CN112708628A - 玉米百粒重主效QTL位点qKW4a及其候选基因和应用 - Google Patents

玉米百粒重主效QTL位点qKW4a及其候选基因和应用 Download PDF

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CN112708628A
CN112708628A CN202110068470.1A CN202110068470A CN112708628A CN 112708628 A CN112708628 A CN 112708628A CN 202110068470 A CN202110068470 A CN 202110068470A CN 112708628 A CN112708628 A CN 112708628A
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陶勇生
段会军
张大效
马小林
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Abstract

本发明公开了玉米百粒重主效QTL位点qKW4a及其候选基因和应用，属于作物遗传育种领域。本发明以SL41和Ye478为亲本构建遗传分离群体，通过图位克隆，将玉米百粒重主效QTL位点qKW4a精细定位于B73参考基因组chr4标记kw4–kw10之间，并且基于精细定位区段内基因表达模式、双亲间(SL41和Ye478)基因组和cDNA序列差异，以及籽粒淀粉形成过程中的酶活性和细胞学特征等确认了qKW4a的候选基因ZmSSV。本发明经图位克隆方法分离和克隆的候选基因ZmSSV与玉米KW(kernel weight)相关，将有效用于玉米KW的分子育种和遗传改良。

Description

玉米百粒重主效QTL位点qKW4a及其候选基因和应用
技术领域
本发明涉及作物遗传育种领域，具体而言，涉及玉米百粒重主效QTL位点qKW4a及其候选基因和应用。
背景技术
玉米百粒重(100-kernel weight,100-KW；或kernel weight,KW)是单穗产量的主要决定因子。所以，粒重QTL定位(或称图位克隆)和其候选基因的克隆将有助于我国粮食产量提高和粮食安全。玉米KW属多基因控制的数量性状，而且图位克隆是公认的解析数量性状遗传机制和候选基因分离的有效手段。并且已有研究表明玉米KW可由少数主效QTL控制。
玉米KW通常为籽粒干物质积累，而且以淀粉为主。胚乳细胞分裂期和籽粒充实期基因分离和克隆为解析淀粉含量、组成和结构，以及作物KW形成奠定了基础。如，细胞分裂素(cytokinins,CKs)及代谢途径相关基因，以及淀粉生物合成的关键酶的基因(包括蔗糖合成酶(SS)、ADP-葡萄糖焦磷酸化酶(AGPase)、Q酶(BE)、可溶性淀粉合成酶(SSS)、淀粉脱支酶(DBE)和糖苷水解酶(GH))。尽管这些基因分离和克隆为KW形成的分子调控奠定了基础，但对KW形成的分子机制的了解还远不够充分。
玉米KW属多基因控制的数量性状，而QTL(quantitative trait loci)定位(或称图位克隆)是公认的分离和确认数量性状候选基因的有效手段。现有技术中尚未有基于图位克隆实现KW主效QTL位点(qKW4a)的精细定位和候选基因确认的相关报道道。
发明内容
本发明的目的是提供一种玉米百粒重主效QTL位点qKW4a及其候选基因的分离和克隆及应用，以解决上述现有技术存在的问题，利用图位克隆将玉米百粒重主效QTL位点qKW4a精细定位于B73参考基因组chr4标记kw4–kw10之间，并得到其候选基因ZmSSV，为后续玉米KW的分子育种和遗传改良提供有效的工具。
为实现上述目的，本发明提供了如下方案：
本发明提供一种调控玉米百粒重主效QTL定位，所述的玉米百粒重主效QTL为qKW4a，位于B73参考基因组4号染色体上，分子标记kw4和kw10之间。
本发明提供所述的调控玉米百粒重主效QTL的候选基因，所述候选基因为ZmSSV，其基因序列为如下序列之一：
1)SEQ ID NO.1所示的序列；
2)在SEQ ID NO.1所示序列基础上发生一个或多个碱基的取代、缺失或替换且与SEQ ID NO.1所示的序列具有95％以上同一性的序列。
本发明还提供所述的调控玉米百粒重主效QTL或所述的候选基因在玉米百粒重性状的遗传改良和分子育种中的应用。
本发明公开了以下技术效果：
本发明以玉米染色体片段代换系纯合系SL41组配衍生群体用于KW的QTL(qKW4a)的精细定位，利用图位克隆将玉米百粒重主效QTL位点qKW4a精细定位于B73参考基因组chr4标记kw4–kw10之间，同时基于精细定位区段内基因表达模式、双亲间(SL41和Ye478)基因组和cDNA序列差异，以及籽粒淀粉形成过程中的酶活性确认了本专利经图位克隆方法分离和克隆的qKW4a的候选基因ZmSSV与玉米KW相关，将有效用于玉米KW的分子育种和遗传改良。
附图说明
图1为玉米籽粒qKW4a的精细定位和候选基因的分离克隆及其关联分析，图中A为qKW4a的初定位示意图；B为利用BC1群体对qKW4a的重定位示意图；C为SL41和Y478间差异的10个SSR标记；D为Zm00001d051976和Zm00001d051977的基因序列比对示意图；E为2个关联群体Zm00001d051976区段的关联分析图。
图2为双亲间候选基因Zm00001d051976的主要差异序列，A、B、C中上侧和下侧序列分别为SL41和Y478的基因组扩增序列；*表示序列相同碱基；深色背景部分为主要差异序列部分；图中A为基因序列中第6530-6536位碱基；B为第8559-8571位碱基；C为第13449-13455位碱基。
具体实施方式
本发明所使用材料，如无特殊说明，均可由商业途径获得。本发明所使用的实验方法，如无特殊说明，均为本领域常规方法。
实施例1
1、玉米KW主效QTL位点qKW4a精细定位
本发明基于玉米KW的QTL初定位结果实施(基于掖478导入系的玉米百粒重QTL鉴定，詹晶晶等,2015)，即控制KW的位点qKW4a定位于玉米4.07bin的bnlg1784-umc1194间的主效QTL位点，其表型贡献率17.81％(图1A)。
本发明在前期研究(基于掖478导入系的玉米百粒重QTL鉴定，詹晶晶等,2015)的基础上，利用210个BC1群体(由Y478和SL41杂交衍生构建，其中Y478为我国玉米优良自交系掖478，SL41是以Y478为遗传背景导入QB80的染色体单片段代换系)，进行每一个单株已开发标记检测，进而获得基因型，同时田间种植鉴定每个单株KW表型，将qKW4a重定位于umc2038-umc1194的B73参考基因组5.20Mb区段(获得qKW4a的LOD为3.13，表型贡献率为9.49％，加性效应为1.74个籽粒，如图1B所示)，而且优异等位基因来源于SL41；基于玉米参考基因组序列信息(B73RefGen_v4)，在umc2038-umc1194发现了SL41和Y478间差异的10个SSR标记用于BC1群体的精细定位(图1C；表1)。标记开发使用SSRHunter software(motif长度≤4bp和重复数≥8bp)进行；将BC1群体的8种重组类型基因型自交(图1C)，自交果穗田间种植用于KW表型评价(图1C)。结果也将qKW4a定位于kw4–kw10区间(图1C；表1)。使用的引物序列见表2。
表1基于复合区间作图法的玉米百粒重QTL定位
Figure BDA0002905097490000031
表2 qKW4a精细定位与候选基因鉴定使用的引物
Figure BDA0002905097490000032
Figure BDA0002905097490000041
2、qKW4a的候选基因确认
qKW4a位于B73参考基因组chr4标记kw4–kw10间，物理距离为133kb，含有6个编码基因。对授粉后14天和28天的籽粒定位区段进行qRT-PCR(quantitative real-time PCR,qPCR)检测，结果显示：仅Zm00001d051976和Zm00001d051977在授粉后籽粒中表达。基于基因表达模式，仅Zm00001d051976在Ye478和SL41中差异表达，所以，将Zm00001d051976确认为qKW4a的候选基因(表3)；同时，也分析了Zm00001d051976在嵌套关联映射(Nestedassociation mapping,NAM)群体，以及在B73所有组织中的表达数据(http://www.qteller.com/)，表明Zm00001d051976属玉米籽粒充实期的功能基因。
表3 qRT-PCR在精细作图区间对百粒重进行鉴定
Figure BDA0002905097490000051
与相应同期同基因组的Ye478比较，经T测验，*p<0.05，**p<0.01；内参Actin(NM_001155179).n＝10；相对表达量计算采用ΔΔCt法。
利用表2中引物对Zm00001d051976和Zm00001d051977的基因组序列进行扩增，仅Zm00001d051976在Ye478和SL41间有序列差异(表2；图1D；图2)，而且在第1外显子和第2内含子区域SL41比Ye478有2个indel插入序列(图1D)，这可能是Ye478和SL41间KW差异的重要原因。
发明人还进行了2个关联群体Zm00001d051976区段的关联分析，结果显示位于Zm00001d051976区段具有4个KW显著关联的SNPs位点，而且具有较高表型贡献率(表4；图1E)，也进一步说明qKW4a候选基因确认的有效性。其中，候选基因区段的144个SNPs位点来源于489份自交系RNA转录组测序(覆盖全基因组总计556,809个SNPs)。489个玉米自交系KW表型评价来源3个环境(雅安；三亚；昆明；http://www.maizego.org/)。基于混合线性模型的最佳线性无偏预测(Best linear unbiased prediction,BLUP)进行3个环境的KW评价。使用TASSEL v4.2.1(MLM_Q+K)方法(Bradbury,P.J等，E.S.TASSEL:software forassociation mapping of complex traits in diverse samples.Bioinformatics,2007)进行BLUP和SNP间的关联分析。显著性阈值(p＝1/n，n＝144，-log10(p)＝2.16)(Fu,J.等,RNA sequencing reveals the complex regulatory network in the maizekernel.Nature Communications,2013)。通过线性回归估计PVE(Xu,R.Measuringexplained variation in linear mixed effects models.Statistics in Medicine,2003)；候选基因区段的41个SNPs位点来源于230份导入系(ILs；BC4F8；以Ye478为遗传背景(詹晶晶等，基于掖478导入系的玉米百粒重QTL鉴定.植物遗传资源学报，2015))，百麦克公司进行SLAF-GWAS测序(覆盖全基因组总计130,000个SNPs)。KW表型评价来源8个环境(保定和黑龙江，2013-2019)。进行8个环境的BLUP和SNP间关联分析，显著性阈值(p＝1/n，n＝45，-log10(p)＝1.65)。
表4 qKW4a候选基因的SNP与KW关联分析
Figure BDA0002905097490000052
Figure BDA0002905097490000061
3、qKW4a的候选基因功能分析
qKW4a候选基因为Zm00001d051976，编码Starch synthase V(SSV)，是首次基于图位克隆方法在玉米中确认的候选基因SSV，而且与玉米KW形成相关。淀粉分为直链淀粉和支链淀粉，是由α-1,4-葡聚糖和α-1,6-葡聚糖构成。淀粉合成酶(Starch synthase,SS)催化ADP-葡萄糖∷葡萄糖的葡萄糖基转移到现有葡聚糖链的非还原端(即：α-1,4-糖苷键)，从而使糖沉积在淀粉颗粒中。已报道植物的SS有5个亚家族，包括GBSS，SSI，SSII，SSIII和SSIV，而且每个亚家族的理化性质和结合底物，以及其功能均存在差异性。GBSS，SSI/SSII/SSIII和SSIV分别负责淀粉超长链合成，淀粉链延长和淀粉颗粒起始。SSV在拟南芥中被证明与淀粉合成相关，但候选基因SSV的功能在玉米中未见报道。本发明测定了双亲(SL41和Ye478)籽粒淀粉形成过程中的酶含量(表5)。因而qKW4a候选基因SSV属经图位克隆分离的新型与KW相关功能基因。
表5 SL41和Ye478的酶含量的生化检测
Figure BDA0002905097490000062
*Statistical significance at 0.05level；DAP，day after pollination，为授粉后的天数。
以上所述的实施例仅是对本发明的优选方式进行描述，并非对本发明的范围进行限定，在不脱离本发明设计精神的前提下，本领域普通技术人员对本发明的技术方案做出的各种变形和改进，均应落入本发明权利要求书确定的保护范围内。
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<110> 河北农业大学
<120> 玉米百粒重主效QTL位点qKW4a及其候选基因和应用
<160> 75
<170> SIPOSequenceListing 1.0
<210> 2
<211> 31293
<212> DNA
<213> 人工序列(Artificial Sequence)
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ttattgaaaa aaatcaggaa aaaagaagga tggtttcgca taccgctatt tgcagtgatg 60
gatcttggaa ggaagagcaa agaatgatat cagagcctgc atatattaaa tgtgccagag 120
cttcatcata tgcctcaaca aatgctattt tgtctccctg tttgagtaaa ccgaacaaat 180
gataaagaaa tgaaattgca ttattcatgt gacttgccat gattgtagaa agcctaatca 240
agttaaacac ggaagggtga tacctttatc tctcccttta atgcttgcaa aattgagttc 300
acaacagacc ctaacgtttc catgatgatg acctgtacat tataaactct tgcaactgtg 360
ctagcagaga aataaactgt ttaagtccca aggaagttat ctaacaaagt actctaccac 420
