CN111304214B - 一种增加香石竹花瓣数量的基因与应用 - Google Patents

一种增加香石竹花瓣数量的基因与应用 Download PDF

Info

Publication number
CN111304214B
CN111304214B CN202010098734.3A CN202010098734A CN111304214B CN 111304214 B CN111304214 B CN 111304214B CN 202010098734 A CN202010098734 A CN 202010098734A CN 111304214 B CN111304214 B CN 111304214B
Authority
CN
China
Prior art keywords
gene
carnation
petals
increasing
plant
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202010098734.3A
Other languages
English (en)
Other versions
CN111304214A (zh
Inventor
周旭红
瞿素萍
张颢
苏艳
田敏
李进昆
张露
罗柯佳
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Yunnan University of Traditional Chinese Medicine TCM
Flower Research Institute of YAAS
Original Assignee
Yunnan University of Traditional Chinese Medicine TCM
Flower Research Institute of YAAS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Yunnan University of Traditional Chinese Medicine TCM, Flower Research Institute of YAAS filed Critical Yunnan University of Traditional Chinese Medicine TCM
Priority to CN202010098734.3A priority Critical patent/CN111304214B/zh
Publication of CN111304214A publication Critical patent/CN111304214A/zh
Application granted granted Critical
Publication of CN111304214B publication Critical patent/CN111304214B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8262Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield involving plant development
    • C12N15/827Flower development or morphology, e.g. flowering promoting factor [FPF]

Abstract

本发明涉及一种增加香石竹花瓣数量的基因与应用,属于植物基因工程技术领域。本发明所述的增加香石竹花瓣数量的基因的RNA序列如SEQ ID NO.1所示。该基因命名为DcPS1。采用该基因增加香石竹花瓣数量的方法具体为:抑制所述DcPS1基因的基因表达量,得到花瓣增多的转基因植物。本发明首次在香石竹中发现了一个控制香石竹花瓣增多的基因,该基因控制花瓣数量的功能为植物中首次发现,为其他物种利用同源基因克隆相关基因提供了候选基因。本发明基因也可以为石竹科以及其他双子叶植物的花瓣增多的基因研究,提供依据。

Description

一种增加香石竹花瓣数量的基因与应用
技术领域
本发明属于植物基因工程技术领域,与植物转基因技术和香石竹分子育种技术领域相关,具体涉及一种增加香石竹花瓣数量的基因与应用。
背景技术
花卉是具有观赏价值的草本植物,花卉主要供观赏的部位有花色、花形、花香、花的大小、数目及花瓣数目等。花瓣数目是重要的观赏性状,培育重瓣性强的花卉新品种是观赏植物育种的重要目标。最早发现与花重瓣相关的基因是拟南芥中的C类基因AG基因,在ag突变体中,雄蕊和雌蕊被萼片和花瓣替代,并形成花中花的结构即重瓣花(Bowman JL,Smyth DR, Meyerowitz EM. Genes directing flower development inArabidopsis.Plant Cell, 1989, 1 (1): 37-52)。矮牵牛的C类基因PMADS3和FBP6两个基因的突变,植株均会出现柱头皱缩,花药瓣化,花朵呈现重瓣花(Heijmans K, Ament K,Rijpkema AS, Zethof J, Wolters-Arts M, Gerats T, Vandenbussche M.Redefining Cand Din the petunia ABC. Plant Cell, 2012, 24: 2305-2317)。
香石竹主要分布于欧洲温带以及中国的福建、湖北等地,原产于地中海地区。香石竹是肯尼亚也是美洲哥伦比亚主要的出口花卉品种,在亚洲的中国、日本、韩国、马来西亚等国都有大量栽培。在欧洲,德国、匈牙利、意大利、波兰、西班牙、土耳其、英国和荷兰等国栽培的规模都很大,是世界上应用最普遍的花卉之一。香石竹既可以作为盆栽植物来观赏,也可以当做切花的材料,同时在药用方面也有比较广泛的应用。在香石竹的研究中,重瓣花性状是不完全显性,重瓣花与单瓣花杂交后代中,花器官的表型有单瓣、重瓣和介于单瓣重瓣之间的类型(程金水,刘青林,园林植物遗传育种学.北京:中国林业出版社,2000,49-52)。有关控制香石竹花重瓣性的基因还未被发现,因此,有必要提供一种花卉花瓣控制基因,进而通过者转基因的方法培育一些重瓣的花卉新品种。
发明内容
本发明的目的是为了解决现有技术的不足,提供一种增加香石竹花瓣数量的基因与应用。
为实现上述目的,本发明采用的技术方案如下:
本发明第一方面提供一种增加香石竹花瓣数量的基因,所述的增加香石竹花瓣数量的基因的RNA序列如SEQ ID NO.1所示。该基因命名为DcPS1 (Dianthus CaryophyllusParallel Spindle 1)。
本发明第二方面提供含有所述的增加香石竹花瓣数量的基因的表达载体、表达盒、转基因细胞系或重组菌。
本发明第三方面提供所述的增加香石竹花瓣数量的基因在调控植物花瓣表型中的应用。
进一步,优选的是,所述的植物为石竹属植物。
本发明第四方面提供所述的增加香石竹花瓣数量的基因在植物育种中的应用。
进一步,优选的是,所述的植物为石竹属植物。
本发明第五方面提供一种增加香石竹花瓣数量的方法,包括如下步骤:
抑制所述增加香石竹花瓣数量的基因的基因表达量,得到花瓣增多的转基因植物。
本发明所述的抑制方法包括RNAi干扰技术和基因编缉技术,但不限于以上基因敲除方法。
本发明与现有技术相比,其有益效果为:
本发明首次在香石竹中发现了一个控制香石竹花瓣增多的基因,该基因控制花瓣数量的功能为植物中首次发现,为其他物种利用同源基因克隆相关基因提供了候选基因。
本发明中的基因也可以为石竹科以及其他双子叶植物的花瓣增多的基因研究提供依据。
附图说明
此外所说明的附图用来提供对本发明的进一步理解,构成本发明的一部分,本发明的示意性实施例及其说明用于解释本发明,并不构成对本发明的不当限定。
图1转基因植株和野生型DcPS1基因的表达量。
图2转基因植株和野生型花的对比图,转基因植株花瓣明显增多。
图3阳性转基因植株的PCR检测。注,+:阳性对照,含有pJL10-RNAI-FOR-REV载体质粒的PCR扩增产物;-:阴性对照,非转基因植株PCR扩增产物;H2O:水对照,以水为模板的PCR扩增产物;M:2000 Marker;1~11:待检测的植株。
具体实施方式
下面结合实施例对本发明作进一步的详细描述。
本领域技术人员将会理解,下列实施例仅用于说明本发明,而不应视为限定本发明的范围。实施例中未注明具体技术或条件者,按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用材料或设备未注明生产厂商者,均为可以通过购买获得的常规产品。
实施例1香石竹DcPS1基因RNAi表达载体构建
1)香石竹DcPS1基因RNAi干扰片断的PCR扩增
将DcPS1基因第1900-2399碱基500 bp长度的序列作为RNAi干扰片断进行PCR扩增:
正义干扰片断的PCR反应体系为:含有DcPS1基因的质粒作为模板,模板取10纳克,引物PJL10-RNAI-FOR-F(10微摩尔/升)和PJL10-RNAI-FOR-R(10微摩尔/升)各 2微升(表1),5×TransStart FastPfu Fly Buffer 10微升,2.5 毫摩尔/升 dNTPs 4微升,TransStartFastPfu Fly DNA Polymerase(北京全式金生物技术有限公司)1微升,双蒸水补充到50微升。
反义干扰片断的PCR反应体系为:含有DcPS1基因的质粒作为模板,模板取10纳克,引物PJL10-RNAI-REV-F(10微摩尔/升)和PJL10-RNAI-REV-R(10微摩尔/升) 2微升(表1),5×TransStart FastPfu Fly Buffer 10微升,2.5 毫摩尔/升 dNTPs 4微升,TransStartFastPfu Fly DNA Polymerase(北京全式金生物技术有限公司)1微升,双蒸水补充到50微升。
