T细胞药物,同样能得到实施例1~3所述引物和效果。
由以上实施例可知,本发明提供了一种特异性识别IYVLVMLVL肽段的荧光PCR引物组及其应用。本申请的引物组是根据HLAA24分型,EBV病毒抗原肽段IYVLVMLVL的TCR设计的。将本申请的引物组制备成荧光定量PCR试剂盒,追踪患者体内TCR-T细胞药物时,能特异的检测到27个拷贝数的TCR-T细胞药物。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
序列表
<110> 重庆天科雅生物科技有限公司
<120> 一种特异性识别IYVLVMLVL肽段的荧光PCR引物组及其应用
<150> 202110687266.8
<151> 2021-06-21
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agcgtgtaca gcagcggcaa cgaggacggc agattcacag cccagctgaa cagagccagc 3120
cagtacatca gcctgctgat ccgggatagc aagctgagcg atagcgccac ctacctgtgt 3180
gtcgtgcggg gctacaactt caacaagttc tacttcggca gcggcaccaa gctgaacgtg 3240
aagcctgaca ttcagaatcc cgaacccgcc gtgtaccagc tgaaggaccc taggagccag 3300
gactctaccc tctgcctgtt caccgacttc gacagccaga tcaacgtgcc caagacaatg 3360
gagagcggca ccttcatcac cgacaagacc gtgctggaca tgaaggctat ggacagcaag 3420
agcaacggag ccatcgcttg gagcaaccag accagcttca cttgccagga catcttcaag 3480
gagaccaacg cttgctaccc ctctagcgac gtgccttgcg acgccacact gacagagaag 3540
agcttcgaga ccgacatgaa cctgaacttc cagaacctga gcgtgatggg cctgagaatc 3600
ctgctgctga aggtggccgg attcaacctg ctgatgaccc tgaggctgtg gtcttcccga 3660
gctaaacgag gctcaggcgc gacgaacttt agtttgctga agcaagctgg tgacgttgag 3720
gaaaatccgg gacccatggc cacaggcagc agaacatctc tgctgctggc cttcggactg 3780
ctgtgcctgc cttgtctgca agagggatct gccggtgctg tggtgtctca gcacccttct 3840
tgggtcatct gcaagtccgg caccagcgtg aagatcgagt gcagaagcct ggacttccag 3900
gccaccacca tgttctggta cagacagttc cccaagcaga gcctgatgct gatggccacc 3960
tctaacgagg gcagcaaggc cacatatgag cagggcgtcg agaaggacaa gttcctgatc 4020
aaccacgcca gcctgacact gagcaccctg acagtgacaa gcgcccatcc tgaggacagc 4080
agcttctaca tctgcagcgc cagaggaccc agaatcaccg acagcagcac cgacacacag 4140
tacttcggcc ctggcaccag actgacagtt ctggaggacc tacgtaacgt gacccctccc 4200
aaggtgtccc tgttcgagcc tagcaaggcc gagatcgcca acaagcagaa ggccacactc 4260
gtctgcctgg ctagaggctt cttcccagac cacgtggagc tgtcttggtg ggtgaacggc 4320
aaggaggtgc actcaggagt gtctaccgac cctcaggcct acaaggagag caactacagc 4380
tactgcctgt cctccagact cagggtctgc gccacctttt ggcacaaccc tcggaaccac 4440
ttccgctgtc aggtccagtt ccacggcctg tccgaggaag acaagtggcc agagggctct 4500
cctaagccag tgacacagaa catcagcgcc gaggcttggg gaagagccga ttgcggaatc 4560
accagcgcct cttaccagca gggagtgctg tcagctacca tcctgtacga gatcctgctg 4620
ggcaaggcca cactgtacgc agtgctggtg tccactctgg tcgtgatggc tatggtgaag 4680
cggaagaaca gctaggaatt cgagcatctt accgccattt attcccatat ttgttctgtt 4740
tttcttgatt tgggtataca tttaaatgtt aataaaacaa aatggtgggg caatcattta 4800
cattttatgg gatatgtaat tactagttca ggtgtattgc cacaagacaa acatgttaag 4860
aaactttccc gttatttacg ctctgttcct gttaatcaac ctctggatta caaaatttgt 4920
gaaagattga ctgatattct taactatgtt gctcctttta cgctgtgtgg atatgctgct 4980
ttaatgcctc tgtatcatgc tattgcttcc cgtacggctt tcgttttctc ctccttgtat 5040
aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtccgtca acgtggcgtg 5100
gtgtgctctg tgtttgctga cgcaaccccc actggctggg gcattgccac cacctgtcaa 5160
ctcctttctg ggactttcgc tttccccctc ccgatcgcca cggcagaact catcgccgcc 5220
tgccttgccc gctgctggac aggggctagg ttgctgggca ctgataattc cgtggtgttg 5280
tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccaactg gatcctgcgc 5340
gggacgtcct tctgctacgt cccttcggct ctcaatccag cggacctccc ttcccgaggc 5400
cttctgccgg ttctgcggcc tctcccgcgt cttcgctttc ggcctccgac gagtcggatc 5460
tccctttggg ccgcctcccc gcctgtttcg cctcggcgtc cggtccgtgt tgcttggtcg 5520
tcacctgtgc agaattgcga accatggatt ccaccgtgaa ctttgtctcc tggcatgcaa 5580
atcgtcaact tggcatgcca agaataattc ggatccaagc ttaggcctgc tcgctttctt 5640
gctgtcccat ttctattaaa ggttcctttg ttccctaagt ccaactacta aactggggga 5700
tattatgaag ggccttgagc atctggattc tgcctagcgc taagcttaac acgagccata 5760
gatagaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg 5820
taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg 5880
agaatagaga agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca 5940
ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagtt ggaacagcag 6000
aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa 6060
cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc 6120
cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg 6180
cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc 6240
tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg ttctcaataa 6300
accctcttgc agttgcatcc gactcgtggt ctcgctgttc cttgggaggg tctcctctga 6360
gtgattgact gcccacctcg ggggtctttc attctcgagc agcttggcgt aatcatggtc 6420
atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 6480
aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 6540
gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 6600
ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 6660
ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 6720
acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 6780
aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 6840
tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 6900
aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 6960
gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 7020
acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 7080
accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 7140
ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 7200
gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 7260
aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagt 7314
<210> 6
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
agcagagcct gatgctgatg 20
<210> 7
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
gtattggcct ctctctcgcc 20
<210> 8
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
aaggccacat atgagcaggg 20
<210> 9
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
agtactgtgt gtcggtgctg 20
<210> 10
<211> 7263