acctgagtat tcctctgcag agcaacatga actgcttctt taagactatg gacatctgaa 480
ttatcatcac agatacatcc tacctgacaa atggtgctta cctgatgtta cattcaatgt 540
agtaactaga taaacagaag tatgtatgca cagaacaaca gggtacatac tataatggat 600
gaatcaccat tgaaccgaag tctacgccta agtgcttgtc tgcaaatgga cttcccctcc 660
atatcattgg caaaatacct tcgaggaaga taaatgtatt tgaatggatc ccaaagttaa 720
ccatccaatc cataagatgc aaccaatatc ttctccctag aaaggattta atgcatctca 780
gaaaatctca gcaaatcaag tgccacttgc gacaagcgat ttcttagacc ctatgggaaa 840
gattatggaa agacaaacaa agaatgagct attttaccta tgtgcggcta atgttgattg 900
cagctcatgt cgtgaaccct gaataagagc atcttttgaa tgtgttgagg acatcaaaac 960
gactttattt gaataaacaa tgcctccctg catatacata agctaattga aagatatgtt 1020
gcacatacat aagttagtac aaatatatga tatactagct gagtgcccgt gcgttgcaac 1080
ggtaatatat aatatcagta tactacgata acttatatat aaaatgtatg ttatattatt 1140
atgagaaaat gtttcataat caatttgtga tcctagccat acataaattt tgttatttta 1200
atctagttgt ttcaccacta cattgcaacc atcagtatca tgcagacttc gatatatgct 1260
acgatttgta tggtctcatc attggagagc acgttccaca catgctggaa gaaatttcct 1320
cgtacatcgt tagtcatcag acacgcacca ccatacgctc ttgcttaaac aaaaaggtaa 1380
gtgtgtatgt ttgcgaagag aattaaaggc atgtcggcac aaaagctacc ctgacggtgg 1440
cgaggatgac gaactggtca ctgttgtcga tcctcctcta catcacctcc gacgctaaga 1500
tgacgccaca atcctcgata taatagtcgt cgaacgcgcg cgacatgacg agtaccgatg 1560
acccttggtt gggctaccaa acgaagtgca ccccggactc atcagcgact ccactacata 1620
catcatgttc gaggacactc acacaacgtc agcaacgttc atcgtcccag cgcacaagaa 1680
ttcatgtccg ttcagtagcg acttacgtgg taggttgggc tttaggtgga cgatgagcta 1740
gacgacgtga tggcgtcgtc gtccgatgcg gtgtccagaa caacccgaga gtcgtcaatg 1800
ttggcgacaa ccatgaggcc ccctgcttaa cgatggacag cgcggtgcag ctgctctgga 1860
ccgcgtccaa gtggtggctt cgcgggagct tgtcgtacac agcggcgcat gcgaccacgt 1920
aggactgctt ctagaggtcg aactgacagt cgccaagctt cttctcgtca tcgataaacg 1980
acgccaacgt gagtgtctcc tgctcatggt gtttgtttgg ggtttcaagt aagaaacatg 2040
gattcatgct tggcgttggt ttatgaaaat gactcagaag tctgatctat gaaaaaaata 2100
ttacgaagaa aaatgtcacg catgcaaaaa agggaattta agtataatac attattcaaa 2160
caaaagaaat tgcatgcaaa gctcttcttt aaataatatt acctccatcc aaaaatctaa 2220
ttcaagaatc tcggtgatac ttatctacta ctacgcattg tgcaagggta ccaagtgagc 2280
ttggtgagag atgtagtaga tatgttttct atcatagata tagacataaa cacaaatgtg 2340
gtgtagcggt agctactctt ggcctttgct tgagaggtca cggtttcaaa tccccttgag 2400
gacatgtgtt ttttttttaa ttttagctag acgtgggttg tacgggggaa tgagaatggg 2460
aatgagctct gcgggaggag ggaatgagaa agacagaata aagaatggga atgacaaaca 2520
tgagaatggg aacggtggca gaacgggaat ggcagaacag aaatggtggc agaacacggg 2580
aatggccgac tactcttaca gccttaatag tagtagagat aagttaatac aaagatatgt 2640
tcgatggata catcaaaaaa atttgaaaca attaacttcc atctggcatt tttaaacttg 2700
tattcgacaa aaatccaaac acgtttgaac aatatgtttc cagaaagaaa cagaattgcc 2760
ctcagtgcta aatgtttgtg ttggcagaga taacaacaat tcaggtacac gtctttaata 2820
tctaacttat attaaatcgc ataaaacctc cagttttaag atgatttacc ttaagaatat 2880
taacaagatg tgtcttgttt gtatcctgca gacgatcagg acgatgaagc ttacggggat 2940
caagcccaca catctccagt ttatttggct cctctaggca ctagaagcaa ttgcatgcaa 3000
cgtacaatat tttagagaaa aaaaacaaga aagcactaat agtgtagtag atacaggtac 3060
atataccgct gagccctaaa atggttttag atatatgcaa aatatattgc aatgattaga 3120
aacagtgacg ggtggaatga atcttaaaag tttgcatcct cttgaggtat attgacagta 3180
gttactgtgc atgtaacaac taacaactgg aaaaaaaaat catactgaat atttacttct 3240
gaattcaagt cttggcatgt cagtaatatc cgagtgttcc caagtccctg aatgtatcac 3300
aaaatttcaa gtcaaaactc atcaggactt caataacgaa atactaaagt acaaaagtaa 3360
gacctcaaaa ttgatttcag caaatacatt taaactcatc aggacttcaa taacgaaata 3420
ctaaagtaca aaagtaagac ctcaaaattg atttcagcaa atacatttgt aatgttgtat 3480
tattttatac aactgtaaac tcacaatttg gaatctaagt gagaacgcaa aagtcagggc 3540
catcctaaca cccaatttgc tttatcattt aggtcattaa tccagtctcc tgtgcacacc 3600
tctgaacatt acttttcaat aagtactata aaatgtaccg ttgaaagggg gggggtcact 3660
gggggcaagc ggaagtgccc aaacaaggtc aagttggcta agatagattc aagtttggtc 3720
ataatagact ggagaccttt tatctgtttt agataagggg actgatccct ctgtgaacat 3780
gcttaacaat ccagcataaa atttcagaac atgcacgaga aaaaacaaaa tatctaaaaa 3840
ataacagaaa aacagtaatc atgtctggaa tagatttaaa aaattctttt aaaaaatgta 3900
cgccaatcgc atcaggtgca gaatggtaaa atgcaagaca agtgccagat taaactccca 3960
ctgaaaatga aattaatcgt gtggcagttc atgtgggaaa aataccataa aatgtaatga 4020
atctaggaag tgatccttca atctaatata gcacttccac tcgtctgtcg ttagtgtgtg 4080
tgtctagtgg tctagttttg ctcttctgtg cctagtttta gtcgtctcgt cagtcgttca 4140
gcctcctact cttggttcgg acttcggagt gggtggtgct gtttaattgt aagaactcac 4200
ctttctacct taatacagtg ctacacatct ctcctgtgta ttcaagaaaa aaaatctaat 4260
atagaactta attttgtttg tattgaaaag aaaaacaagg tgcacatatt tcctcttttc 4320
cagaatgata tatttttcat tggtgatacg gacaaaatcc tgtcaaaagg tttatcatcc 4380
atttaacatg gagaataaag aacataggat tgcacaagac tgtttagcaa accattatca 4440
tgccattaaa atttaaaaac catagtatat ttgagaaagc aatgtgtttg agagagcata 4500
tcagtagtca taaatatata tacttaagta aaactgacct ggtgagcaaa tatttcccag 4560
aaaagcggtg ccactatagc agtttcccag ttatgtatgt gtataatgtc tggccgcttt 4620
ccagatttaa caatataatc cagggatgca cgggaaaaat aggaaaatct aaaaaagatg 4680
tagtaagaac aattaatctg tatgtctacg cttgcagcgc ttccaataaa aagctaatat 4740
aatgttctaa atgctaccat tattaacagt tactataaaa tatagtctgt tcaacatcag 4800
gttcttctgc atgttatatt aataaaagcc tgtcatgtca ctactcaaat ccaatactct 4860
tacataccgc tcaaagtcat cagggtagcc acgtaacata tcacgactaa aataggagag 4920
ctgagatggt tctattatta caagaccaac accacttgac gtactgcata aaagaagcaa 4980
aattaagagt gaaatgcctt tagaaataca caaatacaga agtgtcagct aaaataaaag 5040
aacgagataa taggcttgtt acatctcttt gcacactagg agtgtatctt tacccagtcc 5100
atattttatt tttatgccaa aggccaccga aataagattc atattctgcc tcagatctcc 5160
ttagaccatg aattccatcc aagtttatgc tggtgtacct attttgtgtc ataccaatgc 5220
gatgagcatg atagctttgc atgcaataag caagatgtaa atgtggcaca agccaaaaca 5280
atccatatta ctaaggataa caacgaattt caaatgtaac acatgtgatc aagaacagga 5340
agagaaaaaa attgaggagt acataccaaa acaatactgc aaaacataac ttgtttcaaa 5400
acagtttggt aattaaatca aatcatttgg gccgtgctgt cttgatggaa taaacaacac 5460
aattaagact agattgccat caaaactaaa agttgacaat caaattcaaa ccttcaggaa 5520
aatgcccagt ttagagcatc cgtctagaat gaacaaaaaa ttgtctcgtc tgcacattga 5580
tatttaaatt tcaggattct actcttacat agtaatagct aaaaggaagc ttagaagcgt 5640
gttacatact tgtattcttg aacatttttt acattaaatc aacccaaaag aaacttcccc 5700
attcatatga ctaattggac cactacaaca taaaagctca aggtgtccag acagtctatc 5760
aagcattatg taccaaaggt tcggttagtg aagagaggtt gcatacttgg gtagtatcac 5820
