CN109706265B - 一种snp组合及其在桉树无性系鉴定中的应用 - Google Patents
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Abstract
本发明提供了一种SNP组合及其在桉树无性系鉴定中的应用,属于SNP标记技术领域。采用本发明提供的70个SNP标记组合极大提高对无性系检测准确度、缩短检测时间、提高检测水平,可有效应用于桉树无性系鉴定,快速甄别假冒和错乱的无性系,有效减少对桉树无性系鉴定的工作量,为桉树无性系推广、良种市场规范化提供强有力的技术检测,切实保障良种培育人和营林种植者的权益。
Description
技术领域
本发明涉及SNP标记技术领域,具体涉及一种SNP组合及其在桉树无性系鉴定中的应用。
背景技术
桉树作为南方重要的人工林树种,种质资源丰富,种源复杂,其种间杂交容易培育优良的杂交种,子代进行无性繁殖后可以培育大量无性系。目前我国桉树在历经多年的栽培和推广后,无性系关系混乱、良莠不齐等的问题日益严峻,不利于桉树的健康快速和有序发展。如何通过发展快速、高效的无性系鉴定方法来准确鉴定桉树无性系,达到桉树商品林良种市场规范化,切实维护育种者的权益显得极为重要。随着分子生物学的发展,应用分子标记技术对桉树进行分子水平上的鉴定,为桉树无性系的鉴定提供了新的手段。
目前无性系鉴定中常用的指纹图谱构建多数是基于微卫星(Simple SequenceRepeats,SSR)分子标记,但是,传统SSR在大规模基因分型时成本较高,人工耗时长,难以实现自动化、规模化自动检测,限制了其在林木分子育种中的大规模应用。作为第三代分子标记,SNP标记为克服这些障碍提供了一个很好的选择。相比于传统的SSR标记,SNP标记在基因组中更为丰富,为基因型筛选提供了大量的位点,具有许多优点,包括分布密度高、标记丰富、遗传性稳定性好等特点,更容易实现高通量和自动化检测,可以满足不同需求的各种基因分型系统。
随着高通量测序技术的发展,高通量的SNP检测技术在无性系鉴定方面具有广泛的应用前景,目前市面上已有较多的SNP基因分型高通量检测平台。巨桉的基因组测序已经完成,为了更好的促进桉树无性系的鉴定和育种应用,亟需一种通量高、重复性好的SNP应用于桉树无性系的快速、高效鉴定。
发明内容
本发明的目的在于提供了一种SNP组合及其在桉树无性系鉴定中的应用,本发明基于NCBI上的桉树测序数据开发SNP标记,利用Fluidigm的EP1高通量基因分析系统平台对主要桉树无性系品种进行基因分型,根据分型结果筛选出成功分型的SNP位点作为核心标记,并利用核心标记来构建无性系间的聚类图,可以更准确、更高效、高通量的鉴定桉树无性系并构建桉树无性系指纹图谱,从而达到快速甄别假冒和错乱的无性系、切实保障良种培育人和营林种植者的权益及提高桉树商品林发展良种化程度的目的。
本发明提供一种SNP组合,其特征在于,所述SNP包括:Chr01:1711907:Eucgr.A00084;Chr01:1855842:Eucgr.A00098;Chr01:1873036:Eucgr.A00102;Chr01:2969217:Eucgr.L00009;Chr01:5215832:Eucgr.H01866;Chr01:5216047:Eucgr.H01866;Chr01:35622575:Eucgr.A02019;Chr01:39099856:Eucgr.A02361;Chr01:39725463:Eucgr.A02437;Chr01:40501606:Eucgr.A02519;Chr01:41707457:Eucgr.A02641;Chr02:3701383:Eucgr.B00389;Chr02:7002640:Eucgr.B00669;Chr02:45050376:Eucgr.B02502;Chr02:51451355:Eucgr.B03117;Chr02:58699430:Eucgr.B03952;Chr03:83038236:Eucgr.C04060;Chr03:83337910:Eucgr.C04032;Chr04:134190:Eucgr.D00017;Chr04:31851675:Eucgr.D01833;Chr04:36183642:Eucgr.D02280;Chr04:36322007:Eucgr.D02302;Chr04:38246649:Eucgr.D02453;Chr04:38576688:Eucgr.D02475;Chr04:39978777:Eucgr.D02604;Chr04:40162642:Eucgr.D02626;Chr04:40431329:Eucgr.D02651;Chr05:1028097:Eucgr.E00114;Chr05:3394586:Eucgr.E00360;Chr05:6193636:Eucgr.E00656;Chr05:8497377:Eucgr.E00819;Chr05:16136396:Eucgr.E01477;Chr05:20437933:Eucgr.E01677;Chr05:46777811:Eucgr.E02861;Chr05:53151029:Eucgr.E03224;Chr05:54628397:Eucgr.E03312;Chr05:70874904:Eucgr.E04053;Chr05:75501905:Eucgr.E04303;Chr06:5029781:Eucgr.F00373;Chr06:37039557:Eucgr.F02490;Chr06:38459710:Eucgr.F02638;Chr06:38666572:Eucgr.F02649;Chr06:39211532:Eucgr.F02711;Chr06:42889891:Eucgr.F03107;Chr06:43784930:Eucgr.F03197;Chr06:56073835:Eucgr.F04359;Chr06:56566016:Eucgr.F04402;Chr07:9008359:Eucgr.G00611;Chr07:33966931:Eucgr.G01767;Chr07:37928892:Eucgr.G01960;Chr07:38192947:Eucgr.G01977;Chr07:40906121:Eucgr.G02115;Chr07:41507491:Eucgr.G02157;Chr07:41883821:Eucgr.G02184;Chr07:45261347:Eucgr.G02432;Chr07:45441706:Eucgr.G02457;Chr07:47129788:Eucgr.G02654;Chr08:2767734:Eucgr.H00455;Chr08:5621947:Eucgr.H00187;Chr08:61706046:Eucgr.H04541;Chr08:66469536:Eucgr.H04748;Chr08:66500358:Eucgr.H04753;Chr08:72367099:Eucgr.L00738;Chr10:675216:Eucgr.J00040;Chr10:1042073:Eucgr.J00068;Chr10:1140544:Eucgr.J00084;Chr10:1212759:Eucgr.J00090;Chr10:25955732:Eucgr.J02077;Chr11:16721043:Eucgr.K01335;Chr11:44430906:Eucgr.K03587。
本发明还提供了上述技术方案所述的SNP组合在桉树无性系鉴定中的应用。
优选的,所述应用包括:将上述技术方案所述SNP组合对桉树无性系的基因型进行分型,根据SNP分型结果利用软件R 3.4.3统计桉树无性系个体间的欧氏遗传距离,建立个体间聚类关系的遗传距离矩阵,按照个体间的遗传距离对桉树无性系进行聚类分析,通过分析个体间遗传距离的远近来鉴定桉树无性系,并构建桉树无性系间的聚类图,实现对桉树无性系的鉴定。
优选的,采用软件R3.4.3的R语言程序计算得出桉树无性系个体间的遗传矩阵;
所述R语言程序包括devtools、adegenet、hierfstat、pegas、poppr、ape和ade4。
优选的,所述桉树无性系的名称包括:Guanglin4、Guanglin5、Guang9、DH32-22、DH32-26、T13、EC4、DH32-32、668、EA14-12、EA14-01、EA14-06、DH32-27、DH101-2、DH184-1、DH32-21、DH33-9、k7、DH32-28、DH32-13、DH107-2、GL-WC1、LH88、LH1、Lei7、SH1、Lei11、Lei9、SH49、U7、A13、Q9、Z10-42、9224、U6、Eg6、GL9、Guangzhou1和D29。
本发明实施例的结果显示:采用本发明提供的70个SNP标记组合极大提高对无性系检测准确度、缩短检测时间、提高检测水平,可有效应用于桉树无性系鉴定,快速甄别假冒和错乱的无性系,有效减少对桉树无性系鉴定的工作量,为桉树无性系推广、良种市场规范化提供强有力的技术检测,切实保障良种培育人和营林种植者的权益。
附图说明
图1为部分桉树无性系基因分型结果;
图2为构建无性系间的聚类图。
具体实施方式
本发明提供了一种SNP组合,所述SNP包括:
Chr01:1711907:Eucgr.A00084;Chr01:1855842:Eucgr.A00098;Chr01:1873036:Eucgr.A00102;Chr01:2969217:Eucgr.L00009;Chr01:5215832:Eucgr.H01866;Chr01:5216047:Eucgr.H01866;Chr01:35622575:Eucgr.A02019;Chr01:39099856:Eucgr.A02361;Chr01:39725463:Eucgr.A02437;Chr01:40501606:Eucgr.A02519;Chr01:41707457:Eucgr.A02641;Chr02:3701383:Eucgr.B00389;Chr02:7002640:Eucgr.B00669;Chr02:45050376:Eucgr.B02502;Chr02:51451355:Eucgr.B03117;Chr02:58699430:Eucgr.B03952;Chr03:83038236:Eucgr.C04060;Chr03:83337910:Eucgr.C04032;Chr04:134190:Eucgr.D00017;Chr04:31851675:Eucgr.D01833;Chr04:36183642:Eucgr.D02280;Chr04:36322007:Eucgr.D02302;Chr04:38246649:Eucgr.D02453;Chr04:38576688:Eucgr.D02475;Chr04:39978777:Eucgr.D02604;Chr04:40162642:Eucgr.D02626;Chr04:40431329:Eucgr.D02651;Chr05:1028097:Eucgr.E00114;Chr05:3394586:Eucgr.E00360;Chr05:6193636:Eucgr.E00656;Chr05:8