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 60
gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 120
gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 180
aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 240
atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 300
gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 360
acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 420
ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 480
tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 540
ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 600
ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 660
atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 720
taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 780
catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 840
atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 900
acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 960
aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 1020
ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 1080
cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 1140
atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 1200
ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 1260
ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt 1320
tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 1380
ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 1440
gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 1500
agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag 1560
gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc 1620
gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc 1680
agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattagtac tctagcttaa 1740
gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa 1800
ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc 1860
ctgccccggc tcagggccaa gaacagttgg aacagcagaa tatgggccaa acaggatatc 1920
tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc 1980
ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa 2040
atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc 2100
ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc 2160
gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg 2220
aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg 2280
gggtctttca tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc 2340
cccccgccgg gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt 2400
gtttgtgccg gcatctaatg tttgcgcctg cgtctgtact agttggctaa ctagatctgt 2460
atctggcggt cccgcggaag aactgacgag ttcgtattcc cggccgcagc ccctgggaga 2520
cgtcccagcg gcctcggggg cccgttttgt ggcccattct gtatcagtta acctacccga 2580
gtcggacttt ttggagctcc gccactgtcc gaggggtacg tggctttgtt gggggacgag 2640
agacagagac acttcccgcc cccgtctgaa tttttgcttt cggttttacg ccgaaaccgc 2700
gccgcgcgtc ttgtctgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg 2760
tatttgtctg aaaattagct cgacaaagtt aagtaatagt ccctctctcc aagctcactt 2820
acaggcggcc gcgccaccat gatcagcctg agagtgctgc tggtcatcct gtggctgcag 2880
ctgtcttggg tctggtccca gcggaaagag gtggaacagg accccggacc tttcaatgtg 2940
cctgaaggcg ccaccgtggc cttcaactgc acctacagca atagcgccag ccagagcttc 3000
ttctggtaca gacaggactg ccggaaagaa cccaagctgc tgatgagcgt gtacagcagc 3060
ggcaacgagg acggcagatt cacagcccag ctgaacagag ccagccagta catcagcctg 3120
ctgatccggg atagcaagct gagcgatagc gccacctacc tgtgtgtcgt gcggggctac 3180
aacttcaaca agttctactt cggcagcggc accaagctga acgtgaagcc tgacattcag 3240
aatcccgaac ccgccgtgta ccagctgaag gaccctagga gccaggactc taccctctgc 3300
ctgttcaccg acttcgacag ccagatcaac gtgcccaaga caatggagag cggcaccttc 3360
atcaccgaca agaccgtgct ggacatgaag gctatggaca gcaagagcaa cggagccatc 3420
gcttggagca accagaccag cttcacttgc caggacatct tcaaggagac caacgcttgc 3480
tacccctcta gcgacgtgcc ttgcgacgcc acactgacag agaagagctt cgagaccgac 3540
atgaacctga acttccagaa cctgagcgtg atgggcctga gaatcctgct gctgaaggtg 3600
gccggattca acctgctgat gaccctgagg ctgtggtctt cccgagctaa acgaggctca 3660
ggcgcgacga actttagttt gctgaagcaa gctggggatg tagaggaaaa tccgggtccc 3720
atgctgctgc tcctgctgct tctcggacct ggatctggac tgggagctgt ggtgtctcag 3780
cacccctcta gagtgatctg caagagcggc accagcgtga agatcgagtg cagaagcctg 3840
gacttccagg ccaccaccat gttctggtac agacagttcc ccaagcagag cctgatgctg 3900
atggccacct ctaacgaggg cagcaaggcc acatatgagc agggcgtcga gaaggacaag 3960
ttcctgatca accacgccag cctgacactg agcaccctga cagtgacaag cgcccatcct 4020
gaggacagca gcttctacat ctgcgccagt agaggccctg gcgagagaga ggccaatacc 4080
gacacacagt acttcggccc tggcaccaga ctgctggttc ttgaggacct acgtaacgtg 4140
acccctccca aggtgtccct gttcgagcct agcaaggccg agatcgccaa caagcagaag 4200
gccacactcg tctgcctggc tagaggcttc ttcccagacc acgtggagct gtcttggtgg 4260
gtgaacggca aggaggtgca ctcaggagtg tctaccgacc ctcaggccta caaggagagc 4320
aactacagct actgcctgtc ctccagactc agggtctgcg ccaccttttg gcacaaccct 4380
cggaaccact tccgctgtca ggtccagttc cacggcctgt ccgaggaaga caagtggcca 4440
gagggctctc ctaagccagt gacacagaac atcagcgccg aggcttgggg aagagccgat 4500
tgcggaatca ccagcgcctc ttaccagcag ggagtgctgt cagctaccat cctgtacgag 4560
atcctgctgg gcaaggccac actgtacgca gtgctggtgt ccactctggt cgtgatggct 4620
atggtgaagc ggaagaacag ctaggaattc gagcatctta ccgccattta ttcccatatt 4680
tgttctgttt ttcttgattt gggtatacat ttaaatgtta ataaaacaaa atggtggggc 4740
aatcatttac attttatggg atatgtaatt actagttcag gtgtattgcc acaagacaaa 4800
catgttaaga aactttcccg ttatttacgc tctgttcctg ttaatcaacc tctggattac 4860
aaaatttgtg aaagattgac tgatattctt aactatgttg ctccttttac gctgtgtgga 4920
tatgctgctt taatgcctct gtatcatgct attgcttccc gtacggcttt cgttttctcc 4980
tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtccgtcaa 5040