ctccaccaag ttaccctttt tttgaatctc acaagataaa ctggcaacat atgctgataa 5880
tggtccacat gatgctactg ggtacatttc actacagatg tgcacaatat gaagaggttt 5940
cctgcattga acataggctc aaatatagca tcaacaaata tgataaaaca agacaaatta 6000
aatatgtcgc atgattatta taaagtaagt gcagatacac taggtgaaat atcagcggga 6060
agcagtggcg gggccaaaaa ttatacatag ggaggcagcc aaaagattca aaacataggc 6120
aagaatagtg ttaccaggac gcaggtgtgg gaaccagaat ccgatacaga tgcaggtgtt 6180
tagctaggca ggctcagaaa aataggattc aggaattctt ctaatacatt ttaaaaaata 6240
gaacagcatt ttagcccttc aaatattaag tacaatatta atccctcttc ttagtaaaat 6300
gacatgaaac tctcctatgt cgttaaaaaa aatctaagat taatcctatt tactataaaa 6360
caaactaaaa ggacactaaa gattccattg tgtgctttac caaaggaaaa gacctgggat 6420
tacagcaaaa gctccagatt agctacagtt tgtgcataga acaacagtca aaactagcat 6480
ttaaagtgat tccctctcta tggctacaca cctatcattg tcttaatata accagcattt 6540
aaagtgattc cctctctacg gctacacaca ttttcattgt cttttgccat ttttccctgt 6600
catggataac acacatgagc ttcctcttgc cctctgcagc aagattcttt ctcaatgtat 6660
cagatatgag ccaggccatg tcaggtgtca atgggaaatt cgttgtgtca agttgtcatg 6720
gcagcaataa aaaatggaca taggtatccc gcaacactgc agaaaatttg agtaaaggcg 6780
gtaaaactgg aaaaaaaagt aaggtaagtg gtagcatact ctattggttt acgcaaaaag 6840
agttgcagtt cagacaacaa ttcaatattt ggcttatgat gaatatcaga aaaggtgttt 6900
tgtattaggc ttctattagt gacaagtttc tctcttaaat cagatgcttc cgtcatgctg 6960
atcattccag aaatgaccat tgtatctatc cgcaggagta actcactgaa acttgacgac 7020
tgtgctgctt ctaggggaat tcttgaaagg agtcccatga gaaaatggaa tatgtagaag 7080
tatagcgaaa ggtatatgaa aaaatatttg atgcatgtag tttacctcta tagttaagaa 7140
tgagaaatgg aaagtaaaac aatcatatta acaatcagca taggattgct gaagattgag 7200
aaatttcaca ttaccatcca tatagttcct aagcctgaaa atgttggcaa ccgagaccgc 7260
accttgcatt tctgtttcta gtccttgtat ttcctcgaga agcaaattat tttcatcctg 7320
cagtttcttc atctcgtcca tggcaataag ccgctgcttg ttaatgtaca ggatatctgt 7380
aacagacaaa tgttaagacc tgtttatcag caaggaagga agcgatatga aagacaggtt 7440
gaaatggaaa aggtgctaca ttatgatgaa aaataatagt tgtggtacta agtgttgtcc 7500
attttacgaa ttttcacttt attaaaaaaa aacaaagggg ccgactgggt gggccgctac 7560
taggctggag gccgagcagc ccaccggcaa gcgctggtaa ataaccagct agggcacctc 7620
ctgcctgtgc gtcaaaaagg gacagagagc ggcggcgaag gcacaacggc ttggccggcc 7680
tgcacggccg cctcttgtgt ttctctctgt tcaacggcct ccgaatttgg tgcccggtct 7740
gttccataag tatgggtgtg tttcccctct ctctggcgtt ggctgagcag tcggatgttc 7800
ttcttcctcc ctctgtgtgt gctgctccgg taagaactca aactaccagt tcgtttcctg 7860
gtattgtggt ggcgattctt gctgggagtg gttcaaagtg ggaggaagtg gtgctgtgct 7920
tgcgcagtgt gagttacctg tgctttgttg ccggaatcgt gcacttctcc ggtgctctga 7980
ttcttgtctg atctctgtcc accgtggcct ggtgcttggg ctccctctgc gtggtgttcc 8040
tcttctcggg ctggtgccgt ggacgtctcc ctcggttccg gcagcggctc tagtatctcc 8100
gtcaacctcc cgccgtgcag gcccggacgt gggcggcgga attggttttc tgggacagaa 8160
cttcgtgttc gctgagctga tcttcccacg tagacaatct acgtgctgtg agttattcgt 8220
tgcctcactt tatttactgt ttaattttgt gcaatctgct agtattaaat acttgggtgt 8280
gattgcactg ttgtggacgg atttgtgtgc tcgatgatga tagcggtgta gtgagacaac 8340
gtgacagtct gggttttagt gtattatcat tttcagcatt gatattattt gtgcctaatt 8400
aaatctctct ttgttaaggc tgtttacctg tgttaaattt tgttaattta ttaagtacac 8460
agctcctttc tatcagtatt tatatttgat tattaggctt attacatctg atctaaaacc 8520
cttatttggt aagataggac tgtcacgtta gtcgaggatc tctgatcacc ttttagcgta 8580
ggagggctgt ttattacagc aacccgctat tttgagagca atattttaag ataaacaatt 8640
atgtttattt gtttaatctg ccttcttcat ctgcaagcca tgtttaatta tgaaatggac 8700
ctgagtagct attttacatg tttacatgtt ttagaattgc tactgttcat tgctgcaata 8760
attacatatg gcatttatct gagttatatg cttgttttat tcggaattag aatatttaat 8820
ttatttaaat atgaaggaag catgtcactt atttctctaa ttttacaaca atccagaaaa 8880
ccaattcgta taatggcctc cagttctgat gtcattaagc ctgaagcgtt cgacggcgca 8940
agctttaagc gctggcagat taagactcgc atgtggctca ctgacctaaa attattttgg 9000
gtagtgacgt cggctgttcc ccaggctgca tcagatgatt ctgatgatgc agcgaaggcc 9060
gccgcactag cggagaaggc caagtgggac gaagccaatg aggcatgctt gtctcgtctg 9120
ctgaacgtct tgtcgaaccg cttatttgac gtgtactctg gttttacctc cgctaagggt 9180
ctatggactg aacttgagaa cgagttctct gaagttgaca atggcaatga gtcattcaca 9240
acggaaaact accttaacta taaaatggtt gagggaaggt ctgttatgga gcagctacag 9300
gagattcaac tccttgttag ggacttggtc caatatggct gcgtcctacc agacagtttt 9360
caggtgaatg caatactggc aaagctcccg ccttcttggc gagactttgt gacttcacgc 9420
cgtcacatga aaaagcaaat gactctcact gagctgtcag ctgcgataaa tgtggaagag 9480
cgtgcaaggt ccagtaacaa gccatcccag caactccagg ctcacgttgt tgagaagggt 9540
ggagatagaa aattccagaa gaagaagaag aactctccac agaagaatct gaaccaacct 9600
aaatccaaaa agatgaagaa gaaaaaggag gactttatct gctacgtttg tggtgtctcc 9660
gggcatacag ctcggaggtg caaacttagg aagggaaaag gtcctccacc tcagcgcaaa 9720
gaagggaacg tggtggttaa ctccacccca gggtatgcac ttcaggcctt tatggcaagt 9780
ccatcagatg actggtggat ggattccggt gctactgttc atatttgtgc tgatcgatcc 9840
atgttctctt cattccaggg atacaacagc gcaccagtct tgatgggaaa tggtgttccg 9900
gcagcagttc gaggtactgg acaggtctac ttgaagttaa cttcaggaaa gacactcgtg 9960
ctgaaggacg tactctatgt gccctcgatg agccggaacc tcatttcagt gtcgctgcta 10020
tgtcgacagg gtctcaaatt agtgtttgag tctaataaag tggtcctgtc taagtttggt 10080
acttttgttg gaaagtcata tgagtcaggc ggtttgttcc gtctttctgt tttgaacaat 10140
cattcttctt accatgttaa tgtggtctgt aataacgatt ccattaataa tatatggcat 10200
tcccgtttgt gtcatgtgaa ttttgaggcc attaagagat taagtgacat gtctttaatt 10260
cctgaatata aacatgttaa aggtgtaaaa tgtggtattt gtgtgcaagc taagcagcct 10320
agaaaaccgt ttcatacggt tgaaggcaga agtaccactc ctttagaact aattcactca 10380
gatatctgtg agatgaatgg tataatcaca aaaggcggta aaagatactt tcttaccttg 10440
atagatgatg ctaccaggtt ctgttatatt tacttactca gaactaaaga tgaggcacta 10500
gaacacttta agatctataa gactgaagtt gaaaaccagt tagacaagaa aattaaaagg 10560
ttacggtctg atagaggagg tgaatacctt tccaaccttt ttgatgagta ctgcaaagag 10620
tgtggaatca ttcatgaaac caccgctcca tattcacctc agtcaaatgg ggtagctgaa 10680
aggaagaacc gaacggtgtg tgacttagct aatgctttgt tgcaaagttc ggggatgcct 10740
gacatctggt ggggcgaggc agtactcaca gtgtgctatg tcctgaacag ggtgccgcct 10800
cgcaaccgtg aggccacgcc ctatgaagga tttaaaggaa ggaagccaga tctttcgcat 10860
cttcggactt ggggctgcct tgcaaaggtg aatgtcccgt tgccgaagaa gagaaagcta 10920
ggccctaaga ccgtagactg tgtcttcctg ggttatgcac ataacagtgc agcttataga 10980
tttctagtgg tccattccga gacaagcgaa atcgctataa atgtaataat ggagtctcgg 11040
gatgtgacat tctttgagag catcttccct atgcgggata aggaagtagt agccccagat 11100
ggtccatctc ggacatattc tctgccctcg agtgtcaatg accagactcc agatttggag 11160
ttaaggagga gtaagagaca aaggactgaa aagtctctgg gtgatgatta tattatttat 11220