PCR反应条件:95℃ 5分钟;95℃ 30秒,55℃ 30秒,72℃ 3分钟,35个循环(从第2步到第4步);72℃ 5分钟。琼脂糖凝胶电泳,目的片段的回收,得到RNAI-FOR、RNAI-REV。
表1 引物的序列。FP:前引物,RP:后引物
Figure SMS_1
2) pJL10-RNAI-FOR载体构建
使用XhoI和KpnI核酸内切酶对pJL10载体酶切,所述的pJL10载体DNA序列如SEQID NO.14所示,并回收线性化的载体;使用pEASY Uni Seamless Cloning and AssembyKit (北京全式金生物技术有限公司)将RNAI-FOR(步骤1回收的正义干扰片断)拼接至线性化的pJL10载体上(pJL10载体为卡那抗性),并转化Trans2-Blue感受态细胞(北京全式金生物技术有限公司)。
阳性克隆的鉴定:对转化子进行菌液PCR鉴定,PCR反应体系:质粒10纳克,引物pJL10-FOR-iden-F(10 微摩尔/升)0.5微升,引物pJL10-FOR-iden-R (10微摩尔/升)0.5微升(表1),2×EasyTaq PCRSuperMix 10微升,双蒸水补充至20 微升。PCR反应条件:95 ℃ 5分钟;95 ℃ 30秒,55 ℃ 30秒,72 ℃ 3分钟,35个循环(从第2步到第4步);72 ℃ 5分钟。琼脂糖凝胶电泳观察结果。
测序:选取鉴定正确的3个阳性克隆送去Life Technologies公司测序,使用的测序引物为pJL10-FOR-iden-F和pJL10-FOR-iden-R。
3)pJL10-RNAI-FOR-REV载体构建:
选取测序结果与给定序列一致的pJL10-RNAI-FOR载体克隆进行后续实验。使用ClaI和XbaI核酸内切酶对pJL10-RNAI-FOR载体酶切,并回收线性化的载体;使用pEASY UniSeamless Cloning and Assemby Kit将RNAI-REV(步骤1回收的反义干扰片断)拼接至线性化的pJL10-RNAI-FOR载体上,并转化Trans2-Blue感受态细胞。
阳性克隆的鉴定:对转化子进行菌液PCR鉴定,PCR反应体系:质粒10纳克,引物pJL10-REV-iden-F(10 微摩尔/升)0.5微升,引物pJL10-REV-iden-R (10 微摩尔/升) 0.5微升(表1),2×EasyTaq PCRSuperMix 10微升,双蒸水补充至20微升。PCR反应条件: 95 ℃5分钟;95 ℃ 30秒,55 ℃ 30秒,72 ℃ 3分钟,35个循环(从第2步到第4步);72 ℃ 5分钟。琼脂糖凝胶电泳观察结果。
测序:选取鉴定正确的3个阳性克隆送去Life Technologies公司测序,使用的测序引物为pJL10-REV-iden-F和pJL10-REV-iden-R(表1)。
实施例2香石竹DcPS1基因遗传转化
1)植物表达载体转化农杆菌
采用冻融法将构建好的表达载体转入农杆菌,具体操作如下:向冰上刚融化的50微升农杆菌感受态细胞中加入1微克pJL10-RNAI-FOR-REV载体质粒DNA,混匀后冰上放置30分钟,液氮速冻5分钟,37 ℃水浴3分钟至全融,冰上放置2分钟,加入到1毫升 YEB液体培养基中,28 ℃,200 转/分恢复培养3小时,5000转/分离心3 分钟,收集菌体,均匀涂于事先加有50毫克/升Rif和50毫克/升 Kan的LB平板上,28 ℃倒置避光培养2天。
从转化平板上随机挑取10个单菌落,分别接种于50毫升含50毫克/升Rif和50毫克/升 Kan的YEB液体培养基中,28℃,200转/分恒温振荡培养2天,取少量菌液进行PCR扩增鉴定,PCR扩增的体系及程序为:通过PBI121-PRO-5309F和 PS1RNAI-982R 引物检测,引物的扩增体系为质粒50纳克,PBI121-PRO-5309F(10微摩尔/升)和PS1RNAI-982R(10微摩尔/升)引物(表1)各1微升,10× Taq Buffer 2微升,25毫摩尔/升 MgCl21.2微升,2.5毫摩尔/升dNTP 1.6微升,5U/微升 Taq 0.15微升,双蒸水补充至20微升。PCR反应条件:95 ℃ 5分钟;95 ℃ 30秒,56 ℃ 30秒,72 ℃ 1分钟,35个循环(从第2步到第4步);72 ℃ 5分钟。将鉴定的含目的载体阳性农杆菌与相同体积50%(体积百分比)的甘油混合均匀,置于-80 ℃冰箱内保存。
2)香石竹遗传转化体系
将-80 ℃保存的农杆菌接种到附加50毫克/升 Rif和50毫克/升Kan的YEB固体培养基平板上,将平板在28 ℃倒置培养2天,待菌落长出后,挑取单菌落接种到含50毫克/升Rif和50毫克/升 Kan的YEB液体培养基中,在28 ℃,200 转/分摇床上振荡培养过夜,得到预活化菌液;再取20微升预活化菌液接种于30毫升YEB液体培养基,继续在28 ℃,200转/分摇床上振荡,当OD600为0.5时,用于农杆菌浸染。
在超净工作台上,取无菌的香石竹组培苗放入体视显微镜下,用镊子小心的剥去顶端的叶片,露出茎尖;将茎尖切去生长点,然后将茎尖横切成5毫米的薄层,薄层背面朝下接种于MS+TDZ 0.5毫克/升+NAA 0.5毫克/升,蔗糖30克/升,琼脂7克/升,pH为5.8的预培养培养基中,24 ℃条件下光照预培养4天(光照强度1000-2000勒克斯),得到外植体。再浸入之前已活化备好的农杆菌菌液中30分钟,此过程放在摇床上,低速(30-50 rpm)不断振荡以使菌液与外植体充分接触。将外植体放在灭过菌的滤纸上吸干多余的菌液,转入共培养的培养基MS+TDZ 0.5 毫克/升+NAA 0.5毫克/升+AS 30毫克/升中在24 ℃条件下暗培养4天,再转入筛选培养基MS+TDZ 0.5毫克/升+NAA 0.1毫克/升+Cef 400毫克/升 +kan 50毫克/升中进行筛选培养,60天转接一次,待分化出再生芽,将再生芽转入MS+BA 0.1毫克/升+NAA0.1毫克/升+Kan 50毫克/升增殖培养基中进行增殖筛选培养。
3)香石竹阳性转基因植株的验证
pJL10-RNAI-FOR-REV载体阳性转基因植株采用PCR法验证,首先提取的转基因植株DNA,通过PJL10-50F和PJL10-469R引物检测,引物序列如表1所示。引物均扩出含目的条带的片断为RNAi载体阳性转基因再生植株(图3),图3中共检测11株植株,其中,编号分别为5、9、10、11的植株为阳性转基因植株
实施例3香石竹转基因阳性植株DcPS1表达和表型
采用实时定量荧光PCR,对DcPS1基因的表达进行分析,发现DcPS1基因在转基因阳性植株中的表达量下降(图1),通过构建DcPS1基因RNAi载体获得转基因植株‘170’并进行组培及扦插扩繁,与野生型相比花瓣数由平均29枚增多为36枚,花瓣数明显增多(图2)。
为了证明DcPS1为多花瓣控制的基因,我们发现另三株阳性转基因植株出现与上述阳性转基因植株相同的表型。本发明公开的内容可为研究DcPS1基因在植株中调控花瓣数量打下基础。
以上显示和描述了本发明的基本原理、主要特征和本发明的优点。本行业的技术人员应该了解,本发明不受上述实施例的限制,上述实施例和说明书中描述的只是说明本发明的原理,在不脱离本发明精神和范围的前提下,本发明还会有各种变化和改进,这些变化和改进都落入要求保护的本发明范围内。本发明要求保护范围由所附的权利要求书及其等效物界定。
序列表
<110> 云南省农业科学院花卉研究所
云南中医药大学
<120> 一种增加香石竹花瓣数量的基因与应用
<160> 14
<170> SIPOSequenceListing 1.0
<210> 1
<211> 3423
<212> RNA
<213> 人工序列
<400> 1
augucgaaac uccaaaaaga gagagaaauc ccaguuuuca caguuuggaagaacaaugcc 60
auacucaaga acaucuuccu caucgauaau ccuccuaauu ucaacgauaa ucaagaacgu 120
aacgacgaug uaauucaacg agaagucgaa gaaacguugu uagucggucg acauccagau 180
ugugauauuc gaguggaaca uccgaguauu agcagauuuc aucuccgcau ucauucguua240
ccgacauuga aucuccucuuuauuacugau uuaucuucug ugcacgggac guggguguca 300
gauaaaagga uugaaccgaa cauaagaacg ccgauaaaag ugggugauca ucuuaggauu360
ggagggucua guaggguuua caagcuucac uggauuccuu ugagccuugc uuaugacgug420
ucgaaaccau uugugccgcc ccuugauuuc ucugugcagc gcgaagauga ggacgaaauc480
gucaaggacg agauuucagu aggguuagaa gacgagaagg ugaauaaugu cgauaacauu540
ucugaugacg uuucaaacgu ucuggagucu uuggaucuau guaaugguga ugagacgguu600
acucagaaac cuguuuuguu ggaagaugug gguacugaau ugguggaugu caauuucgaa660
cauugcgaug gcauaucucg agaacuugug cgaaaagaaa ugcuccugga accucuuacu720