cgtggcgtgg tgtgctctgt gtttgctgac gcaaccccca ctggctgggg cattgccacc 5100
acctgtcaac tcctttctgg gactttcgct ttccccctcc cgatcgccac ggcagaactc 5160
atcgccgcct gccttgcccg ctgctggaca ggggctaggt tgctgggcac tgataattcc 5220
gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccaactgg 5280
atcctgcgcg ggacgtcctt ctgctacgtc ccttcggctc tcaatccagc ggacctccct 5340
tcccgaggcc ttctgccggt tctgcggcct ctcccgcgtc ttcgctttcg gcctccgacg 5400
agtcggatct ccctttgggc cgcctccccg cctgtttcgc ctcggcgtcc ggtccgtgtt 5460
gcttggtcgt cacctgtgca gaattgcgaa ccatggattc caccgtgaac tttgtctcct 5520
ggcatgcaaa tcgtcaactt ggcatgccaa gaataattcg gatccaagct taggcctgct 5580
cgctttcttg ctgtcccatt tctattaaag gttcctttgt tccctaagtc caactactaa 5640
actgggggat attatgaagg gccttgagca tctggattct gcctagcgct aagcttaaca 5700
cgagccatag atagaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga 5760
ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat 5820
acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg 5880
ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagttg 5940
gaacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag 6000
ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc 6060
agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca 6120
atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc 6180
cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt 6240
tctcaataaa ccctcttgca gttgcatccg actcgtggtc tcgctgttcc ttgggagggt 6300
ctcctctgag tgattgactg cccacctcgg gggtctttca ttctcgagca gcttggcgta 6360
atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 6420
acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 6480
aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 6540
atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 6600
gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 6660
ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 6720
aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 6780
ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 6840
aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 6900
gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 6960
tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 7020
tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 7080
gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 7140
cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 7200
cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 7260
agt 7263
<210> 11
<211> 7314
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 60
gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 120
gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 180
aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 240
atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 300
gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 360
acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 420
ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 480
tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 540
ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 600
ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 660
atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 720
taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 780
catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 840
atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 900
acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 960
aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 1020
ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 1080
cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 1140
atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 1200
ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 1260
ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt 1320
tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 1380
ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 1440
gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 1500
agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag 1560
gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc 1620
gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc 1680
agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattagtac tctagcttaa 1740
gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa 1800
ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc 1860
ctgccccggc tcagggccaa gaacagttgg aacagcagaa tatgggccaa acaggatatc 1920
tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc 1980
ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa 2040
atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc 2100
ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc 2160
gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg 2220
aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg 2280
gggtctttca tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc 2340
cccccgccgg gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt 2400
gtttgtgccg gcatctaatg tttgcgcctg cgtctgtact agttggctaa ctagatctgt 2460
atctggcggt cccgcggaag aactgacgag ttcgtattcc cggccgcagc ccctgggaga 2520
cgtcccagcg gcctcggggg cccgttttgt ggcccattct gtatcagtta acctacccga 2580
gtcggacttt ttggagctcc gccactgtcc gaggggtacg tggctttgtt gggggacgag 2640
agacagagac acttcccgcc cccgtctgaa tttttgcttt cggttttacg ccgaaaccgc 2700
gccgcgcgtc ttgtctgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg 2760
tatttgtctg aaaattagct cgacaaagtt aagtaatagt ccctctctcc aagctcactt 2820
acaggcggcc gcgccaccat ggccacaggc agcagaacat ctctgctgct ggccttcgga 2880
ctgctgtgtc tgccttggct gcaagaagcc agcgccagaa aagaggtgga acaggacccc 2940
ggacctttca atgtgcctga aggcgccacc gtggccttca actgcaccta cagcaatagc 3000
gccagccaga gcttcttctg gtacagacag gactgccgga aagaacccaa gctgctgatg 3060