497377:Eucgr.E00819;Chr05:16136396:Eucgr.E01477;Chr05:20437933:Eucgr.E01677;Chr05:46777811:Eucgr.E02861;Chr05:53151029:Eucgr.E03224;Chr05:54628397:Eucgr.E03312;Chr05:70874904:Eucgr.E04053;Chr05:75501905:Eucgr.E04303;Chr06:5029781:Eucgr.F00373;Chr06:37039557:Eucgr.F02490;Chr06:38459710:Eucgr.F02638;Chr06:38666572:Eucgr.F02649;Chr06:39211532:Eucgr.F02711;Chr06:42889891:Eucgr.F03107;Chr06:43784930:Eucgr.F03197;Chr06:56073835:Eucgr.F04359;Chr06:56566016:Eucgr.F04402;Chr07:9008359:Eucgr.G00611;Chr07:33966931:Eucgr.G01767;Chr07:37928892:Eucgr.G01960;Chr07:38192947:Eucgr.G01977;Chr07:40906121:Eucgr.G02115;Chr07:41507491:Eucgr.G02157;Chr07:41883821:Eucgr.G02184;Chr07:45261347:Eucgr.G02432;Chr07:45441706:Eucgr.G02457;Chr07:47129788:Eucgr.G02654;Chr08:2767734:Eucgr.H00455;Chr08:5621947:Eucgr.H00187;Chr08:61706046:Eucgr.H04541;Chr08:66469536:Eucgr.H04748;Chr08:66500358:Eucgr.H04753;Chr08:72367099:Eucgr.L00738;Chr10:675216:Eucgr.J00040;Chr10:1042073:Eucgr.J00068;Chr10:1140544:Eucgr.J00084;Chr10:1212759:Eucgr.J00090;Chr10:25955732:Eucgr.J02077;Chr11:16721043:Eucgr.K01335;Chr11:44430906:Eucgr.K03587。
在本发明中,所述SNP位点名称命名格式为:染色体:SNP位置:基因名称。
本发明还提供了上述技术方案所述的SNP组合在桉树无性系鉴定中的应用。
在本发明中,所述应用优选包括:将上述技术方案所述SNP组合对桉树无性系的基因型进行分型,根据SNP分型结果利用软件R 3.4.3统计桉树无性系个体间的欧氏遗传距离,建立个体间聚类关系的遗传距离矩阵,按照个体间的遗传距离对桉树无性系进行聚类分析,通过分析个体间遗传距离的远近来鉴定桉树无性系,并构建桉树无性系间的聚类图,实现对桉树无性系的鉴定。
本发明优选采用软件R3.4.3的R语言程序计算得出桉树无性系个体间的遗传矩阵。在本发明中,所述R语言程序优选包括devtools、adegenet、hierfstat、pegas、poppr、ape和ade4。
下面结合具体实施例对本发明所述的一种SNP组合及其在桉树无性系鉴定中的应用做进一步详细的介绍,本发明的技术方案包括但不限于以下实施例。
实施例1
(1)SNP标记位点开发和探针设计:
利用NCBI上公布的尾叶桉和细叶桉测序数据(SRA编号:SRR1653562和SRR1653563),以已经发布的巨桉基因组序列(http://www.phytozome.net/eucalyptus.php)为参考基因组,使用GATK v3.5来识别SNPs,使用vcftools v4.0来筛选过滤SNP位点,查找位于功能基因区域的多态SNP位点,从中筛选出70个在染色体上均匀分布的SNP位点。根据在巨桉参考基因组上的位置,提取70个SNP标记的侧翼序列位点上下游各100bp,定制富鲁达SNP芯片及交由Fluidigm公司合成用于检测70个SNP位点的探针(如表1和2所示)。
表1 70个SNP探针序列
表2 70个SNP探针序列
(2)桉树无性系DNA的提取:
收集桉树无性系叶片(表3),利用CTAB法提取桉树基因组DNA,置于冰箱冷冻后备用。利用CTAB法提取桉树基因组DNA,置于-80℃冰箱保存备用。
表3 39份桉树无性系材料
(3)DNA样品质量检测:
用2%的琼脂糖凝胶电泳检测所有无性系样品DNA的完整性,用凝胶成像系统判断电泳结果,确保基因组DNA完整性好,用Nanodrop 2000检测基因组中蛋白质和有机物质污染水平,DNA的OD260/OD280比值应在1.8~2.0之间,OD260/OD230比值应在1.8~2.2之间,统一将样品DNA的浓度稀释到工作浓度60ng/μl。
(3)SNP标记位点的基因分型试验:
利用Fluidigm EP1高通量基因分析系统平台,采用Fluidigm 96.96(IntegratedFluidic Circuit,IFC)芯片。首先参照Fluidigm实验方案STA操作流程(SNP分型说明书,http://cn.fluidigm.com/),将上述得到的基因组DNA按照操作说明进行目标特异性靶扩增(Specific Target Amplification,STA)处理得到扩增产物,之后将扩增产物统一用DNASuspension Buffer稀释至20×。
将稀释后的扩增产物参照Fluidigm基因分型试验方案说明操作(SNP分型说明书,http://cn.fluidigm.com/)进行基因分型实验。
(4)数据分析:
使用Fluidigm数据处理软件扫描结果,采集图像和数据,使用FluidigmSNP分型软件分析所有无性系样品的基因型。按照质控阈值SNP calling rate大于85%,对缺失和低质量分型数据进行人工复检。统计39份无性系样品的基因分型结果,筛选SNP callingrate值均大于85%及样品缺失率小于5%的SNP标记位点,部分结果桉树无性系基因分型结果如图1所示。统计分析结果,筛选出70个SNP位点作为核心SNP标记:Chr01:1711907:Eucgr.A00084;Chr01:1855842:Eucgr.A00098;Chr01:1873036:Eucgr.A00102;Chr01:2969217:Eucgr.L00009;Chr01:5215832:Eucgr.H01866;Chr01:5216047:Eucgr.H01866;Chr01:35622575:Eucgr.A02019;Chr01:39099856:Eucgr.A02361;Chr01:39725463:Eucgr.A02437;Chr01:40501606:Eucgr.A02519;Chr01:41707457:Eucgr.A02641;Chr02:3701383:Eucgr.B00389;Chr02:7002640:Eucgr.B00669;Chr02:45050376:Eucgr.B02502;Chr02:51451355:Eucgr.B03117;Chr02:58699430:Eucgr.B03952;Chr03:83038236:Eucgr.C04060;Chr03:83337910:Eucgr.C04032;Chr04:134190:Eucgr.D00017;Chr04:31851675:Eucgr.D01833;Chr04:36183642:Eucgr.D02280;Chr04:36322007:Eucgr.D02302;Chr04:38246649:Eucgr.D02453;Chr04:38576688:Eucgr.D02475;Chr04:39978777:Eucgr.D02604;Chr04:40162642:Eucgr.D02626;Chr04:40431329:Eucgr.D02651;Chr05:1028097:Eucgr.E00114;Chr05:3394586:Eucgr.E00360;Chr05:6193636:Eucgr.E00656;Chr05:8497377:Eucgr.E00819;Chr05:16136396:Eucgr.E01477;Chr05:20437933:Eucgr.E01677;Chr05:46777811:Eucgr.E02861;Chr05:53151029:Eucgr.E03224;Chr05:54628397:Eucgr.E03312;Chr05:70874904:Eucgr.E04053;Chr05:75501905:Eucgr.E04303;Chr06:5029781:Eucgr.F00373;Chr06:37039557:Eucgr.F02490;Chr06:38459710:Eucgr.F02638;Chr06:38666572:Eucgr.F02649;Chr06:39211532:Eucgr.F02711;Chr06:42889891:Eucgr.F03107;Chr06:43784930:Eucgr.F03197;Chr06:56073835:Eucgr.F04359;Chr06:56566016:Eucgr.F04402;Chr07:9008359:Eucgr.G00611;Chr07:33966931:Eucgr.G01767;Chr07:37928892:Eucgr.G01960;Chr07:38192947:Eucgr.G01977;Chr07:40906121:Eucgr.G02115;Chr07:41507491:Eucgr.G02157;Chr07:41883821:Eucgr.G02184;Chr07:45261347:Eucgr.G02432;Chr07:45441706:Eucgr.G02457;Chr07:47129788:Eucgr.G02654;Chr08:2767734:Eucgr.H00455;Chr08:5621947:Eucgr.H00187;Chr08:61706046:Eucgr.H04541;Chr08:66469536:Eucgr.H04748;Chr08:66500358:Eucgr.H04753;Chr08:72367099:Eucgr.L00738;Chr10:675216:Eucgr.J00040;Chr10:1042073:Eucgr.J00068;Chr10:1140544:Eucgr.J00084;Chr10:1212759:Eucgr.J00090;Chr10:25955732:Eucgr.J02077;Chr11:16721043:Eucgr.K01335;Chr11:44430906:Eucgr.K03587。
(5)无性系指纹图谱构建:
基于核心70个SNP标记组合对39个无性系的基因分型结果,利用软件R 3.4.3,采用R语言程序包devtools、adegenet、hierfstat、pegas、poppr、ape、ade4来计算无性系个体间的遗传矩阵,对39个无性系进行聚类分析,鉴定39个无性系的真实性和一致性,构建39株无性系间的聚类图如图2所示,聚类结果表明,在遗传距离为9.89时,可以将39个无性系划分为四大类,根据聚类图从上往下划分可将无性系Guanglin4、Guanglin5、DH32-22、T13、Guang9、EC4、GL9、DH32-26、DH33-9、DH32-21、U7、DH32-13、DH32-27、DH32-22、DH32-28、Q9、Z10-42、Eg6、A13、Lei9归为第一类;无性系Lei7、LH88、EA14-12归为第二类;无性系9224、LH1、Guangzhou1、DH107-2、DH184-1、Lei11、SH49、U6、GL-WC1可归为第三类;无性系D29、SH1、EA14-01、668、k7归为第四类。其中39个无性系中,无性系LH1和Guangzhou1的遗传距离最小,说明这两个无性系之间的亲缘关系最近;Lei7的遗传距离最大,说明无性系Lei7为尾赤桉与其他无性系直接的亲缘关系较远。在第一类无性系中,各无性系间的遗传距离都比较接近,根据表2无性系的名称,推测出第一类的无性系很有可能为尾巨桉,第二类无性系中,Lei7为尾赤桉,其中EA14-12与LH88的遗传距离相近,其中LH88为尾叶桉,推测EA14-12应为尾叶桉或为与尾叶桉亲缘关系相近的其他桉属树种。综上,利用70个SNP标记组合可对39株无性系进行有效鉴定,成功构建出39个无性系间的遗传聚类图,从一定程度上反映了各个无性系之间的遗传差异程度,并鉴定出T13、Guang9、EC4、GL9为同一个无性系及DH32-26和DH33-9也为同一无性系,可为今后桉树无性系的快速鉴定提供有效的参考。
由以上实施例可以得出,采用本发明方法构建了39个国内常用桉树无性系指纹图谱,并成功的对上述无性系进行了鉴定;基于70个SNP标记组合基础上开发的通用型中通量Fluidigm SNP芯片将极大提高对无性系检测准确度、缩短检测时间、提高检测水平,可有效应用于桉树无性系鉴定,快速甄别假冒和错乱的无性系,有效减少对桉树无性系鉴定的工作量,为桉树无性系推广、良种市场规范化提供强有力的技术检测,切实保障良种培育人和营林种植者的权益。
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> 中国林业科学研究院热带林业研究所
<120> 一种SNP组合及其在桉树无性系鉴定中的应用
<160> 280
<170> SIPOSequenceListing 1.0
<210> 1
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
tgctagtgta agaccatatg aatgca 26
<210> 2
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
actgacccga tcatgatcat cg 22
<210> 3
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
agcagcagca gttacagcat 20
<210> 4
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
ggcattcaaa aggaggtttt gcata 25
<210> 5
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
catcggaacc ccatttcaag t 21
<210> 6
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
agtcttttcg aattatggtt atggtgc 27
<210> 7
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
gagctaggaa ttctgagtgc tcat 24
<210> 8
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
cccaatccaa gtcattgtgg ac 22
<210> 9
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
tcaagttaga ccgtatcatt atgatagcc 29
<210> 10
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
gctgtggaag aaaccgattg tc 22
<210> 11
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
gcccatttca atatgtgcca ca 22
<210> 12
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
cccaatagac atacagtttg gtaggc 26
<210> 13
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
cacaagtagc aatttcaaga acattatcgt 30
<210> 14
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
tcgaaacact ggccacga 18
<210> 15
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
ccgcaagctt tttcctcaag c 21
<210> 16
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
aatatcacaa ctattatgga gggtgactc 29
<210> 17
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
tcgcgaggaa gtacggtttt 20
<210> 18
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 18
ctccaattgc aggagcagag a 21
<210> 19
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 19
cttcaagatc gagtcttgga aacg 24
<210> 20
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 20
accgcccttg gtatgtgg 18
<210> 21
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
tcaggcacaa ccagacagaa 20
<210> 22
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 22
gttggtttct tatggcaagc aaga 24
<210> 23
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 23
attttcaaaa ggagagcaaa actcgaa 27
<210> 24
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 24
ggaaaaaaga gatttcgcgg aga 23
<210> 25
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 25
cacctgctac agctctgtca at 22
<210> 26
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 26
gagaagtatc attgaaagaa cataaaacag agaa 34
<210> 27
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 27
ctggtgttgg aacaacactt ctc 23
<210> 28
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 28
cttgatttca accagggtgg ttaaag 26
<210> 29
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 29
gccaatgcct tgtttggcc 19
<210> 30
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 30
tggccttctc acaggttctt t 21
<210> 31
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 31
cagtcatcca ttatcccatg agtttg 26
<210> 32
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 32
ggcatgaaac atccaaaagc atc 23
<210> 33
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 33
ggaaaaagac agtagctcaa gtgac 25
<210> 34
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 34
ttactctaaa atcatcacaa gtgaagaaaa taa 33
<210> 35
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 35
tgcatcacaa gcaaagctca taatc 25
<210> 36
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 36
ccttctatct ccgatgcgtc tc 22
<210> 37
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 37
ggatggaacc tcttcctctc aa 22
<210> 38
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 38
cccgagcgat tcctccg 17
<210> 39
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 39
ggaatttgac ctgattcatg cttttg 26
<210> 40
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 40