ctagtggatg aagaaccacg ctccctcaca gaggcatata catctctgga tgcagagtac 11280
tggagggagg ctgtccgtag cgagatggat tcaatcatct cgaacggaac ttgggagata 11340
actgatctcc cagctggttg caagcccgtg ggctgtaaat ggatctttag gaggaagagg 11400
agacccgatg gtactattga aaagtacaag gcccgtcttg tggctaaggg ttttactcaa 11460
aagaaagagg aggattattt tgatacttat tctccggtgg ctcgattgcc cacaatccgt 11520
gtgcttttag cactggcagc tgcttataaa cttcttgtcc atcaaatgga cgtaaagaca 11580
gcattcctaa atggagagct ggaagaggaa atttatatgc agcaaccaga aggatttgtg 11640
gttaaaggtc aagagagcaa agtgtgtcgc ttgattaaat ccttatacgg tcttaagcaa 11700
gctcccagac aatggcatga gaagtttaat aatactctga ccactgccgg gttttgtgta 11760
aacgaagccg acaagtgtgt gtattatcgc ttttctgggg gcaaaggtgt catcatgtgt 11820
ttatacgttg atgacatatt gatatttggg accgatttgg aggctatcat ggagacaaaa 11880
atattcctct ccaagaactt tgatatgaag gacttgggtg aagctgatgt aatattgaac 11940
atcaagctaa taaagggtga ggataggatt actttgtcac aatcccatta tgtggaaaag 12000
gtcctgactc gctttggaca tatggactgt aaaccagtca ccacgcccta tgatccatcc 12060
tacacgctca gtaaatatga aggcgagcct gtgaaccagc tactatattc gcagatcatc 12120
ggatctctca tgtacctgag cagtgcaact agaccagata tctcatatgc agtatgcaga 12180
ctggctcgtt attcggccag tccaggagat agacactggg tagccttgta cagagtgctt 12240
cggtatttga agggagcgat gaatcttggt attaaatata ccggatttcc gtcagtactt 12300
gaaggattca gtgatgcaaa ctggatctct gactcggatc aaatgaagtc tacaagtggc 12360
tatgtgttca ctttagctgg tggggccgtg tcatggagat cgtctaaaca atcagtgtcg 12420
acacgttcca ccaaggaggc tgaattagtt gcacttgatt cagcagcatt ggaggctgaa 12480
tggttgagag acctgttgtc agaccttcca atgctagcaa agccgatacc ggctgtccta 12540
gtatactgtg acaacacttc tgtgttgctg aaagtgaaca gtagaaagga caaccaaaaa 12600
tcctcgaggc atatcagaag acgacttgat tcatgtcggc atgctagaga gacgggtgta 12660
ataaccgtag attacatcaa gtctgaaagg aaccttgctg atcctttcac caaaggactg 12720
gctcaaaagc caatccaagc agcgtgcatg ggaatgggac tcgtaccctg ttagcggttt 12780
caactgcgga taactgtcct gtgtgaccgg agatcccgag atccaggctc caggaaacga 12840
agcactgtga aaccaagagc acaaaagata aagagtctca tgagctgccg tgctctgtct 12900
gtatggcagg ttgagcgata tgctcttaat gaagtcaatg ctataagcag ggaaattgtc 12960
ctacagaatt tccttaacga tttcacctat atgagctgac tgtggggtcg cagtccagga 13020
gagaatgggg ttttctctct agtgagctca tgaatagata ttaaagggca tgaccgtaat 13080
gccctgcccc gtaagagaag cagataatct gcctagtact atgtgccttt tgtgcgaaga 13140
ttctcacact agggtttgtg gatcaaggcg tagtcctccg cttactcctg caggggtgga 13200
gtacactggg ataggctttg gttcaaacct aacaagtacc tctgccgaaa agtagtatat 13260
aaacagtacg ggagctcaat gacattgatc agaaataaga gtgaaaatgg tggggattgt 13320
tgtccatttt acgaattttc actttattaa aaaaaacaaa ggggccgact gggtgggccg 13380
ctactaggct ggaggccgag cagcccaccg gcaagcgctg gtaaataacc agctagggca 13440
cctcctgcct gtgcgtcaaa aagggacaga gagcggcggc gaaggcacaa cggcttggcc 13500
ggcctgcacg gccgcctctt gcgtttctct ctgttcaacg gcctccgaat ttggtgaccg 13560
gtttgttcca taagtatggg tgtgtttccc ctctctctgg cgttggctga gcagtcggat 13620
gttcttcttc ctccctctgt gtgtgttgct ccggtaagaa ctcaaactac cagttcgttt 13680
cctggtattg tggtggcgat tcttgctggg agtggttcaa agtgggagga agtggtgctg 13740
tgcttgcgca gtgtgagtta cctgtgctct gttgccggaa tcgtgcactt ctccggtgct 13800
ctgattcttg tctgatctct gtccaccgtg gcctggtgct tgagctccct gctgcgcggt 13860
gttcatcttc tcgggctggt gccgtggacg tctccctcgg ttccggcagc ggctctagta 13920
tctccgtcaa cctcccgccg tgcaggcccg gacgtgggcg gcggaattgg ttttctggga 13980
cagaacttcg tgttcgctga gctgatcttc ccacgtagac aatctacgtg ttgtgagtta 14040
ttcgttgcct cactttattt actgtttaat tttgtgcaat ctgctagtat taaatacttg 14100
ggtgtgattg cactgttgtg gacggatttg tgtgctcgat gatgatagcg gtgtagtgag 14160
acaacgtgac agtctgggtt ttagtgtatt atcattttca gcattgatat tatttgtgcc 14220
taattaaatc tctctttgtt aaggttgttt acctgtgtta aattttgtta atttattaag 14280
tacacagctc ctttctatca gtatttatat ttgattatta ggcttattac atctgatcta 14340
aaacccttat ttggtaagat aggactgtca cgttagtcga ggatctctga tcacctatta 14400
gcgttggagg gctgtttatt acagcaacct gctattttga gagcaatatt ttaagataaa 14460
caattatgtt tatttgttta atctgccttc ttcatctgca agccatgttt aatttccaaa 14520
tggacctgtg tagctatttt acatgtttac atgtcttaga attgctactg ttcattgctg 14580
ttataattac atatggcatt tgtctgagtt atatgcttgt tttattcgga attagaatat 14640
ttaatttatt taaatatgaa ggaaacatgt cacttatttc tctatattta caacactaag 14700
ttgtggtctg atacaaatac agaacattaa cattatgtga agcattaatg tgaagcaagg 14760
ggagagtaat tactccgttg agccaggttg aagagctctg taatatcacc ctcacgttgc 14820
tcgtcagcca aacttggtac ctggaaacag caattaccct cataagcctc ttaaacgtaa 14880
accaggaaag caaataaaaa cagaaaataa gtggaattgt ctgtggtgat gttatccatg 14940
cctgatcgct atctgcttcg acgtccaacc tacaaaattc ccagcacccc gacatatata 15000
aacatcatcc gctggtttaa tccaattcga gcgctaataa aatcgagatg tagagggaac 15060
ccaccggacg gaaccggagc tctcgcggcg ggccttacca tgagaatcgc cgttcccctc 15120
cctgagacac gaattagagg accgaaatca gctgaggaac aggcacgcat acagagtaca 15180
gtactacagt tacacacccg cgcgcagcaa ttatgggagc acgatcgcac ggagtggtgc 15240
tgggtgtggg atttgcacgc acctggaaga ggagtaggag gagcaccggg ttagaagcac 15300
cggtggagca gcgcgggcgg agggggagca atgtgccccg ccgcacggac gcaggatacg 15360
ccgtcgctgt gatcgaggag ctggtggcgg ggcccgcgac gccgcaccga ccacgaggcg 15420
cggcgtctcc atggttgcct gctacgcacg ctgctcgtac gcctccttgg ttagctatgc 15480
gcgccgtacc tctccgcgga ttgcccaggt gaaagagaca attccgtgtg cttcaatggc 15540
cagaggcgag gctgctgcct gatggcggct ggcgggcgct gagccgctgc gctgatcggt 15600
gcgaggaggc gccgttcggc gtgcgtgctc gcggaggtgg gacgtggcac gtgggcttat 15660
tgaaaaaaat caggaaaaaa gaaggatggt ttcgcatacc gctatttgca gtgatggatc 15720
ttggaaggaa gagcaaagaa tgatatcaga gcctgcatat attaaatgtg ccagagcttc 15780
atcatatgcc tcaacaaatg ctattttgtc tccctgtttg agtaaaccga acaaatgata 15840
aagaaatgaa attgcattat tcatgtgact tgccatgatt gtagaaagcc taaacaagtt 15900
aaacacggaa gggtgatacc tttatctctc cctttaatgc ttgcaaaatt gagttcacaa 15960
cagaccctaa cgtttccatg atgatgacct gtacattata aactcttgca actgtgctag 16020
cagagaaata aactgtttaa gtcccaagga agttatctaa caaagtactc taccacacct 16080
gagtattcct ctgcagagca acatgaactg cttctttaag actatggaca tctgaattat 16140
catcacagat acatcctacc tgacaaatgg tgcttacctg atgttacatt caatgtagta 16200
actagataaa cagaagtatg tatgcacaga acaacagggt acatactata atggatgaat 16260
caccattgaa ccgaagtcta cgcctaagtg cttgtctgca aatggacttc ccctcaatat 16320
cattggcaga ataccttcga ggaagataaa tgtctttgaa tggatcccaa agttcaccat 16380
ccaatccata agatgcaacc aatatcttct ccctagaaag gatttaatgc atctcagaaa 16440
atctcagcaa atcaagtgcc acttgcgaca agcgatttct tagaccctat gggaaagatt 16500
atggaaagac aaacaaagaa tgagctattt tacctatgtg cggctaatgt tgattgcagc 16560
tcatgtcgtg aaccctgaat aagagcatct tttgaatgtg ttgaggacat caaaacgact 16620