agcgtgtaca gcagcggcaa cgaggacggc agattcacag cccagctgaa cagagccagc 3120
cagtacatca gcctgctgat ccgggatagc aagctgagcg atagcgccac ctacctgtgt 3180
gtcgtgcggg gctacaactt caacaagttc tacttcggca gcggcaccaa gctgaacgtg 3240
aagcctgaca ttcagaatcc cgaacccgcc gtgtaccagc tgaaggaccc taggagccag 3300
gactctaccc tctgcctgtt caccgacttc gacagccaga tcaacgtgcc caagacaatg 3360
gagagcggca ccttcatcac cgacaagacc gtgctggaca tgaaggctat ggacagcaag 3420
agcaacggag ccatcgcttg gagcaaccag accagcttca cttgccagga catcttcaag 3480
gagaccaacg cttgctaccc ctctagcgac gtgccttgcg acgccacact gacagagaag 3540
agcttcgaga ccgacatgaa cctgaacttc cagaacctga gcgtgatggg cctgagaatc 3600
ctgctgctga aggtggccgg attcaacctg ctgatgaccc tgaggctgtg gtcttcccga 3660
gctaaacgag gctcaggcgc gacgaacttt agtttgctga agcaagctgg ggatgtagag 3720
gaaaatccgg gtcccatggc cacaggcagc agaacatctc tgctgctggc cttcggactg 3780
ctgtgcctgc cttgtctgca agagggatct gccggtgctg tggtgtctca gcacccttct 3840
tgggtcatct gcaagtccgg caccagcgtg aagatcgagt gcagaagcct ggacttccag 3900
gccaccacca tgttctggta cagacagttc cccaagcaga gcctgatgct gatggccacc 3960
tctaacgagg gcagcaaggc cacatatgag cagggcgtcg agaaggacaa gttcctgatc 4020
aaccacgcca gcctgacact gagcaccctg acagtgacaa gcgcccatcc tgaggacagc 4080
agcttctaca tctgcagcgc cagaggaccc agaatcaccg acagcagcac cgacacacag 4140
tacttcggcc ctggcaccag actgacagtt ctggaggacc tacgtaacgt gacccctccc 4200
aaggtgtccc tgttcgagcc tagcaaggcc gagatcgcca acaagcagaa ggccacactc 4260
gtctgcctgg ctagaggctt cttcccagac cacgtggagc tgtcttggtg ggtgaacggc 4320
aaggaggtgc actcaggagt gtctaccgac cctcaggcct acaaggagag caactacagc 4380
tactgcctgt cctccagact cagggtctgc gccacctttt ggcacaaccc tcggaaccac 4440
ttccgctgtc aggtccagtt ccacggcctg tccgaggaag acaagtggcc agagggctct 4500
cctaagccag tgacacagaa catcagcgcc gaggcttggg gaagagccga ttgcggaatc 4560
accagcgcct cttaccagca gggagtgctg tcagctacca tcctgtacga gatcctgctg 4620
ggcaaggcca cactgtacgc agtgctggtg tccactctgg tcgtgatggc tatggtgaag 4680
cggaagaaca gctaggaatt cgagcatctt accgccattt attcccatat ttgttctgtt 4740
tttcttgatt tgggtataca tttaaatgtt aataaaacaa aatggtgggg caatcattta 4800
cattttatgg gatatgtaat tactagttca ggtgtattgc cacaagacaa acatgttaag 4860
aaactttccc gttatttacg ctctgttcct gttaatcaac ctctggatta caaaatttgt 4920
gaaagattga ctgatattct taactatgtt gctcctttta cgctgtgtgg atatgctgct 4980
ttaatgcctc tgtatcatgc tattgcttcc cgtacggctt tcgttttctc ctccttgtat 5040
aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtccgtca acgtggcgtg 5100
gtgtgctctg tgtttgctga cgcaaccccc actggctggg gcattgccac cacctgtcaa 5160
ctcctttctg ggactttcgc tttccccctc ccgatcgcca cggcagaact catcgccgcc 5220
tgccttgccc gctgctggac aggggctagg ttgctgggca ctgataattc cgtggtgttg 5280
tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccaactg gatcctgcgc 5340
gggacgtcct tctgctacgt cccttcggct ctcaatccag cggacctccc ttcccgaggc 5400
cttctgccgg ttctgcggcc tctcccgcgt cttcgctttc ggcctccgac gagtcggatc 5460
tccctttggg ccgcctcccc gcctgtttcg cctcggcgtc cggtccgtgt tgcttggtcg 5520
tcacctgtgc agaattgcga accatggatt ccaccgtgaa ctttgtctcc tggcatgcaa 5580
atcgtcaact tggcatgcca agaataattc ggatccaagc ttaggcctgc tcgctttctt 5640
gctgtcccat ttctattaaa ggttcctttg ttccctaagt ccaactacta aactggggga 5700
tattatgaag ggccttgagc atctggattc tgcctagcgc taagcttaac acgagccata 5760
gatagaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg 5820
taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg 5880
agaatagaga agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca 5940
ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagtt ggaacagcag 6000
aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa 6060
cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc 6120
cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg 6180
cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc 6240
tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg ttctcaataa 6300
accctcttgc agttgcatcc gactcgtggt ctcgctgttc cttgggaggg tctcctctga 6360
gtgattgact gcccacctcg ggggtctttc attctcgagc agcttggcgt aatcatggtc 6420
atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 6480
aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 6540
gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 6600
ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 6660
ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 6720
acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 6780
aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 6840
tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 6900
aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 6960
gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 7020
acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 7080
accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 7140
ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 7200
gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 7260
aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagt 7314
<210> 12
<211> 5979
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
gaggacctac gtaacgtgac ccctcccaag gtgtccctgt tcgagcctag caaggccgag 60
atcgccaaca agcagaaggc cacactcgtc tgcctggcta gaggcttctt cccagaccac 120
gtggagctgt cttggtgggt gaacggcaag gaggtgcact caggagtgtc taccgaccct 180
caggcctaca aggagagcaa ctacagctac tgcctgtcct ccagactcag ggtctgcgcc 240
accttttggc acaaccctcg gaaccacttc cgctgtcagg tccagttcca cggcctgtcc 300
gaggaagaca agtggccaga gggctctcct aagccagtga cacagaacat cagcgccgag 360
gcttggggaa gagccgattg cggaatcacc agcgcctctt accagcaggg agtgctgtca 420
gctaccatcc tgtacgagat cctgctgggc aaggccacac tgtacgcagt gctggtgtcc 480
actctggtcg tgatggctat ggtgaagcgg aagaacagct aggaattcga gcatcttacc 540
gccatttatt cccatatttg ttctgttttt cttgatttgg gtatacattt aaatgttaat 600