ggagacaaug aaucugaaaa cgaagauauu cagucaaugg acucucuucu ggaaggguug780
gugccucuuu ucuuggguga gaacugggaa uggguugguc aggaugagaa cugucuucca840
cguucgacag aagacgauaa ucaucugcug ucuucuaaag aaaccgaaaa agaucuuggc900
ggugcaguau uggagggauc aaaugauucg uuuucaauug agaauuuaga acuuuuaacu960
cagagagaga ccucgucagc gacucuugua cccgaacaca uggaauuucc uaaggcagaa1020
aacguggaug acaucaaagc ugaaggaaga caggauccua cagaaguaga ucaucgucua1080
ccgucaacug gauuuguuag cccgguguug gagggaucaa auguuacguu uguggaugag1140
aacucugaau uguuacacca gaaaaccacc ucauuggaag cuccaauuuc agaagacgug1200
gaauuuucua cggcagaaaa cauggauggc aucacagcug aaggaagaca ggauccuaca1260
gaaguagauc auugucuacc gucaacagga uuuguuagcu cagugaugga gggaucaaau1320
guuacguuug uggaugagaa cucuggauug uuacaccaga aaaccaccuc acuggaagcu1380
ccaauuucag aagaugugaa auuuucuacg gcagaaaaca uggauggcau cacagcugau1440
ggaguacaga aggaauuagg aucugaaguu uuaggagaag ccguugaucc auggaguuuc1500
ugguguggaa cacuuguaga aucaucauca ucuccuguca cugcagagua ucagccacuu1560
gucgauacuu uauuccuaga cgagaacuuc ggacugcuag cucagaaagg cacaucauca1620
gcaacagaaa acauauauaa ccucauugcu gaagaaaaaa acaauaaaau uguugaagaa1680
gauguugaag auaaagccug gaguuuuugg ugugggaguc aucaaccugg uguagaaaau1740
gacaagacaa caaacgaauc agcugaagca uugucuaaaa aagcauccuc aggaagcucg1800
aguucagacc ugaauucacc gaccaaggaa cacccaggaa augaacugug gaguuuuugg1860
ucaggaaaga ugggggucga auccguuugc ucagcauugu ccgaugcaga acccuucuca1920
gauucggaau auaaaucguu gccaaauggu uaucugaguu ccccuaugau aucgucagga1980
caaaacucug cccacgguuu uugguccaag agagaauuac cgaaagauac aucuugugga2040
gacuguuauc gaagcccaac auccaacagu ucugucauag ggcgauuggg gaguccucuu2100
augacaucug accagaaccc uauggauagu auuugguuga gaagagguaa auuagcgagc2160
gcuccucaag uucuaaccag cagaaccgaa gaaaagaaag gaaacucaga ucagaagaac2220
gccaagcgug aauccaucuc gaggaugcuu uuuaaaggga augaugauga ggaggaggag2280
gaagaggaag aagucuucac ucccgauaaa gaaaauuuca ccccuacuac acacaagucc2340
augagaaagu uuacaauuuu gaaagagauu aauaauaauu cgaguccagg caaaucaccc2400
cuuuccaaga ugacuuuuag uccugacagc cguugcgugg augucgucuc ucccuuguca2460
gauaaagaaa auuacucgac aagaacuaaa ggaaagaaga aaacugcaac acgugguucc2520
gcaaaucgag uacgaucagu caagcagaaa gagagaaaug caguucggau gccauuccaa2580
ucgcugcuag cuaauucacc ccagaagagu gauacugaau acuacauugc ugagcuugaa2640
acaagaaaug cgaguuccgu cgaugaucca aauguuauca cacgaaaggu aaaaugcguu2700
aaagaggaac uucaacagug gaacauggua guggauacga caacucuacu gaacaaggaa2760
ucgagaaagg cauugcagcu ucuugagggu cugaaaggaa ccaaacuaau cguccccaga2820
auagugauca cggagcuuuc aagcuugaaa cgacaguuaa gcuucuucag aagaaauacg2880
gagguacugu cagccuugga auggauacaa gaauguaugc ugaaaacugg augguggauu2940
cagguccgaa acucggcgga agaaggccga ccaguugcac cgacaccucc cgcuucaccu3000
caaacugccu cauacucguu cucaggcuuu ucaucacgaa cagaaguucu uucacccacu3060
guugaagacg auguacucga auuugcucuc ggcuucagga agaguaaccc ugauggacaa3120
cucguucucc ucaguaauga caucucuaug aagaucaaag cuaugacaga ggguaugauc3180
ugugagacug gagaagaguu cagagaaagu uuggugaacc cguuuuccga gagauucagg3240
uggacugaca gcuccccucg aggucaaacu uggucuugug cagacgacuu uguuuugaag3300
gagagauuuu acccaagcuc acugaacaug augaccucaa aguccagagg aacuguaaag3360
gguuugaagc ugauacuuca acacaauucu caguaucaug aacgagugaa cgcagcagcc3420
uaa3423
<210>2
<211>46
<212>DNA
<213>人工序列
<400>2
ttcatttgga gaggacacgc tcgagtccga tgcagaacccttctca 46
<210> 3
<211> 46
<212> DNA
<213> 人工序列
<400> 3
ttatttcctt accaattggg gtaccggtga tttgcctggactcgaa 46
<210> 4
<211> 46
<212> DNA
<213> 人工序列
<400> 4
ttgggaaatt gggttcgaaa tcgatggtga tttgcctggactcgaa 46
<210> 5
<211> 46
<212> DNA
<213> 人工序列
<400> 5
tatctcatta aagcaggact ctagatccga tgcagaacccttctca 46
<210> 6
<211> 20
<212> DNA
<213> 人工序列
<400> 6
tggaaaaaga agacgttcca 20
<210> 7
<211> 20
<212> DNA
<213> 人工序列
<400> 7
cttactctca acttttatct 20
<210> 8
<211> 20
<212> DNA
<213> 人工序列
<400> 8
cttgtagttt tattaacttc 20
<210> 9
<211> 20
<212> DNA
<213> 人工序列
<400> 9
gtaaggatct gagctacaca 20
<210> 10
<211> 20
<212> DNA
<213> 人工序列
<400> 10
acagaactcg ccgtaaagac 20
<210> 11
<211> 20
<212> DNA
<213> 人工序列
<400> 11
aggaggagga agaggaagaa 20
<210> 12
<211> 20
<212> DNA
<213> 人工序列
<400> 12
cactatcctt cgcaagaccc 20
<210> 13
<211> 20
<212> DNA
<213> 人工序列
<400> 13
cctcctcatc atcattccct 20
<210> 14
<211> 14792
<212> DNA
<213> 人工序列
<400> 14
ggggtggaag gtggagaagg agtccagaga ctggttaatg gtcgaggccacgaccgtcag 60