ttatttgaat aaacaatgcc tccctgcata tacataagct aattgaaaga tatgttgcac 16680
atacataagt tagtacaaat atatgatata ctagctgagt gcccgtgcgt tgcaacggta 16740
atatataata tcagtatact acgataactt atatataaaa tgtatgttat attattatga 16800
gaaaatgttt cataatcaat ttgtgatcct agccatacat aaattttgtt attttaatct 16860
agttgtttca ccactacatt gcaaccatca gtatcatgca gacttcgata tatgctacga 16920
tttgtatggt ctcatcattg gagagcacgt tccacacatg ctggaagaaa tttcctcgta 16980
catcgttagt catcagacac gcaccaccat acgctcttgc ttaaacaaaa aggtaagtgt 17040
gtatgtttgc gaagagaatt aaaggcatgt cggcacaaaa gctaccctga cggtggcgag 17100
gatgacgaac tggtcactgt tgtcgatcct cctctacatc acctccgacg ctaagatgac 17160
gccacaatcc tcgatataat agtcgtcgaa cgcgcgcgac atgacgagta ccgatgaccc 17220
ttggttgggc taccaaacga agtgcacccc ggactcatca gcgactccac tacatacatc 17280
atgttcgagg acactcacac aacgtcagca acgttcatcg tcccagcgca caagaattca 17340
tgtccgttca gtagcgactt acgtggtagg ttgggcttta ggtggacgat gagctagacg 17400
acgtgatggc gtcgtcgtcc gatgcggtgt ccagaacaac ccgagagtcg tcaatgttgg 17460
cgacaaccat gaggccccct gcttaacgat ggacagcgcg gtgcagctgc tctggaccgc 17520
gtccaagtgg tggcttcgcg ggagcttgtc gtacacagcg gcgcatgcga ccacgtagga 17580
ctgcttctag aggtcgaact gacagtcgcc aagcttcttc tcgtcatcga taaacgacgc 17640
caacgtgagt gtctcctgct catggtgttt gtttggggtt tcaagtaaga aacatggatt 17700
catgcttggc gttggtttat gaaaatgact cagaagtctg atctatgaaa aaaatattac 17760
gaagaaaaat gtcacgcatg caaaaaaggg aatttaagta taatacatta ttcaaacaaa 17820
agaaattgca tgcaaagctc ttctttaaat aatattacct ccatccaaaa atctaattca 17880
agaatctcgg tgatacttat ctactactac gcattgtgca agggtaccaa gtgagcttgg 17940
tgagagatgt agtagatatg ttttctatca tagatataga cataaacaca aatgtggtgt 18000
agcggtagct actcttggcc tttgcttgag aggtcacggt ttcaaatccc cttgaggaca 18060
tgtgtttttt ttttaatttt agctagacgt gggttgtacg ggggaatgag aatgggaatg 18120
agctctgcgg gaggagggaa tgagaaagac agaataaaga atgggaatga caaacatgag 18180
aatgggaacg gtggcagaac gggaatggca gaacagaaat ggtggcagaa cacgggaatg 18240
gccgactact cttacagcct taatagtagt agagataagt taatacaaag atatgttcga 18300
tggatacatc aaaaaaattt gaaacaatta acttccatct ggcattttta aacttgtatt 18360
cgacaaaaat ccaaacacgt ttgaacaata tgtttccaga aagaaacaga attgccctca 18420
gtgctaaatg tttgtgttgg cagagataac aacaattcag gtacacgtct ttaatatcta 18480
acttatatta aatcgcataa aacctccagt tttaagatga tttaccttaa gaatattaac 18540
aagatgtgtc ttgtttgtat cctgcagacg atcaggacga tgaagcttac ggggatcaag 18600
cccacacatc tccagtttat ttggctcctc taggcactag aagcaattgc atgcaacgta 18660
caatatttta gagaaaaaaa acaagaaagc actaatagtg tagtagatac aggtacatat 18720
accgctgagc cctaaaatgg ttttagatat atgcaaaata tattgcaatg attagaaaca 18780
gtgacgggtg gaatgaatct taaaagtttg catcctcttg aggtatattg acagtagtta 18840
ctgtgcatgt aacaactaac aactggaaaa aaaaatcata ctgaatattt acttctgaat 18900
tcaagtcttg gcatgtcagt aatatccgag tgttcccaag tccctgaatg tatcacaaaa 18960
tttcaagtca aaactcatca ggacttcaat aacgaaatac taaagtacaa aagtaagacc 19020
tcaaaattga tttcagcaaa tacatttaaa ctcatcagga cttcaataac gaaatactaa 19080
agtacaaaag taagacctca aaattgattt cagcaaatac atttgtaatg ttgtattatt 19140
ttatacaact gtaaactcac aatttggaat ctaagtgaga acgcaaaagt cagggccatc 19200
ctaacaccca atttgcttta tcatttaggt cattaatcca gtctcctgtg cacacctctg 19260
aacattactt ttcaataagt actataaaat gtaccgttga aagggggggg gtcactgggg 19320
gcaagcggaa gtgcccaaac aaggtcaagt tggctaagat agattcaagt ttggtcataa 19380
tagactggag accttttatc tgttttagat aaggggactg atccctctgt gaacatgctt 19440
aacaatccag cataaaattt cagaacatgc acgagaaaaa acaaaatatc taaaaaataa 19500
cagaaaaaca gtaatcatgt ctggaataga tttaaaaaat tcttttaaaa aatgtacgcc 19560
aatcgcatca ggtgcagaat ggtaaaatgc aagacaagtg ccagattaaa ctcccactga 19620
aaatgaaatt aatcgtgtgg cagttcatgt gggaaaaata ccataaaatg taatgaatct 19680
aggaagtgat ccttcaatct aatatagcac ttccactcgt ctgtcgttag tgtgtgtgtc 19740
tagtggtcta gttttgctct tctgtgtcta gttttagtca tctcgtcagt cgttcagcct 19800
cctactcttg gttcggactt cggagtgggt ggtgctgttt aattgtaaga actcaccttt 19860
ctaccttaat acagtgctac acatctctcc tgtgtattca agaaaaaaaa tctaatatag 19920
aacttaattt tgtttgtatt gaaaagaaaa acaaggtgca catatttcct cttttccaga 19980
atgatatatt tttcattggt gatactagga caaaatcctg tcaaaagctt tatcatccat 20040
ttaacatgga gaataaagaa cataggattg cacaagactg tttagcaaac cattatcatg 20100
ccattaaaat ttaaaaacca tagtatattt gagaaagcac tgtgtttgag agagcatatc 20160
agtagtcata aataaatata cttaagtaaa actgacctgg tgagcaaata tttcccagaa 20220
aagcggtgcc actatagcag tttcccagtt atgtatgtgt ataatgtctg gccgctttcc 20280
agatttaaca atataatcca gggatgcacg ggaaaaatag gaaaatctaa aaaagatgta 20340
gtaagaacaa ttaatctgta tgtctacgct tgcagcgctt ccaataaaaa gctaatataa 20400
tgttctaaaa tgctaccatt attaacagtt actataaaat atagtcagtt caacatcagg 20460
ttcttctgca tgttatatta ataaaagcct gtcatgtcac tactcaaatc caatactctt 20520
acataccgct caaagtcatc agggtagcca cgtaacatat cacgactaaa ataggagagc 20580
tgagatggtt ctattattac aagaccaaca ccacttgacg tactgcataa aagaagcaaa 20640
attaagagtg aaatgccttt agaaatacac aaatacagaa gtgtcagcta aaataaaaga 20700
acgagataat aggcttgtta catctctttg cacactagga gtgtatcttt acccagtcca 20760
tattttattt ttatgccaaa ggccaccgaa ataagattca tattctgcct cagatctcct 20820
tagaccatga attccatcca agtttatgct ggtgtaccta ttttgtgtca taccaatgcg 20880
atgagcatga tagctttgca tgcaataagc aagatgtaaa tgtggcacaa gccaaaacaa 20940
tccatattac taaggataac aacgaatttc aaatgtaaca catgtgatca agaacaggaa 21000
gagaaaaaaa ttgaggagta cataccaaaa caatactgca aaacataact tgtttcaaaa 21060
cagtttggta attaaatcaa atcatttggg ccgtgctgtc ttgatggaat aaacaacaca 21120
attaagacta gattgccatc aaaactaaaa gttgacaatc aaattcaaac cttcaggaaa 21180
atgcccagtt tagagcatcc gtctagaatg aacaaaaaat tgtctcgtct gcacattgat 21240
atttaaattt caggattcta ctcttacata gtaatagcta aaaggaagct tagaagcgtg 21300
ttacatactt gtattcttga acatttttta cattaaatca acccaaaaga aacttcccca 21360
ttcatatgac taattggacc actacaacat aaaagctcaa ggtgtccaga cagtctatca 21420
agcattatgt accaaaggtt cggttagtga agagaggttg catacttggg tagtatcacc 21480
tccaccaagt tacccttttt ttgaatctca caagataaac tggcaacata tgctgataat 21540
ggtccacatg atgctactgg gtacatttca ctacagatgt gcacaatatg aagaggtttc 21600
ctgcattgaa cataggctca aatatagcat caacaaatat gataaaacaa gacaaattaa 21660
atatgtcgca tgattattat aaagtaagtg cagatacact aggtgaaata tcagcgggaa 21720
gcagtggcgg ggccaaaaat tatacatagg gaggcagcca aaagattcaa aacataggca 21780
agaatagtgt taccaggacg caggtgtggg aaccagaatc cgatacagat gcaggtgttt 21840
agctaggcag gctcagaaaa ataggattca ggaattcttc taatacattt taaaaaatag 21900
aacagcattt tagcccttca aatattaagt acaatattaa tccctcttct tagtaaaatg 21960