ctacataaga agcagaaatg ctaagaaca 29
<210> 41
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 41
tctgtttggt ctgttggtct gtt 23
<210> 42
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 42
tcctcgcgag attttccttc ac 22
<210> 43
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 43
ctaaggatcc cacaagaaac aaatcaa 27
<210> 44
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 44
tcacattgat aaatcttttt ggctatggc 29
<210> 45
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 45
ccagatctca tcccatcttt actgac 26
<210> 46
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 46
cctattgact gtgaatttgt gtcttttctc 30
<210> 47
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 47
gttctcttct agtagaggtg caacg 25
<210> 48
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 48
cacatgcatc gacaaaatcc tcg 23
<210> 49
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 49
ttctgctcgg aggtgtgaac 20
<210> 50
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 50
tcctcccatg gaatggcg 18
<210> 51
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 51
tggctgaaaa actaaaaggc cg 22
<210> 52
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 52
acttgaaatc gacaatatat ggcatttaag tag 33
<210> 53
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 53
tggtggtcga gcccaac 17
<210> 54
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 54
cttcttttct gatgtgttgc tagaaact 28
<210> 55
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 55
cgcggtgaat tcaacacacg 20
<210> 56
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 56
gtccttttac ctgtatgcat acacatttaa t 31
<210> 57
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 57
tgtgtcttta tgtctaatgt agcatgtaga t 31
<210> 58
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 58
gcttcttagt gatattaact ggatgcg 27
<210> 59
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 59
caattttgtt gctcaattta tttgtttgaa agag 34
<210> 60
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 60
acccaagttt ggaccgct 18
<210> 61
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 61
ccttgggtcg cagtgtgta 19
<210> 62
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 62
ccttcttttt ttcagggttc tggt 24
<210> 63
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 63
acttcttcga cctccccca 19
<210> 64
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 64
acctgagctt ttgagggttg ttta 24
<210> 65
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 65
gcgcacgagt ccatgatc 18
<210> 66
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 66
gtcatttact ttagctcttc tatgagctct a 31
<210> 67
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 67
agtcctcttc ctcggtctca 20
<210> 68
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 68
gccgatgcaa attggaagct c 21
<210> 69
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 69
tcatcatcta cgcaagggac g 21
<210> 70
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 70
agaagaagag acttgaaact ttccct 26
<210> 71
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 71
tgctagtgta agaccatatg aatgcg 26
<210> 72
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 72
actgacccga tcatgatcat cc 22
<210> 73
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 73
gcagcagcag ttacagcac 19
<210> 74
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 74
gcattcaaaa ggaggttttg catg 24
<210> 75
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 75
catcggaacc ccatttcaag g 21
<210> 76
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 76
cagtcttttc gaattatggt tatggtgt 28
<210> 77
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 77
agctaggaat tctgagtgct cag 23
<210> 78
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 78
acccaatcca agtcattgtg gat 23
<210> 79
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 79
tttcaagtta gaccgtatca ttatgatagc t 31
<210> 80
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 80
gctgtggaag aaaccgattg tt 22
<210> 81
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 81
gcccatttca atatgtgcca cg 22
<210> 82
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 82
cccaatagac atacagtttg gtaggt 26
<210> 83
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 83
acaagtagca atttcaagaa cattatcgg 29
<210> 84
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 84
tcgaaacact ggccacgg 18
<210> 85
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 85
ccgcaagctt tttcctcaag t 21
<210> 86
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 86
aatatcacaa ctattatgga gggtgactt 29
<210> 87
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 87
tcgcgaggaa gtacggtttc 20
<210> 88
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 88
tccaattgca ggagcagagg 20
<210> 89
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 89
ccttcaagat cgagtcttgg aaaca 25
<210> 90
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 90