acgacagcgg tgactgctcg acgtggtgat ggtgatgacg gtcgagggaa caggggtcag120
ggtaccccaa ttggtaagga aataattatt ttcttttttc cttttagtat aaaatagtta180
agtgatgtta attagtatga ttataataat atagttgtta taattgtgaa aaaataattt240
ataaatatat tgtttacata aacaacatag taatgtaaaa aaatatgaca agtgatgtgt300
aagacgaaga agataaaagt tgagagtaag tatattattt ttaatgaatt tgatcgaaca360
tgtaagatga tatactagca ttaatatttg ttttaatcat aatagtaatt ctagctggtt420
tgatgaatta aatatcaatg ataaaatact atagtaaaaa taagaataaa taaattaaaa480
taatattttt ttatgattaa tagtttatta tataattaaa tatctatacc attactaaat540
attttagttt aaaagttaat aaatattttg ttagaaattc caatctgctt gtaatttatc600
aataaacaaa atattaaata acaagctaaa gtaacaaata atatcaaact aatagaaaca660
gtaatctaat gtaacaaaac ataatctaat gctaatataa caaagcgcaa gatctatcat720
tttatatagt attattttca atcaacattc ttattaattt ctaaataata cttgtagttt780
tattaacttc taaatggatt gactattaat taaatgaatt agtcgaacat gaataaacaa840
ggtaacatga tagatcatgt cattgtgtta tcattgatct tacatttgga ttgattacag900
ttgggaaatt gggttcgaaa tcgatctgac ccctgttccc tcgaccgtca tcaccatcac960
cacgtcgagc agtcaccgct gtcgtctgac ggtcgtggcc tcgaccatta accagtctct1020
ggactccttc tccaccttcc acccctctag agtcctgctt taatgagata tgcgagacgc1080
ctatgatcgc atgatatttg ctttcaattc tgttgtgcac gttgtaaaaa acctgagcat1140
gtgtagctca gatccttacc gccggtttcg gttcattcta atgaatatat cacccgttac1200
tatcgtattt ttatgaataa tattctccgt tcaatttact gattgtaccc tactacttat1260
atgtacaata ttaaaatgaa aacaatatat tgtgctgaat aggtttatag cgacatctat1320
gatagagcgc cacaataaca aacaattgcg ttttattatt acaaatccaa ttttaaaaaa1380
agcggcagaa ccggtcaaac ctaaaagact gattacataa atcttattca aatttcaaaa1440
ggccccaggg gctagtatct acgacacacc gagcggcgaa ctaataacgt tcactgaagg1500
gaactccggt tccccgccgg cgcgcatggg tgagattcct tgaagttgag tattggccgt1560
ccgctctacc gaaagttacg ggcaccattc aacccggtcc agcacggcgg ccgggtaacc1620
gacttgctgc cccgagaatt atgcagcatt tttttggtgt atgtgggccc caaatgaagt1680
gcaggtcaaa ccttgacagt gacgacaaat cgttgggcgg gtccagggcg aattttgcga1740
caacatgtcg aggctcagca ggacctgcag gcatgcaagc tagcttacta gtgatatccc1800
gcggccatgg cggccgggag catgcgacgt cgggcccaat tcgccctata gtgagtcgta1860
ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca1920
acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg1980
caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgtaaacgtt2040
aatgggtttc tggagtttaa tgagctaagc acatacgtca gaaaccatta ttgcgcgttc2100
aaaagtcgcc taaggtcact atcagctagc aaatatttct tgtcaaaaat gctccactga2160
cgttccataa attcccctcg gtatccaatt agagtctcat attcactctc aatccaaata2220
atctgcaatg gcaattacct tatccgcaac ttctttacctatttccgccc ggatccgggc2280
aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat2340
cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt2400
caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg2460
gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag2520
ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc2580
tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc2640
tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga2700
agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga2760
actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg2820
cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg2880
tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc2940
tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc3000
cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg3060
gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc3120
gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc3180
ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccgatccaac acttacgttt3240
gcaacgtcca agagcaaata gaccacgaac gccggaaggt tgccgcagcg tgtggattgc3300
gtctcaattc tctcttgcag gaatgcaatg atgaatatga tactgactat gaaactttga3360
gggaatactg cctagcaccg tcacctcata acgtgcatca tgcatgccct gacaacatgg3420
aacatcgcta tttttctgaa gaattatgct cgttggagga tgtcgcggca attgcagcta3480
ttgccaacat cgaactaccc ctcacgcatg cattcatcaa tattattcat gcggggaaag3540
gcaagattaa tccaactggc aaatcatcca gcgtgattgg taacttcagt tccagcgact3600
tgattcgttt tggtgctacc cacgttttca ataaggacga gatggtggag taaagaagga3660
gtgcgtcgaa gcagatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt3720
tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat3780
taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt3840
atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg3900
cgcggtgtca tctatgttac tagatcgaat taattccagg cggtgaaggg caatcagctg3960
ttgcccgtct cactggtgaa aagaaaaacc accccagtac attaaaaacg tccgcaatgt4020
gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag4080
ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc4140
agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc4200
gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg4260
agggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatca aatatcatct4320
ccctcgcaga gatccgaatt atcagccttc ttattcattt ctcgcttaac cgtgacaggc4380
tgtcgatctt gagaactatg ccgacataat aggaaatcgc tggataaagc cgctgaggaa4440
gctgagtggc gctatttctt tagaagtgaa cgttgacgat gtcgacggat cttttccgct4500
gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca4560
cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt4620
cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc4680
ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccggctacc4740
gccggcgtaa cagatgaggg caagcggatg gctgatgaaa ccaagccaac caggggtgat4800
gctgccaact tactgattta gtgtatgatg gtgtttttga ggtgctccag tggcttctgt4860
ttctatcagc tgtccctcct gttcagctac tgacggggtg gtgcgtaacg gcaaaagcac4920
cgccggacat cagcgctatc tctgctctca ctgccgtaaa acatggcaac tgcagttcac4980
ttacaccgct tctcaacccg gtacgcacca gaaaatcatt gatatggcca tgaatggcgt5040
tggatgccgg gcaacagccc gcattatggg cgttggcctc aacacgattt tacgtcactt5100
aaaaaactca ggccgcagtc ggtaacctcg cgcatacagc cgggcagtga cgtcatcgtc5160
tgcgcggaaa tggacgaaca gtggggctat gtcggggcta aatcgcgcca gcgctggctg5220
ttttacgcgt atgacagtct ccggaagacg gttgttgcgc acgtattcgg tgaacgcact5280
atggcgacgc tggggcgtct tatgagcctg ctgtcaccct ttgacgtggt gatatggatg5340
acggatggct ggccgctgta tgaatcccgc ctgaagggaa agctgcacgt aatcagcaag5400
cgatatacgc agcgaattga gcggcataac ctgaatctga ggcagcacct ggcacggctg5460
ggacggaagt cgctgtcgtt ctcaaaatcg gtggagctgc atgacaaagt catcgggcat5520
tatctgaaca taaaacacta tcaataagtt ggagtcatta cccaaccagg aagggcagcc5580
cacctatcaa ggtgtactgc cttccagacg aacgaagagc gattgaggaa aaggcggcgg5640
cggccggcat gagcctgtcg gcctacctgc tggccgtcgg ccagggctac aaaatcacgg5700
gcgtcgtgga ctatgagcac gtccgcgagc tggcccgcat caatggcgac ctgggccgcc5760
tgggcggcct gctgaaactc tggctcaccg acgacccgcg cacggcgcgg ttcggtgatg5820
ccacgatcct cgccctgctg gcgaagatcg aagagaagca ggacgagctt ggcaaggtca5880
tgatgggcgt ggtccgcccg agggcagagc catgactttt ttagccgcta aaacggccgg5940
ggggtgcgcg tgattgccaa gcacgtcccc atgcgctcca tcaagaagag cgacttcgcg6000
gagctggtat tcgtgcaggg caagattcgg aataccaagt acgagaagga cggccagacg6060
gtctacggga ccgacttcat tgccgataag gtggattatc tggacaccaa ggcaccaggc6120
gggtcaaatc aggaataagg gcacattgcc ccggcgtgag tcggggcaat cccgcaagga6180
gggtgaatga atcggacgtt tgaccggaag gcatacaggc aagaactgat cgacgcgggg6240
ttttccgccg aggatgccga aaccatcgca agccgcaccg tcatgcgtgc gccccgcgaa6300
accttccagt ccgtcggctc gatggtccag caagctacgg ccaagatcga gcgcgacagc6360
gtgcaactgg ctccccctgc cctgcccgcg ccatcggccg ccgtggagcg ttcgcgtcgt6420
ctcgaacagg aggcggcagg tttggcgaag tcgatgacca tcgacacgcg aggaactatg6480
acgaccaaga agcgaaaaac cgccggcgag gacctggcaa aacaggtcag cgaggccaag6540
caggccgcgt tgctgaaaca cacgaagcag cagatcaagg aaatgcagct ttccttgttc6600
gatattgcgc cgtggccgga cacgatgcga gcgatgccaa acgacacggc ccgctctgcc6660
ctgttcacca cgcgcaacaa gaaaatcccg cgcgaggcgc tgcaaaacaa ggtcattttc6720
cacgtcaaca aggacgtgaa gatcacctac accggcgtcg agctgcgggc cgacgatgac6780
gaactggtgt ggcagcaggt gttggagtac gcgaagcgca cccctatcgg cgagccgatc6840
accttcacgt tctacgagct ttgccaggac ctgggctggt cgatcaatgg ccggtattac6900
acgaaggccg aggaatgcct gtcgcgccta caggcgacgg cgatgggctt cacgtccgac6960
cgcgttgggc acctggaatc ggtgtcgctg ctgcaccgct tccgcgtcct ggaccgtggc7020
aagaaaacgt cccgttgcca ggtcctgatc gacgaggaaa tcgtcgtgct gtttgctggc7080
gaccactaca cgaaattcat atgggagaag taccgcaagc tgtcgccgac ggcccgacgg7140
atgttcgact atttcagctc gcaccgggag ccgtacccgc tcaagctgga aaccttccgc7200
ctcatgtgcg gatcggattc cacccgcgtg aagaagtggc gcgagcaggt cggcgaagcc7260
tgcgaagagt tgcgaggcag cggcctggtg gaacacgcct gggtcaatga tgacctggtg7320
cattgcaaac gctagggcct tgtggggtca gttccggctg ggggttcagc agccagcgct7380
ttactggcat ttcaggaaca agcgggcact gctcgacgca cttgcttcgc tcagtatcgc7440
tcgggacgca cggcgcgctc tacgaactgc cgataaacag aggattaaaa ttgacaattg7500