acatgaaact ctcctatgtc gttaaaaaaa atctaagatt aatcctattt actataaaac 22020
aaactaaaag gacactaaag attccattgt gtgctttacc aaaggaaaag acctgggatt 22080
acagcaaaag ctccagatta gctacagttt gtgcatagaa caacagtcaa aactagcatt 22140
taaagtgatt ccctctctat ggctacacac ctatcattgt cttaactagc atttaaagtg 22200
attccctctc tacggctaca cacctatcat tgtcttctgc catttttccc tctcatggat 22260
aacacacatg agcttcctct tgccctctgc agcaagattc tttctcaatg tatcagatat 22320
gagccaggcc atgtcaggtg tcaatgggca attcgctgtg tcaagttgtc atggcagcaa 22380
taaaaaatgg acataggtat cccgcaacac tgcagaaaat ttgagtaaaa ggcggtaaaa 22440
ctgggaaaaa aaagtaaggt aagtggtagc atactctatt ggtttacgca aaaagagttg 22500
cagttcagac agcaattcag tatttggctt atgatgaata tcacaaaagg tgctttgtat 22560
tatgcttcta ttagtgacaa gtttctctct taaatcagat gcttccgtca tgctgatcat 22620
tccagaaatg accattgtat ctatccgcag gagtaactca ctgaaacttg acgactgtgc 22680
tgcttctagg ggaatacctg aaaggagtcc catgagaaaa tggaatatgt agaagtatag 22740
cgaaaggtat atgaaaaaat atttgatgca tgtagtttac ctctatagtt aagaatgaga 22800
aatggaaagt aaaacaatca tatcaacaat cagcatagga ttgctgaaga ttgagaaatt 22860
tcacattatc atccatatag ttcctaagcc tgaaaatgtt ggcaaccgag accgcacctt 22920
gcatttctgt ttctagtcct tgtatttcct cgagaagcaa attattttca tcatgcagtt 22980
tcttcatctc ctccatggca ataagccgct gcttgttaat gtacaggata tctgtaacag 23040
acaaatgtta agacctgttt atcagcaagg aaggaagcga tatgaaagac aggttgaaat 23100
ggaaaaggtg ctacattatg atgaaaaata atagttgtgg tactaagtgt tgtccatttt 23160
acgaattttc actttattaa aaaaaaacaa aggggccgac tgggtgggcc gctactaggc 23220
tggaggccga gcagcccacc ggcaagcgct ggtaaataac cagctagggc acctcctgcc 23280
tgtgcgtcaa aaagggacag agagcggcgg cgaaggcaca acggcttggc cggcctgcac 23340
ggccgcctct tgtgtttctc tctgttcaac ggcctccgaa tttggtgccc ggtctgttcc 23400
ataagtatgg gtgtgtttcc cctctctctg gcgttggctg agcagtcgga tgttcttctt 23460
cctccctctg tgtgtgctgc tccggtaaga actcaaacta ccagttcgtt tcctggtatt 23520
gtggtggcga ttcttgctgg gagtggttca aagtgggagg aagtggtgct gtgcttgcgc 23580
agtgtgagtt acctgtgctt tgttgccgga atcgtgcact tctccggtgc tctgattctt 23640
gtctgatctc tgtccaccgt ggcctggtgc ttgggctccc tctgcgtggt gttcctcttc 23700
tcgggctggt gccgtggacg tctccctcgg ctctggcagc gactctagta tctccgtcaa 23760
cctcccgccg tgtaggcccg gacgtgggcg gcggaattgg ttttctggga cagaaccttg 23820
tgtttgctga gctggtcttc ccacgtagac aatctacgtg atgtgagtta ttcgttgtct 23880
cactttattt actgtttaat tttgtgcaat ctgctagtat taaatacttg ggtgtgattg 23940
cactgttgtg ggcggatttg tgtgctcgat gatgatagcg gtgtagtgag acaacgtgac 24000
agtctgggtt ttagtgtatt atcattttca acattaatat tatttgtgcc taattaaatt 24060
tctctttgtt gaggctgttt acctgtgtta aattctgtta ttttatttag tacacagctc 24120
cttagtatta gtatttagat ttgattatta ggcttattac atctgatcta aaaccctgat 24180
ttggtaagat agggctgtca cgttagtcca cctattagcg taggagggct atttattaca 24240
gcaacccgct tttttgagag caatatttta agataaacaa ttatgtttat ttgtttaatc 24300
tgccttcttc atctgcaagc catgtttaat tatgaaatgg acctgagtag ctattttaca 24360
tgtttacatg ttttagaatt gctactgttc attgctgcaa taattacata tggcatttat 24420
ctgagttata tgcttgtttt attcggaatt agaatattta atttatttaa atatgaagga 24480
agcatgtcac ttatttctct aattttacaa caatccagaa aaccaattcg tattatggcc 24540
tccagttctg atgtcattaa gcctgaagcg ttcgacggcg caagctttaa gcgctggcag 24600
attaagactc gcatgtggct cactgaccta aaattatttt gggtagtgac gtcggctgtt 24660
ccccaggctg catcagatga ttctgatgat gcagcgaagg ccgccgcact agcggagaag 24720
gccaagtggg acgaagccaa tgaggcatgc ttgtctcgtc tgctgaacgt cttgtcgaac 24780
cgcttatttg acgtgtactc tggttttacc tccgctaagg gtctatggac tgaacttgag 24840
aacgagttct ctgaagttga caatggcaat gagtcattca caacggaaaa ctaccttaac 24900
tataaaatgg ttgagggaag gtctgttatg gagcagctac aggagattca actccttgtt 24960
agggacttgg tccaatatgg ctgcgtccta ccagacagtt ttcaggtgaa tgcaatactg 25020
gcaaagctcc cgccttcttg gcgagacttt gtgacttcac gccgtcacat gaaaaagcaa 25080
atgactctca ctgagctgtc agctgcgata aatgtggaag agcgtgcaag gtccagtaac 25140
aagccatccc agcaactcca ggctcacgtt gttgagaagg gtggagatag aaaattccag 25200
aagaagaaga agaactctcc acagaagaat ctgaaccaac ctaaatccaa aaagatgaag 25260
aagaaaaagg aggactttat ctgctacgtt tgtggtgtct ccgggcatac agctcggagg 25320
tgcaaactta ggaagggaaa aggtcctcca cctcagcgca aagaagggaa cgtggtggtt 25380
aactccaccc cagggtatgc acttcaggcc tttatggcaa gtccatcaga tgactggtgg 25440
atggattccg gtgctactgt tcatatttgt gctgatcgat ccatgttctc ttcattccag 25500
ggatacaaca gcgcaccagt cttgatggga aatggtgttc cggcagcagt tcgaggtact 25560
ggacaggtct acttgaagtt aacttcagga aagacactcg tgctgaagga cgtactctat 25620
gtgccctcga tgagccggaa cctcatttca gtgtcgctgc tatgtcgaca gggtctcaaa 25680
ttagtgtttg agtctaataa agtggtcctg tctaagtttg gtacttttgt tggaaagtca 25740
tatgagtcag gcggtttgtt ccgtctttct gttttgaaca atcattcttc ttaccatgtt 25800
aatgtggtct gtaataacga ttccattaat aatatatggc attcccgttt gtgtcatgtg 25860
aattttgagg ccattaagag attaagtgac atgtctttaa ttcctgaata taaacatgtt 25920
aaaggtgtaa aatgtggtat ttgtgtgcaa gctaagcagc ctagaaaacc gtttcatacg 25980
gttgaaggca gaagtaccac tcctttagaa ctaattcact cagatatctg tgagatgaat 26040
ggtataatca caaaaggcgg taaaagatac tttcttacct tgatagatga tgctaccagg 26100
ttctgttata tttacttact cagaactaaa gatgaggcac tagaacactt taagatctat 26160
aagactgaag ttgaaaacca gttagacaag aaaattaaaa ggttacggtc tgatagagga 26220
ggtgaatacc tttccaacct ttttgatgag tactgcaaag agtgtggaat cattcatgaa 26280
accaccgctc catattcacc tcagtcaaat ggggtagctg aaaggaagaa ccgaacggtg 26340
tgtgacttag ctaatgcttt gttgcaaagt tcggggatgc ctgacatctg gtggggcgag 26400
gcagtactca cagtgtgcta tgtcctgaac agggtgccgc ctcgcaaccg tgaggccacg 26460
ccctatgaag gatttaaagg aaggaagcca gatctttcgc atcttcggac ttggggctgc 26520
cttgcaaagg tgaatgtccc gttgccgaag aagagaaagc taggccctaa gaccgtagac 26580
tgtgtcttcc tgggttatgc acataacagt gcagcttata gatttctagt ggtccattcc 26640
gagacaagcg aagtcgctgt aaatgtaata atggagtctc gggatgtgac attctttgag 26700
agcatcttcc ctatgcggga taaggaagta gtagccccca gatggtccat ctcggacata 26760
ttctctgccc tcgagtgtcc atgaccagac tccagatttg gagttaagga ggagtaagag 26820
acaaaggact gaaaagtctc tgggtgatga ttatattatt tatctagtgg atgaagaacc 26880
acgctccctc acagaggcat atacatctcc ggatgcagag tactggaggg aggctgtccg 26940
tagcgagatg gattcaatca tctcgaacgg aacttgggag ataactgatc tcccagctgg 27000
ttgcaagcct gtgggctgca aatggatctt taggaggaag aggagacccg atggtactat 27060
tgaaaagtac aaggcccgtc ttgtggctaa gggttttact caaaagaaag aggaggatta 27120
ttttgatact tattctccgg tggctcgatt gcccacaatc cgtgtgcttt tagcactggc 27180
agctgcttat aaacttcttg tccatcaaat ggacgtaaag acagcattcc taaatggaga 27240
gctggaagag gaaatttata tgcagcaacc agaaggattt gtggttaaag gtcaagagag 27300