accgcccttg gtatgtgt 18
<210> 91
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 91
tcaggcacaa ccagacagac 20
<210> 92
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 92
ttggtttctt atggcaagca agg 23
<210> 93
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 93
ttttcaaaag gagagcaaaa ctcgag 26
<210> 94
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 94
ggaaaaaaga gatttcgcgg agc 23
<210> 95
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 95
acctgctaca gctctgtcaa g 21
<210> 96
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 96
gagaagtatc attgaaagaa cataaaacag agac 34
<210> 97
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 97
ctggtgttgg aacaacactt ctg 23
<210> 98
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 98
cttgatttca accagggtgg ttaaaa 26
<210> 99
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 99
gccaatgcct tgtttggct 19
<210> 100
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 100
tggccttctc acaggttctt g 21
<210> 101
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 101
tcagtcatcc attatcccat gagtttt 27
<210> 102
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 102
gggcatgaaa catccaaaag catt 24
<210> 103
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 103
ggaaaaagac agtagctcaa gtgag 25
<210> 104
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 104
ttactctaaa atcatcacaa gtgaagaaaa tat 33
<210> 105
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 105
aatgcatcac aagcaaagct cataata 27
<210> 106
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 106
ccttctatct ccgatgcgtc tt 22
<210> 107
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 107
ggatggaacc tcttcctctc ag 22
<210> 108
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 108
cccgagcgat tcctcct 17
<210> 109
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 109
gggaatttga cctgattcat gctttta 27
<210> 110
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 110
ctacataaga agcagaaatg ctaagaacg 29
<210> 111
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 111
tctgtttggt ctgttggtct gtc 23
<210> 112
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 112
tcctcgcgag attttccttc aa 22
<210> 113
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 113
ctaaggatcc cacaagaaac aaatcag 27
<210> 114
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 114
tcacattgat aaatcttttt ggctatggg 29
<210> 115
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 115
ccagatctca tcccatcttt actgat 26
<210> 116
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 116
cctattgact gtgaatttgt gtcttttctg 30
<210> 117
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 117
gttctcttct agtagaggtg caaca 25
<210> 118
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 118
cacatgcatc gacaaaatcc tca 23
<210> 119
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 119
attctgctcg gaggtgtgaa t 21
<210> 120
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 120
tttcctccca tggaatggca 20
<210> 121
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 121
ctggctgaaa aactaaaagg cca 23
<210> 122
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 122
acttgaaatc gacaatatat ggcatttaag tac 33
<210> 123
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 123
ctggtggtcg agcccaat 18
<210> 124
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 124
ttcttttctg atgtgttgct agaaacc 27
<210> 125
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 125
cgcggtgaat tcaacacaca 20
<210> 126
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 126
gtccttttac ctgtatgcat acacatttaa a 31
<210> 127
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 127
tgtgtcttta tgtctaatgt agcatgtaga c 31
<210> 128
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 128
ggcttcttag tgatattaac tggatgca 28
<210> 129
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 129
tcaattttgt tgctcaattt atttgtttga aagaa 35
<210> 130
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 130
cccaagtttg gaccgcc 17
<210> 131
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 131
cttgggtcgc agtgtgtg 18
<210> 132
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 132
ccttcttttt ttcagggttc tgga 24
<210> 133
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 133
cttcttcgac ctcccccc 18
<210> 134
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 134
cctgagcttt tgagggttgt ttt 23
<210> 135
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 135
gcgcacgagt ccatgatg 18
<210> 136
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 136
tcatttactt tagctcttct atgagctctg 30
<210> 137
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 137
agtcctcttc ctcggtctcg 20
<210> 138
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 138
gccgatgcaa attggaagct g 21
<210> 139
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 139
ctcatcatct acgcaaggga ca 22