tgattaaggc tcagattcga cggcttggag cggccgacgt gcaggatttc cgcgagatcc7560
gattgtcggc cctgaagaaa gctccagaga tgttcgggtc cgtttacgag cacgaggaga7620
aaaagcccat ggaggcgttc gctgaacggt tgcgagatgc cgtggcattc ggcgcctaca7680
tcgacggcga gatcattggg ctgtcggtct tcaaacagga ggacggcccc aaggacgctc7740
acaaggcgca tctgtccggc gttttcgtgg agcccgaaca gcgaggccga ggggtcgccg7800
gtatgctgct gcgggcgttg ccggcgggtt tattgctcgt gatgatcgtc cgacagattc7860
caacgggaat ctggtggatg cgcatcttca tcctcggcgc acttaatatt tcgctattct7920
ggagcttgtt gtttatttcg gtctaccgcc tgccgggcgg ggtcgcggcg acggtaggcg7980
ctgtgcagcc gctgatggtc gtgttcatct ctgccgctct gctaggtagc ccgatacgat8040
tgatggcggt cctgggggct atttgcggaa ctgcgggcgt ggcgctgttg gtgttgacac8100
caaacgcagc gctagatcct gtcggcgtcg cagcgggcct ggcgggggcg gtttccatgg8160
cgttcggaac cgtgctgacc cgcaagtggc aacctcccgt gcctctgctc acctttaccg8220
cctggcaact ggcggccgga ggacttctgc tcgttccagt agctttagtg tttgatccgc8280
caatcccgat gcctacagga accaatgttc tcggcctggc gtggctcggc ctgatcggag8340
cgggtttaac ctacttcctt tggttccggg ggatctcgcg actcgaacct acagttgttt8400
ccttactggg ctttctcagc cgggatggcg ctaagaagct attgccgccg atcttcatat8460
gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc8520
ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca8580
ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg8640
agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca8700
taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa8760
cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc8820
tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc8880
gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct8940
gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg9000
tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag9060
gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta9120
cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg9180
aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt9240
tgtttgcaag cagcagatta cgcgcagaaa aaaaggatat caagaagatc ctttgatctt9300
ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag9360
attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat9420
ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc9480
tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat9540
aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc9600
acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag9660
aagtggtcct gcaactttat ccgcctccat ccagtctatt aaacaagtgg cagcaacgga9720
ttcgcaaacc tgtcacgcct tttgtgccaa aagccgcgcc aggtttgcga tccgctgtgc9780
caggcgttag gcgtcatatg aagatttcgg tgatccctga gcaggtggcg gaaacattgg9840
atgctgagaa ccatttcatt gttcgtgaag tgttcgatgt gcacctatcc gaccaaggct9900
ttgaactatc taccagaagt gtgagcccct accggaagga ttacatctcg gatgatgact9960
ctgatgaaga ctctgcttgc tatggcgcat tcatcgacca agagcttgtc gggaagattg 10020
aactcaactc aacatggaac gatctagcct ctatcgaaca cattgttgtg tcgcacacgc 10080
accgaggcaa aggagtcgcg cacagtctca tcgaatttgc gaaaaagtgg gcactaagca 10140
gacagctcct tggcatacga ttagagacac aaacgaacaa tgtacctgcc tgcaatttgt 10200
acgcaaaatg tggctttact ctcggcggca ttgacctgtt cacgtataaa actagacctc 10260
aagtctcgaa cgaaacagcg atgtactggt actggttctc gggagcacag gatgacgcct 10320
aacaattcat tcaagccgac accgcttcgc ggcgcggctt aattcaggag ttaaacatca 10380
tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg 10440
agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg 10500
gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa 10560
caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg 10620
agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt 10680
atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta 10740
tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac 10800
atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg 10860
atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg 10920
gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca 10980
aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc 11040
agcccgtcat acttgaagct aggcaggctt atcttggaca agaagatcgc ttggcctcgc 11100
gcgcagatca gttggaagaa tttgttcact acgtgaaagg cgagatcacc aaggtagtcg 11160
gcaaataatg tctaacaatt cgttcaagcc gacgccgctt cgcggcgcgg cttaactcaa 11220
gcgttagaga gctggggaag actatgcgcg atctgttgaa ggtggttcta agcctcgtac 11280
ttgcgatggc atcggggcag gcacttgctg acctgccaat tgttttagtg gatgaagctc 11340