aaaacaaaat ggtggggcaa tcatttacat tttatgggat atgtaattac tagttcaggt 660
gtattgccac aagacaaaca tgttaagaaa ctttcccgtt atttacgctc tgttcctgtt 720
aatcaacctc tggattacaa aatttgtgaa agattgactg atattcttaa ctatgttgct 780
ccttttacgc tgtgtggata tgctgcttta atgcctctgt atcatgctat tgcttcccgt 840
acggctttcg ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 900
tggcccgttg tccgtcaacg tggcgtggtg tgctctgtgt ttgctgacgc aacccccact 960
ggctggggca ttgccaccac ctgtcaactc ctttctggga ctttcgcttt ccccctcccg 1020
atcgccacgg cagaactcat cgccgcctgc cttgcccgct gctggacagg ggctaggttg 1080
ctgggcactg ataattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc 1140
gcctgtgttg ccaactggat cctgcgcggg acgtccttct gctacgtccc ttcggctctc 1200
aatccagcgg acctcccttc ccgaggcctt ctgccggttc tgcggcctct cccgcgtctt 1260
cgctttcggc ctccgacgag tcggatctcc ctttgggccg cctccccgcc tgtttcgcct 1320
cggcgtccgg tccgtgttgc ttggtcgtca cctgtgcaga attgcgaacc atggattcca 1380
ccgtgaactt tgtctcctgg catgcaaatc gtcaacttgg catgccaaga ataattcgga 1440
tccaagctta ggcctgctcg ctttcttgct gtcccatttc tattaaaggt tcctttgttc 1500
cctaagtcca actactaaac tgggggatat tatgaagggc cttgagcatc tggattctgc 1560
ctagcgctaa gcttaacacg agccatagat agaataaaag attttattta gtctccagaa 1620
aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt 1680
ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca 1740
gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct 1800
cagggccaag aacagttgga acagcagaat atgggccaaa caggatatct gtggtaagca 1860
gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc 1920
agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg 1980
ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc 2040
gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg 2100
tcgcccgggt acccgtgttc tcaataaacc ctcttgcagt tgcatccgac tcgtggtctc 2160
gctgttcctt gggagggtct cctctgagtg attgactgcc cacctcgggg gtctttcatt 2220
ctcgagcagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 2280
cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 2340
agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 2400
gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 2460
gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 2520
ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 2580
aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 2640
ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 2700
gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 2760
cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 2820
gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 2880
tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 2940
cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 3000
cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 3060
gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 3120
agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 3180
cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 3240
tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 3300
tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 3360
ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 3420
cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 3480
cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 3540
accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 3600
ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 3660
ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 3720
tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 3780
acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 3840
tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 3900
actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 3960
ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 4020
aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 4080
ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 4140
cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 4200
aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 4260
actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 4320
cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 4380
ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 4440
taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 4500
acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 4560
agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 4620
atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 4680
aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 4740
gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 4800
gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 4860
tgaattagta ctctagctta agtaacgcca ttttgcaagg catggaaaat acataactga 4920
gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag 4980
gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagttg gaacagcaga 5040
atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac 5100
agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc 5160
agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc 5220
ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct 5280
cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtat tcccaataaa 5340
gcctcttgct gtttgcatcc gaatcgtgga ctcgctgatc cttgggaggg tctcctcaga 5400
ttgattgact gcccacctcg ggggtctttc atttggaggt tccaccgaga tttggagacc 5460
cctgcccagg gaccaccgac ccccccgccg ggaggtaagc tggccagcgg tcgtttcgtg 5520
tctgtctctg tctttgtgcg tgtttgtgcc ggcatctaat gtttgcgcct gcgtctgtac 5580
tagttggcta actagatctg tatctggcgg tcccgcggaa gaactgacga gttcgtattc 5640
ccggccgcag cccctgggag acgtcccagc ggcctcgggg gcccgttttg tggcccattc 5700
tgtatcagtt aacctacccg agtcggactt tttggagctc cgccactgtc cgaggggtac 5760
gtggctttgt tgggggacga gagacagaga cacttcccgc ccccgtctga atttttgctt 5820