<210> 140
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 140
agaagaagag acttgaaact ttcccc 26
<210> 141
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 141
cgcatatggc tttctgagct tacatt 26
<210> 142
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 142
cccgccgcca gaagaag 17
<210> 143
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 143
ccagctggtt gctgctgt 18
<210> 144
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 144
ccctccaaag cttgccattg t 21
<210> 145
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 145
gcagcttcag cttatccttg cc 22
<210> 146
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 146
gccaagattc ataataaagc cacc 24
<210> 147
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 147
ttccccgtcc atctttggtc t 21
<210> 148
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 148
agcgaacgct cttaacaccc t 21
<210> 149
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 149
ctggcctaat atgttatcca agagatctaa gaa 33
<210> 150
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 150
cctgcccatt tcaatatgtg cca 23
<210> 151
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 151
tggaagaaac cgattgttgc t 21
<210> 152
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 152
tgcttgagac cagggaatga ctc 23
<210> 153
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 153
agcagtggac gttttggtcc ta 22
<210> 154
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 154
agagcgactt cctcgcctc 19
<210> 155
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 155
acgtgtctcg gagaattttg gct 23
<210> 156
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 156
cagctttatg aattctgttc ccaacact 28
<210> 157
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 157
gggttctcgg gcatttccc 19
<210> 158
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 158
agggcttcct tgagcgaaga a 21
<210> 159
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 159
gcgaacaaca ggcgaccc 18
<210> 160
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 160
gctttcttat gctagaatct gaaacacatt ga 32
<210> 161
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 161
ccaatcatgt gagatcttca aaaagggt 28
<210> 162
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 162
gaatcctcag ttcccctttg atag 24
<210> 163
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 163
agcagtttga gtgtccaggc a 21
<210> 164
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 164
ctcaacggcg aaacaccgat 20
<210> 165
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 165
gactcaggaa ataaagactt ccttccct 28
<210> 166
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 166
tctggataca atgaatgtag agcacttcc 29
<210> 167
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 167
tgcctcagga agctcactgt 20
<210> 168
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 168
gcaccaggta ccttcaatgg c 21
<210> 169
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 169
gcatgctcct tgtagcactc c 21
<210> 170
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 170
caacctcagt gtgcgacttg t 21
<210> 171
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 171
tggttcaatt tgattctact tgaaaccaaa ttgt 34
<210> 172
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 172
ccaaaggttt taattccagc tataaaatta ccca 34
<210> 173
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 173
ctggtgtgct catctctcct gt 22
<210> 174
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 174
tcctgttgct tggattgttt gtgac 25
<210> 175
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 175
gaatcatttg acactgcagc tcct 24
<210> 176
<211> 16
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 176
cgacggaggc ccggaa 16
<210> 177
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 177
gctggaggta catctacctt ctgt 24
<210> 178
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 178
gttcttggcc ccgatggtg 19
<210> 179
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 179
gcaacaccaa aacttcctga tacct 25
<210> 180
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 180
accataccgt ttcctctgcg t 21
<210> 181
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 181
actgtcgtcc tttctgccca 20
<210> 182
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 182
cccaattccc ttcattgcgc t 21
<210> 183
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 183
tgttaagagt atcatttctt tcggttatcc agt 33
<210> 184
<211> 40
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 184
gaaagattag atcactgtat agaaaacaga tacttaccaa 40
<210> 185
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 185
cttgtcataa gtcatagcat catttaatgc ca 32
<210> 186
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 186
cgtctcactt catctgcaag atcct 25
<210> 187
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 187
aacctagaga aagggatgaa ttgtt 25
<210> 188
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 188