gtcttcccta tgactactcc ccatccaact acgacatttc tccaagcaac tacgacaact 11400
ccataagcaa ttacgacaat agtccatcaa attacgacaa ctctgagagc aactacgata 11460
atagttcatc caattacgac aatagtcgca acggaaatcg taggcttata tatagcgcaa 11520
atgggtctcg cactttcgcc ggctactacg tcattgccaa caatgggaca acgaacttct 11580
tttccacatc tggcaaaagg atgttctaca ccccaaaagg ggggcgcggc gtctatggcg 11640
gcaaagatgg gagcttctgc ggggcattgg tcgtcataaa tggccaattt tcgcttgccc 11700
tgacagataa cggcctgaag atcatgtatc taagcaacta gcctgctctc taataaaatg 11760
ttaggagctt ggctgccatt tttggggtga ggccgttcgc ggccgagggg cgcagcccct 11820
ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg ggcacccccc 11880
ttcggcgtgc gcggtcacgc gccagggcgc agccctggtt aaaaacaagg tttataaata 11940
ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg cggaaaccct 12000
tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc 12060
atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg 12120
cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac atcatctgtg 12180
ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag ctccacgtcg 12240
ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag tcggcccctc 12300
aagtgtcaac gtccgcccct catctgtcag tgagggccaa gttttccgcg aggtatccac 12360
aacgccggcg gccggccgcg gtgtctcgca cacggcttcg acggcgtttc tggcgcgttt 12420
gcagggccat agacggccgc cagcccagcg gcgagggcaa ccagcccggt gagcgtcgga 12480
aagggtcgac atcttgctgc gttcggatat tttcgtggag ttcccgccac agacccggat 12540
tgaaggcgag atccagcaac tcgcgccaga tcatcctgtg acggaacttt ggcgcgtgat 12600
gactggccag gacgtcggcc gaaagagcga caagcagatc acgattttcg acagcgtcgg 12660
atttgcgatc gaggattttt cggcgctgcg ctacgtccgc gaccgcgttg agggatcaag 12720
ccacagcagc ccactcgacc ttctagccga cccagacgag ccaagggatc tttttggaat 12780
gctgctccgt cgtcaggctt tccgacgttt gggtggttga acagaagtca ttatcgtacg 12840
gaatgccagc actcccgagg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg 12900
gataaacctt ttcacgccct tttaaatatc cgttattcta ataaacgctc ttttctctta 12960
ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt aaactgaagg cgggaaacga 13020
caatctgatc atgagcggag aattaaggga gtcacgttat gacccccgcc gatgacgcgg 13080
gacaagccgt tttacgtttg gaactgacag aaccgcaacg attgaaggag ccactcagcc 13140
ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac 13200
aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact 13260
cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg 13320
agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctatttagg 13380
tgacactata gaatactcaa gctatgcatc caacgcgttg ggagctcgtc gagcggccgc 13440
tcgacgaatt aattccaatc ccacaaaaat ctgagcttaa cagcacagtt gctcctctca 13500
gagcagaatc gggtattcaa caccctcata tcaactacta cgttgtgtat aacggtccac 13560
atgccggtat atacgatgac tggggttgta caaaggcggc aacaaacggc gttcccggag 13620
ttgcacacaa gaaatttgcc actattacag aggcaagagc agcagctgac gcgtacacaa 13680
caagtcagca aacagacagg ttgaacttca tccccaaagg agaagctcaa ctcaagccca 13740
agagctttgc taaggcccta acaagcccac caaagcaaaa agcccactgg ctcacgctag 13800
gaaccaaaag gcccagcagt gatccagccc caaaagagat ctcctttgcc ccggagatta 13860
caatggacga tttcctctat ctttacgatc taggaaggaa gttcgaaggt gaaggtgacg 13920
acactatgtt caccactgat aatgagaagg ttagcctctt caatttcaga aagaatgctg 13980
acccacagat ggttagagag gcctacgcag caggtctcat caagacgatc tacccgagta 14040
acaatctcca ggagatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca 14100
ggactaattg catcaagaac acagagaaag acatatttct caagatcaga agtactattc 14160
cagtatggac gattcaaggc ttgcttcata aaccaaggca agtaatagag attggagtct 14220
ctaaaaaggt agttcctact gaatctaagg ccatgcatgg agtctaagat tcaaatcgag 14280
gatctaacag aactcgccgt gaagactggc gaacagttca tacagagtct tttacgactc 14340
aatgacaaga agaaaatctt cgtcaacatg gtggagcacg acactctggt ctactccaaa 14400
aatgtcaaag atacagtctc agaagaccaa agggctattg agacttttca acaaaggata 14460
atttcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat cgaaaggaca 14520
gtagaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggctatcatt 14580
caagatctct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg 14640
gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga catctccact 14700
gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga 14760
agttcatttcatttggagaggacacgctcgag 14792。
序列表
<110> 云南省农业科学院花卉研究所
云南中医药大学
<120> 一种增加香石竹花瓣数量的基因与应用
<160> 14
<170> SIPOSequenceListing 1.0