tcggttttac gccgaaaccg cgccgcgcgt cttgtctgct gcagcatcgt tctgtgttgt 5880
ctctgtctga ctgtgtttct gtatttgtct gaaaattagc tcgacaaagt taagtaatag 5940
tccctctctc caagctcact tacaggcggc cgcgccacc 5979
<210> 13
<211> 7263
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 60
gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 120
gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 180
aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 240
atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 300
gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 360
acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 420
ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 480
tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 540
ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 600
ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 660
atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 720
taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 780
catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 840
atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 900
acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 960
aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 1020
ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 1080
cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 1140
atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 1200
ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 1260
ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt 1320
tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 1380
ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 1440
gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 1500
agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag 1560
gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc 1620
gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc 1680
agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattagtac tctagcttaa 1740
gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa 1800
ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc 1860
ctgccccggc tcagggccaa gaacagttgg aacagcagaa tatgggccaa acaggatatc 1920
tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc 1980
ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa 2040
atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc 2100
ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc 2160
gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg 2220
aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg 2280
gggtctttca tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc 2340
cccccgccgg gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt 2400
gtttgtgccg gcatctaatg tttgcgcctg cgtctgtact agttggctaa ctagatctgt 2460
atctggcggt cccgcggaag aactgacgag ttcgtattcc cggccgcagc ccctgggaga 2520
cgtcccagcg gcctcggggg cccgttttgt ggcccattct gtatcagtta acctacccga 2580
gtcggacttt ttggagctcc gccactgtcc gaggggtacg tggctttgtt gggggacgag 2640
agacagagac acttcccgcc cccgtctgaa tttttgcttt cggttttacg ccgaaaccgc 2700
gccgcgcgtc ttgtctgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg 2760
tatttgtctg aaaattagct cgacaaagtt aagtaatagt ccctctctcc aagctcactt 2820
acaggcggcc gcgccaccat gatcagcctg agagtgctgc tggtcatcct gtggctgcag 2880
ctgtcttggg tctggtccca gcggaaagag gtggaacagg accccggacc tttcaatgtg 2940
cctgaaggcg ccaccgtggc cttcaactgc acctacagca atagcgccag ccagagcttc 3000
ttctggtaca gacaggactg ccggaaagaa cccaagctgc tgatgagcgt gtacagcagc 3060
ggcaacgagg acggcagatt cacagcccag ctgaacagag ccagccagta catcagcctg 3120
ctgatccggg atagcaagct gagcgatagc gccacctacc tgtgtgtcgt gcggggctac 3180
aacttcaaca agttctactt cggcagcggc accaagctga acgtgaagcc tgacattcag 3240
aatcccgaac ccgccgtgta ccagctgaag gaccctagga gccaggactc taccctctgc 3300
ctgttcaccg acttcgacag ccagatcaac gtgcccaaga caatggagag cggcaccttc 3360
atcaccgaca agaccgtgct ggacatgaag gctatggaca gcaagagcaa cggagccatc 3420
gcttggagca accagaccag cttcacttgc caggacatct tcaaggagac caacgcttgc 3480
tacccctcta gcgacgtgcc ttgcgacgcc acactgacag agaagagctt cgagaccgac 3540
atgaacctga acttccagaa cctgagcgtg atgggcctga gaatcctgct gctgaaggtg 3600
gccggattca acctgctgat gaccctgagg ctgtggtctt cccgagctaa acgaggctca 3660
ggcgcgacga actttagttt gctgaagcaa gctggggatg tagaggaaaa tccgggtccc 3720
atgctgctgc tcctgctgct tctcggacct ggatctggac tgggagctgt ggtgtctcag 3780
cacccctcta gagtgatctg caagagcggc accagcgtga agatcgagtg cagaagcctg 3840
gacttccagg ccaccaccat gttctggtac agacagttcc ccaagcagag cctgatgctg 3900
atggccacct ctaacgaggg cagcaaggcc acatatgagc agggcgtcga gaaggacaag 3960
ttcctgatca accacgccag cctgacactg agcaccctga cagtgacaag cgcccatcct 4020
gaggacagca gcttctacat ctgcgccagt agaggccctg gcgagagaga ggccaatacc 4080
gacacacagt acttcggccc tggcaccaga ctgctggttc ttgaggacct acgtaacgtg 4140
acccctccca aggtgtccct gttcgagcct agcaaggccg agatcgccaa caagcagaag 4200
gccacactcg tctgcctggc tagaggcttc ttcccagacc acgtggagct gtcttggtgg 4260
gtgaacggca aggaggtgca ctcaggagtg tctaccgacc ctcaggccta caaggagagc 4320
aactacagct actgcctgtc ctccagactc agggtctgcg ccaccttttg gcacaaccct 4380
cggaaccact tccgctgtca ggtccagttc cacggcctgt ccgaggaaga caagtggcca 4440
gagggctctc ctaagccagt gacacagaac atcagcgccg aggcttgggg aagagccgat 4500
tgcggaatca ccagcgcctc ttaccagcag ggagtgctgt cagctaccat cctgtacgag 4560
atcctgctgg gcaaggccac actgtacgca gtgctggtgt ccactctggt cgtgatggct 4620
atggtgaagc ggaagaacag ctaggaattc gagcatctta ccgccattta ttcccatatt 4680
tgttctgttt ttcttgattt gggtatacat ttaaatgtta ataaaacaaa atggtggggc 4740
aatcatttac attttatggg atatgtaatt actagttcag gtgtattgcc acaagacaaa 4800
catgttaaga aactttcccg ttatttacgc tctgttcctg ttaatcaacc tctggattac 4860