cgctgatcgt ctgcaggtc 19
<210> 189
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 189
cccagctgca ccggatg 17
<210> 190
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 190
catccccgtg atcatagcgg a 21
<210> 191
<211> 40
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 191
gtagatattg gttatctcag ctgtttattt atatagcttt 40
<210> 192
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 192
agtgaagctt aaataatagc tacaatgtct gaaca 35
<210> 193
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 193
gccgaacaag cggttcca 18
<210> 194
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 194
ggaatggaac tgttcaaaat ctgagataaa tcc 33
<210> 195
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 195
gcggcttcag ttcgcct 17
<210> 196
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 196
agcagattat gagcttggct tatttgc 27
<210> 197
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 197
tcactgatta caattgcttt gatgcaca 28
<210> 198
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 198
ccggtctccg gtgaagtca 19
<210> 199
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 199
gcaaatagca tgatatatga acttgaaacc tga 33
<210> 200
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 200
cggcgactac tacaacgcct 20
<210> 201
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 201
gggtcccgga ctgtgaca 18
<210> 202
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 202
cccagcacca cccacaag 18
<210> 203
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 203
ggcttggcgt aggcctc 17
<210> 204
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 204
ctggttggcc aaaaccctca t 21
<210> 205
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 205
caaccgagat catcacagtc cct 23
<210> 206
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 206
aaggccacct agattttctg tcca 24
<210> 207
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 207
ctgcagagac tgaagcttcc tct 23
<210> 208
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 208
ctagcaacac tagggttcat cg 22
<210> 209
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 209
gaagacatgc acaagaggcg a 21
<210> 210
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 210
cccagtgttc ctaagcaacc g 21
<210> 211
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 211
aagcattaga tttactatca aaagttcagt 30
<210> 212
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 212
ctcttgagct tccacttaaa tcca 24
<210> 213
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 213
gcatcaactg catcaccaac tt 22
<210> 214
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 214
gcttgagtcc cgcccatta 19
<210> 215
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 215
ccgatcatgg gaagatggc 19
<210> 216
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 216
tgcttgtatg gttattgaat atggaca 27
<210> 217
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 217
agtcgagcat tgggatattg ga 22
<210> 218
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 218
ccacctgctc actgatcaaa a 21
<210> 219
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 219
cctctctgtc caaattattt agttaaaaat gt 32
<210> 220
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 220
attgtcatta ggatcctttc acagc 25
<210> 221
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 221
caaccggatc atcagcacc 19
<210> 222
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 222
agctttccct cagtaattaa accc 24
<210> 223
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 223
aacgtattcg gtgtggaaaa ctc 23
<210> 224
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 224
ggcccgattt ttccggt 17
<210> 225
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 225
gtttactaga tagttgtgcc gca 23
<210> 226
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 226
caatatcaca actattatgg agggtgac 28
<210> 227
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 227
cttcatagac tcctccattg aggt 24
<210> 228
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 228
agagttctgg aggttcagct 20
<210> 229
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 229
cccagttttc cgaagtcctt tt 22
<210> 230
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 230
gcaaggaggt ctctgaagaa ca 22
<210> 231
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 231
tctgagtcag gcacaacca 19
<210> 232
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 232
gttccttcca tcagttttct ggtat 25
<210> 233
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 233
gcttagcaac ttatctaaag aaagacga 28
<210> 234
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 234
ggtgctcgag tctttcgg 18
<210> 235
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 235
catttgaagc agttaaattt ccacct 26
<210> 236
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 236
gggattgtaa tttcgagctt ccg 23
<210> 237
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 237
gtgcatgaat tctggatacc cac 23
<210> 238