caaagtgtgt cgcttgatta aatccttata cggtcttaag caagctccca gacaatggca 27360
tgagaagttt aataatactc tgaccactgc cgggttttgt gtaaacgaag ccgacaagtg 27420
tgtgtattat cgcttttctg ggggcaaagg tgtcatcatg tgtttatacg ttgatgacat 27480
attgatattt gggaccgatt tggaggctat catggagaca aaaatattcc tctccaagaa 27540
ctttgatatg aaggacttgg gtgaagctga tgtaatattg aacatcaagc taataaaggg 27600
tgaggatagg attactttgt cacaatccca ttatgtggaa aaggtcctga ctcgctttgg 27660
acatatggac tgtaaaccag tcaccacgcc ctatgatcca tcctacacgc tcagtaaata 27720
tgaaggcgag cctgtgaacc agctactata ttcgcagatc atcggatctc tcatgtacct 27780
gagcagtgca actagaccag atatctcata tgcagtatgc agactggctc gttattcggc 27840
cagtccagga gatagacact gggtagcctt gtacagagtg cttcggtatt tgaagggagc 27900
gatgaatctt ggtattaaat ataccggatt tccgtcagta cttgaaggat tcagtgatgc 27960
aaactggatc tctgactcgg atcaaatgaa gtctacaagt ggctatgtgt tcactttagc 28020
tggtggggcc gtgtcatgga gatcgtctaa acaatcagtg tcgacacgtt ccaccaagga 28080
ggctgaatta gttgcacttg attcagcagc attggaggct gaatggttga gagacctgtt 28140
gtcagacctt ccaatgctag caaagccgat accggctgtc ctagtatact gtgacaacac 28200
ttctgtgttg ctgaaagtga acagtagaaa ggacaaccaa aaatcctcga ggcatatcag 28260
aagacgactt gattcatgtc ggcatgctag agagacgggt gtaataaccg tagattacat 28320
caagtctgaa aggaaccttg ctgatccttt caccaaagga ctggctcaaa agccaatcca 28380
agcagcgtgc atgggaatgg gactcgtacc ctgttagcgg tttcaactgc ggataactgt 28440
cctgtgtgac cggagatccc gagatctagg ctccaggaaa cgaagcactg tgaaaccaag 28500
agcacaaaag ataaagagtc tcatgagctg ctgtgctctg tctgtatggc aggttgagcg 28560
atatgctctt aatgaagtca atgctataag cagggaaatt gtcctacaga atttccttaa 28620
cgatttcacc tatatgagct gactgtgggg tcgcagtcca ggagagaatg gggttttctc 28680
tctagtgagc tcatgaatag atattaaagg gcatgaccgt aacgccctgc cccgtaagag 28740
aagcagataa tctgcctagt actatgtgcc ttttgtgcga agattctcac actagggttt 28800
gtggatcaag gcgtagtcct ccgcttactc gtgcagggtt ggagtacact gggataggct 28860
ttggttcaaa cctaacaagt acctctgccg aaaagtagta tataaacagt acgggagctc 28920
aatgacattg atcagaaata agagtgaaaa tggtggggat tgttgtccat tttacgaatt 28980
ttcactttat taaaaaaaac aaaggggccg actgggtggg ccgctactag gctggaggcc 29040
gagcagccca ccggcaagcg ctggtacata accagctcgg gcacctgtgc gtcaaaaaag 29100
ggacagagag cggcggcgaa ggcacaactg cttggccggc ctgcacagcc gcctcttgcg 29160
tttctctctg ttcaacggcc tccgaatttg gtgcccggtc tgttccataa gtatgggtgt 29220
gtttcccctc tctctggcgt tggctgagca gtcggatgtt cttcttcctc cctctgtgtg 29280
tgttgctccg gtaagaactc aaactaccag ttcgtttcct ggtattgtgg tggcgattct 29340
tgctgggagt ggttcaaagt gggaggaagt ggtgctgtgc ttgcgcagtg tgagttacct 29400
gtgctctgtt gccggaatcg tgcacttctc cggtgctctg attcttgtct gatctctgtc 29460
caccgtggcc tggtgcttga gctccctgct gcgcggtgtt catcttctcg ggctggtgcc 29520
gtggacgtct ccctcggttc cggcagcggc tctagtatct ccgtcaacct cccgccgtgc 29580
aggcccggac gtgggcggcg gaattggttt tctgggacag aacttcgtgt tcgctgagct 29640
gatcttccca cgtagacaat ctacgtgttg tgagttattc gttgcctcac tttatttact 29700
gtttaatttt gtgcaatctg ctagtattaa atacttgggt gtgattgcac tgttgtggac 29760
ggatttgtgt gctcgatgat gatagcggtg tagtgagaca acgtgacagt ctgggtttta 29820
gtgtattatc attttcagca ttgatattat ttgtgcctaa ttaaatctct ctttgttaag 29880
gttgtttacc tgtgttaaat tttgttaatt tattaagtac acagctcctt tctatcagta 29940
tttatatttg attattaggc ttattacatc tgatctaaaa cccttatttg gtaagatagg 30000
actgtcacgt tagtcgagga tctctgatca cctattagcg ttggagggct gtttattaca 30060
gcaacctgct attttgagag caatatttta agataaacaa ttatgtttat ttgtttaatc 30120
tgccttcttc atctgcaagc catgtttaat ttccaaatgg acctgtgtag ctattttaca 30180
tgtttacatg tcttagaatt gctactgttc attgctgtta taattacata tggcatttgt 30240
ctgagttata tgcttgtttt attcggaatt agaatattta atttatttaa atatgaagga 30300
aacatgtcac ttatttctct atatttacaa cactaagttg tggtctgata caaatacaga 30360
acattaacat tatgtgaagc attaatgtga agcaagggga gagtaattac tccgttgagc 30420
caggttgaag agctctgtaa tatcaccctc acgttgctcg tcagccaaac ttggtacctg 30480
gaaacagcaa ttaccctcat aagcctctta aacgtaaacc aggaaagcaa ataaaaacag 30540
aaaataagtg gaattgtctg tggtgatgtt atccatgcct gatcgctatc tgcttcgacg 30600
tccaacctac aaaattccca gcaccccgac atatataaac atcatccgct ggtttaatcc 30660
aattcgagcg ctaataaaat cgagatgtag agggaaccca ccggacggaa ccggagctct 30720
cgcggcgggc cttaccatga gaatcgccgt tcccctccct gagacacgaa ttagaggacc 30780
gaaatcagct gaggaacagg cacgcataca gagtacagta ctacagttac acacccgcgc 30840
gcagcaatta tgggagcacg atcgcacgga gtggtgctgg gtgtgggatt tgcacgcacc 30900
tggaagagga gtaggaggag caccgggtta gaagcaccgg tggagcagcg cgggcggagg 30960
gggagcaatg tgccccgccg cacggacgca ggatacgccg tcgctgtgat cgaggagctg 31020
gtggcggggc ccgcgacgcc gcaccgacca cgaggcgcgg cgtctccatg gttgcctgct 31080
acgcacgctg ctcgtacgcc tccttggtta gctatgcgcg ccgtacctct ccgcggattg 31140
cccaggtgaa agagacaatt ccgtgtgctt caatggccag aggcgaggct gctgcctgat 31200
ggcggctggc gggcgctgag ccgctgcgct gatcggtgcg aggaggcgcc gttcggcgtg 31260
cgtgctcgcg gaggtgggac gtggcacgtg ggc 31293
<210> 2
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
cagacacgca ccaccatacg ctc 23
<210> 3
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
cagggacttg ggaacactcg gat 23
<210> 4
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
caggacgatg aagcttacgg g 21
<210> 5
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
taggaggctg aacgactgac ga 22
<210> 6
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
taaactccca ctgaaaatga a 21
<210> 7
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
ctgatgactt tgagcggtat 20
<210> 8
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
tggaataaac aacacaatta agact 25
<210> 9
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
gaagaagaac atccgactgc 20
<210> 10
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
ggagaaagcc aagtggg 17
<210> 11
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
tggagtcgtt attacagacc a 21
<210> 12
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
ggtcaagaga gcaaagtgtg tc 22
<210> 13
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
ccagcaagaa tcgccac 17
<210> 14
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
cacctcctgc ctgtgcgtca 20
<210> 15
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
ctgtatgcgt gcctgttcct ca 22
<210> 16
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
ttcccctccc tgagacacg 19
<210> 17
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
ctttcttggt tccactaacg ca 22
<210> 18
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 18
gagtcagcac ctaacccatc 20
<210> 19
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 19
gaacataata ccactacacc ataag 25
<210> 20