aaaatttgtg aaagattgac tgatattctt aactatgttg ctccttttac gctgtgtgga 4920
tatgctgctt taatgcctct gtatcatgct attgcttccc gtacggcttt cgttttctcc 4980
tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtccgtcaa 5040
cgtggcgtgg tgtgctctgt gtttgctgac gcaaccccca ctggctgggg cattgccacc 5100
acctgtcaac tcctttctgg gactttcgct ttccccctcc cgatcgccac ggcagaactc 5160
atcgccgcct gccttgcccg ctgctggaca ggggctaggt tgctgggcac tgataattcc 5220
gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccaactgg 5280
atcctgcgcg ggacgtcctt ctgctacgtc ccttcggctc tcaatccagc ggacctccct 5340
tcccgaggcc ttctgccggt tctgcggcct ctcccgcgtc ttcgctttcg gcctccgacg 5400
agtcggatct ccctttgggc cgcctccccg cctgtttcgc ctcggcgtcc ggtccgtgtt 5460
gcttggtcgt cacctgtgca gaattgcgaa ccatggattc caccgtgaac tttgtctcct 5520
ggcatgcaaa tcgtcaactt ggcatgccaa gaataattcg gatccaagct taggcctgct 5580
cgctttcttg ctgtcccatt tctattaaag gttcctttgt tccctaagtc caactactaa 5640
actgggggat attatgaagg gccttgagca tctggattct gcctagcgct aagcttaaca 5700
cgagccatag atagaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga 5760
ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat 5820
acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg 5880
ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagttg 5940
gaacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag 6000
ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc 6060
agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca 6120
atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc 6180
cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt 6240
tctcaataaa ccctcttgca gttgcatccg actcgtggtc tcgctgttcc ttgggagggt 6300
ctcctctgag tgattgactg cccacctcgg gggtctttca ttctcgagca gcttggcgta 6360
atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 6420
acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 6480
aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 6540
atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 6600
gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 6660
ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 6720
aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 6780
ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 6840
aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 6900
gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 6960
tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 7020
tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 7080
gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 7140
cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 7200
cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 7260
agt 7263
<210> 14
<211> 7314
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 60
gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 120
gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 180
aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 240
atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 300
gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 360
acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 420
ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 480
tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 540
ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 600
ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 660
atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 720
taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 780
catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 840
atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 900
acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 960
aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 1020
ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 1080
cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 1140
atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 1200
ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 1260
ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt 1320
tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 1380
ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 1440
gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 1500
agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag 1560
gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc 1620
gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc 1680
agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattagtac tctagcttaa 1740
gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa 1800
ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc 1860
ctgccccggc tcagggccaa gaacagttgg aacagcagaa tatgggccaa acaggatatc 1920
tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc 1980
ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa 2040
atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc 2100
ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc 2160
gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg 2220
aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg 2280
gggtctttca tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc 2340
cccccgccgg gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt 2400
gtttgtgccg gcatctaatg tttgcgcctg cgtctgtact agttggctaa ctagatctgt 2460
atctggcggt cccgcggaag aactgacgag ttcgtattcc cggccgcagc ccctgggaga 2520
cgtcccagcg gcctcggggg cccgttttgt ggcccattct gtatcagtta acctacccga 2580
gtcggacttt ttggagctcc gccactgtcc gaggggtacg tggctttgtt gggggacgag 2640
agacagagac acttcccgcc cccgtctgaa tttttgcttt cggttttacg ccgaaaccgc 2700
gccgcgcgtc ttgtctgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg 2760
tatttgtctg aaaattagct cgacaaagtt aagtaatagt ccctctctcc aagctcactt 2820