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 238
aggtaactgg ttagaaggaa gctaa 25
<210> 239
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 239
gagttcaagc ttccttacca gaa 23
<210> 240
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 240
gtacctcgag ccatcgct 18
<210> 241
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 241
cggagtattg gcaggtattc a 21
<210> 242
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 242
tgagcagata attctttgac aaggg 25
<210> 243
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 243
caaaccagca agagggagaa aa 22
<210> 244
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 244
ttactctaaa atcatcacaa gtgaagaaa 29
<210> 245
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 245
atgagcaatt tgcaatacca atgc 24
<210> 246
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 246
cttcctccaa tctcacaatg tgg 23
<210> 247
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 247
tccctgggca gttctacg 18
<210> 248
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 248
acctcccaag acgagcac 18
<210> 249
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 249
agggtgtagt aattaagtct cgca 24
<210> 250
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 250
ccacttttat tttgccttaa ggtcac 26
<210> 251
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 251
cccagaggtc agaggcg 17
<210> 252
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 252
gcttccgtcg aatctcaatc t 21
<210> 253
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 253
tgaccgacca tgaaattctt caaat 25
<210> 254
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 254
ccttctattc gctgacacaa agc 23
<210> 255
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 255
ggactcattt cagggaaatc ca 22
<210> 256
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 256
ctgggctcct attgactgtg 20
<210> 257
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 257
gggtccacta gcatatttga atcat 25
<210> 258
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 258
gcatgtgtga ccccgac 17
<210> 259
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 259
cccttttgtt ccttcgggat 20
<210> 260
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 260
gcgacgaaaa ctccgatttc 20
<210> 261
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 261
tcccatgttt ccaactattc aactt 25
<210> 262
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 262
gccatgaaag gaaaaaagga aaaca 25
<210> 263
<211> 16
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 263
cgcgcataac gccgag 16
<210> 264
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 264
tgcactgaca tttatagcac ttcc 24
<210> 265
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 265
tcgcgtcgaa cgagagag 18
<210> 266
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 266
acatcaactt catcatcttc atgaaca 27
<210> 267
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 267
ttgtgtcttt atgtctaatg tagcatgt 28
<210> 268
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 268
gtgggccaat tcacaggg 18
<210> 269
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 269
gccttcagcg tataaattac gcttc 25
<210> 270
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 270
ggtttgattc gggactgacc 20
<210> 271
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 271
tcaagggctt gagaagcatt aag 23
<210> 272
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 272
cgctttttca atttttggga ttttcc 26
<210> 273
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 273
agctcatctc ccgcctg 17
<210> 274
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 274
tttaatggag aacattgacc tgagc 25
<210> 275
<211> 16
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 275
aggacgacca cgacgc 16
<210> 276
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 276
gcacttaagt cctaagactt gtgat 25
<210> 277
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 277
ccacagtgat cgtgaaggtg 20
<210> 278
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 278
tcttccagcg aaagaaattg actt 24
<210> 279
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 279
accaaccact gacccaga 18
<210> 280
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 280
agacacattg gattttcctt tacttga 27
Claims (3)
1.SNP探针在桉树无性系鉴定中的应用,其特征在于,所述SNP探针的核苷酸序列如SEQID No.1~SEQ ID No.280所示;
所述桉树无性系包括:Guanglin4、Guanglin5、Guang9、DH32-22、DH32-26、T13、EC4、DH32-32、668、EA14-12、EA14-01、EA14-06、DH32-27、DH101-2、DH184-1、DH32-21、DH33-9、k7、DH32-28、DH32-13、DH107-2、GL-WC1、LH88、LH1、Lei7、SH1、Lei11、Lei9、SH49、U7、A13、Q9、Z10-42、9224、U6、Eg6、GL9、Guangzhou1和D29。
2.根据权利要求1所述的应用,其特征在于,所述应用包括:利用权利要求1所述的SNP探针对桉树无性系的基因型进行分型,根据SNP分型结果利用软件R 3.4.3统计桉树无性系个体间的欧氏遗传距离,建立个体间聚类关系的遗传距离矩阵,按照个体间的遗传距离对桉树无性系进行聚类分析,通过分析个体间遗传距离的远近来鉴定桉树无性系,并构建桉树无性系间的聚类图,实现对桉树无性系的鉴定。
3.根据权利要求2所述的应用,其特征在于,采用软件R3.4.3的R语言程序计算得出桉树无性系个体间的遗传矩阵;
所述R语言程序包括devtools、adegenet、hierfstat、pegas、poppr、ape和ade4。
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