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 20
gttcccaagt ccctgaatgt 20
<210> 21
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
cacacacact aacgacagac g 21
<210> 22
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 22
caaagtcatc agggtagcca 20
<210> 23
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 23
gcaacctctc ttcactaacc g 21
<210> 24
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 24
cagtcttgat gggaaatggt g 21
<210> 25
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 25
cacacttgtc ggcttcgtt 19
<210> 26
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 26
gggtgaggat gggattactt t 21
<210> 27
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 27
ttgaaaccgc taacagggt 19
<210> 28
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 28
gagagacggg tgtaataacc g 21
<210> 29
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 29
ggcaacagag cacaggtaac 20
<210> 30
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 30
cgctaatgct acacgggat 19
<210> 31
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 31
cttatgctca gtaaaccgag actc 24
<210> 32
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 32
tggtttcgca taccgct 17
<210> 33
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 33
tgtgcctggt gactaacga 19
<210> 34
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 34
gctactggat agggatggta 20
<210> 35
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 35
attaaaggga gagataaagg t 21
<210> 36
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 36
aaaactgacc tggtgagcaa 20
<210> 37
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 37
aatcttattt cggtggcctt 20
<210> 38
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 38
ggtgtccaga cagtctatca ag 22
<210> 39
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 39
gaagctcatg tgtgttatcc a 21
<210> 40
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 40
gcatagaaca acagtcaaaa cc 22
<210> 41
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 41
gcttccttcc ttgctgataa a 21
<210> 42
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 42
aaggattagt tactaggctg g 21
<210> 43
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 43
gactgagaac atttgaacat tac 23
<210> 44
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 44
cagttcgttt cctggtattg t 21
<210> 45
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 45
tctttttgga tttaggttgg t 21
<210> 46
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 46
acgtgccacg tcccacctc 19
<210> 47
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 47
tcagatgctt ccgtcatgct ga 22
<210> 48
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 48
gatcacagcg acggcgtat 19
<210> 49
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 49
tcccaaagtt caccatccaa t 21
<210> 50
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 50
ataaactgga gatgtgtggg ct 22
<210> 51
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 51
aaaaaaacag gaaaaaagaa ggc 23
<210> 52
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 52
agtatggcga ggatttttgg 20
<210> 53
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 53
gcgatggtct cttcttgtgc 20
<210> 54
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 54
tcaacagcca ttccgcacta 20
<210> 55
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 55
ttttcccctt cctcccttct 20
<210> 56
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 56
ccctaatcat cctactgttg tcct 24
<210> 57
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 57
taatccgaag cactcgtcc 19
<210> 58
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 58
tcaacaaata taatatccct c 21
<210> 59
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 59
gatcaagtag tagtctccaa a 21
<210> 60
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 60
cagcccccac agaacagagc 20
<210> 61
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 61
gcaaagggag cgagcagaga 20
<210> 62
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 62
gtaaataagc aagggacagc ggt 23
<210> 63
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 63
agccaagcag gaggagtaag agt 23
<210> 64
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 64
ctccagcaag gaatccagta 20
<210> 65
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 65
gtaggaagac aaacagaaag tagtg 25
<210> 66
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 66
ctgctcttcc aatccaccac 20
<210> 67
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 67
cacgtctcca aacgtaaccc 20
<210> 68
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 68
aaacctcaaa ctgccacccc 20
<210> 69
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 69
ctctccttct ccaccgcctc 20
<210> 70
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 70
ccgccgtctc ttctttctt 19
<210> 71
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 71
ccagctcgtg cctccttat 19
<210> 72
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 72
atggagacgc cgcgcctcgt ggtcg 25
<210> 73
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 73
cgaccacgag gcgcggcgtc tccat 25
<210> 74
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 74
atggagagca gatggcggaa ggcca 25
<210> 75
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 75
tggccttccg ccatctgctc tccat 25

Claims (3)

1.一种调控玉米百粒重主效QTL定位，其特征在于，所述的玉米百粒重主效QTL为qKW4a，位于B73参考基因组4号染色体上，分子标记kw4和kw10之间。
2.一种如权利要求1所述的调控玉米百粒重主效QTL的候选基因，其特征在于，所述候选基因为ZmSSV，其基因序列为如下序列之一：
1)SEQ ID NO.1所示的序列；
2)在SEQ ID NO.1所示序列基础上发生一个或多个碱基的取代、缺失或替换且与SEQID NO.1所示的序列具有95％以上同一性的序列。
3.一种如权利要求1所述的调控玉米百粒重主效QTL或权利要求2所述的候选基因在玉米百粒重性状的遗传改良和分子育种中的应用。
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CN113637783A (zh) * 2021-06-01 2021-11-12 四川农业大学 基于线粒体基因组序列开发的老芒麦mtSSR标记引物及其应用

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GB9225845D0 (en) * 1992-12-10 1993-02-03 Nickerson Biocem Ltd Modified plants
CN104152450A (zh) * 2014-08-19 2014-11-19 上海交通大学 与黄瓜白粉病抗性主效基因共分离的InDel分子标记
CN109588308A (zh) * 2019-01-22 2019-04-09 重庆市农业科学院 一种vii型高直链淀粉玉米材料的选育方法

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GB9225845D0 (en) * 1992-12-10 1993-02-03 Nickerson Biocem Ltd Modified plants
CN104152450A (zh) * 2014-08-19 2014-11-19 上海交通大学 与黄瓜白粉病抗性主效基因共分离的InDel分子标记
CN109588308A (zh) * 2019-01-22 2019-04-09 重庆市农业科学院 一种vii型高直链淀粉玉米材料的选育方法

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113637783A (zh) * 2021-06-01 2021-11-12 四川农业大学 基于线粒体基因组序列开发的老芒麦mtSSR标记引物及其应用
CN113637783B (zh) * 2021-06-01 2023-04-25 四川农业大学 基于线粒体基因组序列开发的老芒麦mtSSR标记引物及其应用

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