acaggcggcc gcgccaccat ggccacaggc agcagaacat ctctgctgct ggccttcgga 2880
ctgctgtgtc tgccttggct gcaagaagcc agcgccagaa aagaggtgga acaggacccc 2940
ggacctttca atgtgcctga aggcgccacc gtggccttca actgcaccta cagcaatagc 3000
gccagccaga gcttcttctg gtacagacag gactgccgga aagaacccaa gctgctgatg 3060
agcgtgtaca gcagcggcaa cgaggacggc agattcacag cccagctgaa cagagccagc 3120
cagtacatca gcctgctgat ccgggatagc aagctgagcg atagcgccac ctacctgtgt 3180
gtcgtgcggg gctacaactt caacaagttc tacttcggca gcggcaccaa gctgaacgtg 3240
aagcctgaca ttcagaatcc cgaacccgcc gtgtaccagc tgaaggaccc taggagccag 3300
gactctaccc tctgcctgtt caccgacttc gacagccaga tcaacgtgcc caagacaatg 3360
gagagcggca ccttcatcac cgacaagacc gtgctggaca tgaaggctat ggacagcaag 3420
agcaacggag ccatcgcttg gagcaaccag accagcttca cttgccagga catcttcaag 3480
gagaccaacg cttgctaccc ctctagcgac gtgccttgcg acgccacact gacagagaag 3540
agcttcgaga ccgacatgaa cctgaacttc cagaacctga gcgtgatggg cctgagaatc 3600
ctgctgctga aggtggccgg attcaacctg ctgatgaccc tgaggctgtg gtcttcccga 3660
gctaaacgag gctcaggcgc gacgaacttt agtttgctga agcaagctgg ggatgtagag 3720
gaaaatccgg gtcccatggc cacaggcagc agaacatctc tgctgctggc cttcggactg 3780
ctgtgcctgc cttgtctgca agagggatct gccggtgctg tggtgtctca gcacccttct 3840
tgggtcatct gcaagtccgg caccagcgtg aagatcgagt gcagaagcct ggacttccag 3900
gccaccacca tgttctggta cagacagttc cccaagcaga gcctgatgct gatggccacc 3960
tctaacgagg gcagcaaggc cacatatgag cagggcgtcg agaaggacaa gttcctgatc 4020
aaccacgcca gcctgacact gagcaccctg acagtgacaa gcgcccatcc tgaggacagc 4080
agcttctaca tctgcagcgc cagaggaccc agaatcaccg acagcagcac cgacacacag 4140
tacttcggcc ctggcaccag actgacagtt ctggaggacc tacgtaacgt gacccctccc 4200
aaggtgtccc tgttcgagcc tagcaaggcc gagatcgcca acaagcagaa ggccacactc 4260
gtctgcctgg ctagaggctt cttcccagac cacgtggagc tgtcttggtg ggtgaacggc 4320
aaggaggtgc actcaggagt gtctaccgac cctcaggcct acaaggagag caactacagc 4380
tactgcctgt cctccagact cagggtctgc gccacctttt ggcacaaccc tcggaaccac 4440
ttccgctgtc aggtccagtt ccacggcctg tccgaggaag acaagtggcc agagggctct 4500
cctaagccag tgacacagaa catcagcgcc gaggcttggg gaagagccga ttgcggaatc 4560
accagcgcct cttaccagca gggagtgctg tcagctacca tcctgtacga gatcctgctg 4620
ggcaaggcca cactgtacgc agtgctggtg tccactctgg tcgtgatggc tatggtgaag 4680
cggaagaaca gctaggaatt cgagcatctt accgccattt attcccatat ttgttctgtt 4740
tttcttgatt tgggtataca tttaaatgtt aataaaacaa aatggtgggg caatcattta 4800
cattttatgg gatatgtaat tactagttca ggtgtattgc cacaagacaa acatgttaag 4860
aaactttccc gttatttacg ctctgttcct gttaatcaac ctctggatta caaaatttgt 4920
gaaagattga ctgatattct taactatgtt gctcctttta cgctgtgtgg atatgctgct 4980
ttaatgcctc tgtatcatgc tattgcttcc cgtacggctt tcgttttctc ctccttgtat 5040
aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtccgtca acgtggcgtg 5100
gtgtgctctg tgtttgctga cgcaaccccc actggctggg gcattgccac cacctgtcaa 5160
ctcctttctg ggactttcgc tttccccctc ccgatcgcca cggcagaact catcgccgcc 5220
tgccttgccc gctgctggac aggggctagg ttgctgggca ctgataattc cgtggtgttg 5280
tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccaactg gatcctgcgc 5340
gggacgtcct tctgctacgt cccttcggct ctcaatccag cggacctccc ttcccgaggc 5400
cttctgccgg ttctgcggcc tctcccgcgt cttcgctttc ggcctccgac gagtcggatc 5460
tccctttggg ccgcctcccc gcctgtttcg cctcggcgtc cggtccgtgt tgcttggtcg 5520
tcacctgtgc agaattgcga accatggatt ccaccgtgaa ctttgtctcc tggcatgcaa 5580
atcgtcaact tggcatgcca agaataattc ggatccaagc ttaggcctgc tcgctttctt 5640
gctgtcccat ttctattaaa ggttcctttg ttccctaagt ccaactacta aactggggga 5700
tattatgaag ggccttgagc atctggattc tgcctagcgc taagcttaac acgagccata 5760
gatagaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg 5820
taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg 5880
agaatagaga agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca 5940
ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagtt ggaacagcag 6000
aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa 6060
cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc 6120
cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg 6180
cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc 6240
tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg ttctcaataa 6300
accctcttgc agttgcatcc gactcgtggt ctcgctgttc cttgggaggg tctcctctga 6360
gtgattgact gcccacctcg ggggtctttc attctcgagc agcttggcgt aatcatggtc 6420
atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 6480
aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 6540
gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 6600
ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 6660
ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 6720
acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 6780
aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 6840
tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 6900
aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 6960
gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 7020
acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 7080
accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 7140
ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 7200
gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 7260
aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagt 7314
Claims (7)
1.一种TCR引物组,其特征在于,包括位点为L24-2和/或L24-49的引物对。
2.根据权利要求1所述的引物组,其特征在于,所述引物组依据HLAA24分型,EBV病毒抗原肽段IYVLVMLVL的TCR设计。
3.根据权利要求1所述的引物组,其特征在于,位点为L24-2的TCR引物对的序列如SEQID NO.1~2所示;
位点为L24-49的TCR引物对的序列如SEQ ID NO.3~4所示。
4.根据权利要求1所述的引物组,其特征在于,位点为L24-2的TCR引物对的序列还如SEQ ID NO.6~7所示;
位点为L24-49的TCR引物对的序列还如SEQ ID NO.8~9所示。
5.权利要求1~4任意一项所述的引物组在制备特异性检测HLAA24分型患者体内EBV病毒抗原肽段IYVLVMLVL的TCR产品中的应用。
6.根据权利要求5所述的应用,其特征在于,所述产品为试剂盒产品。
7.根据权利要求6所述的应用,其特征在于,所述试剂盒为荧光定量PCR试剂盒。
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