<210> 1
<211> 3423
<212> RNA
<213> 人工序列()
<400> 1
augucgaaac uccaaaaaga gagagaaauc ccaguuuuca caguuuggaa gaacaaugcc 60
auacucaaga acaucuuccu caucgauaau ccuccuaauu ucaacgauaa ucaagaacgu 120
aacgacgaug uaauucaacg agaagucgaa gaaacguugu uagucggucg acauccagau 180
ugugauauuc gaguggaaca uccgaguauu agcagauuuc aucuccgcau ucauucguua 240
ccgacauuga aucuccucuu uauuacugau uuaucuucug ugcacgggac guggguguca 300
gauaaaagga uugaaccgaa cauaagaacg ccgauaaaag ugggugauca ucuuaggauu 360
ggagggucua guaggguuua caagcuucac uggauuccuu ugagccuugc uuaugacgug 420
ucgaaaccau uugugccgcc ccuugauuuc ucugugcagc gcgaagauga ggacgaaauc 480
gucaaggacg agauuucagu aggguuagaa gacgagaagg ugaauaaugu cgauaacauu 540
ucugaugacg uuucaaacgu ucuggagucu uuggaucuau guaaugguga ugagacgguu 600
acucagaaac cuguuuuguu ggaagaugug gguacugaau ugguggaugu caauuucgaa 660
cauugcgaug gcauaucucg agaacuugug cgaaaagaaa ugcuccugga accucuuacu 720
ggagacaaug aaucugaaaa cgaagauauu cagucaaugg acucucuucu ggaaggguug 780
gugccucuuu ucuuggguga gaacugggaa uggguugguc aggaugagaa cugucuucca 840
cguucgacag aagacgauaa ucaucugcug ucuucuaaag aaaccgaaaa agaucuuggc 900
ggugcaguau uggagggauc aaaugauucg uuuucaauug agaauuuaga acuuuuaacu 960
cagagagaga ccucgucagc gacucuugua cccgaacaca uggaauuucc uaaggcagaa 1020
aacguggaug acaucaaagc ugaaggaaga caggauccua cagaaguaga ucaucgucua 1080
ccgucaacug gauuuguuag cccgguguug gagggaucaa auguuacguu uguggaugag 1140
aacucugaau uguuacacca gaaaaccacc ucauuggaag cuccaauuuc agaagacgug 1200
gaauuuucua cggcagaaaa cauggauggc aucacagcug aaggaagaca ggauccuaca 1260
gaaguagauc auugucuacc gucaacagga uuuguuagcu cagugaugga gggaucaaau 1320
guuacguuug uggaugagaa cucuggauug uuacaccaga aaaccaccuc acuggaagcu 1380
ccaauuucag aagaugugaa auuuucuacg gcagaaaaca uggauggcau cacagcugau 1440
ggaguacaga aggaauuagg aucugaaguu uuaggagaag ccguugaucc auggaguuuc 1500
ugguguggaa cacuuguaga aucaucauca ucuccuguca cugcagagua ucagccacuu 1560
gucgauacuu uauuccuaga cgagaacuuc ggacugcuag cucagaaagg cacaucauca 1620
gcaacagaaa acauauauaa ccucauugcu gaagaaaaaa acaauaaaau uguugaagaa 1680
gauguugaag auaaagccug gaguuuuugg ugugggaguc aucaaccugg uguagaaaau 1740
gacaagacaa caaacgaauc agcugaagca uugucuaaaa aagcauccuc aggaagcucg 1800
aguucagacc ugaauucacc gaccaaggaa cacccaggaa augaacugug gaguuuuugg 1860
ucaggaaaga ugggggucga auccguuugc ucagcauugu ccgaugcaga acccuucuca 1920
gauucggaau auaaaucguu gccaaauggu uaucugaguu ccccuaugau aucgucagga 1980
caaaacucug cccacgguuu uugguccaag agagaauuac cgaaagauac aucuugugga 2040
gacuguuauc gaagcccaac auccaacagu ucugucauag ggcgauuggg gaguccucuu 2100
augacaucug accagaaccc uauggauagu auuugguuga gaagagguaa auuagcgagc 2160
gcuccucaag uucuaaccag cagaaccgaa gaaaagaaag gaaacucaga ucagaagaac 2220
gccaagcgug aauccaucuc gaggaugcuu uuuaaaggga augaugauga ggaggaggag 2280
gaagaggaag aagucuucac ucccgauaaa gaaaauuuca ccccuacuac acacaagucc 2340
augagaaagu uuacaauuuu gaaagagauu aauaauaauu cgaguccagg caaaucaccc 2400
cuuuccaaga ugacuuuuag uccugacagc cguugcgugg augucgucuc ucccuuguca 2460
gauaaagaaa auuacucgac aagaacuaaa ggaaagaaga aaacugcaac acgugguucc 2520
gcaaaucgag uacgaucagu caagcagaaa gagagaaaug caguucggau gccauuccaa 2580
ucgcugcuag cuaauucacc ccagaagagu gauacugaau acuacauugc ugagcuugaa 2640
acaagaaaug cgaguuccgu cgaugaucca aauguuauca cacgaaaggu aaaaugcguu 2700
aaagaggaac uucaacagug gaacauggua guggauacga caacucuacu gaacaaggaa 2760
ucgagaaagg cauugcagcu ucuugagggu cugaaaggaa ccaaacuaau cguccccaga 2820
auagugauca cggagcuuuc aagcuugaaa cgacaguuaa gcuucuucag aagaaauacg 2880
gagguacugu cagccuugga auggauacaa gaauguaugc ugaaaacugg augguggauu 2940
cagguccgaa acucggcgga agaaggccga ccaguugcac cgacaccucc cgcuucaccu 3000
caaacugccu cauacucguu cucaggcuuu ucaucacgaa cagaaguucu uucacccacu 3060
guugaagacg auguacucga auuugcucuc ggcuucagga agaguaaccc ugauggacaa 3120
cucguucucc ucaguaauga caucucuaug aagaucaaag cuaugacaga ggguaugauc 3180
ugugagacug gagaagaguu cagagaaagu uuggugaacc cguuuuccga gagauucagg 3240
uggacugaca gcuccccucg aggucaaacu uggucuugug cagacgacuu uguuuugaag 3300
gagagauuuu acccaagcuc acugaacaug augaccucaa aguccagagg aacuguaaag 3360
gguuugaagc ugauacuuca acacaauucu caguaucaug aacgagugaa cgcagcagcc 3420
uaa 3423
<210> 2
<211> 46
<212> DNA
<213> 人工序列()
<400> 2
ttcatttgga gaggacacgc tcgagtccga tgcagaaccc ttctca 46
<210> 3
<211> 46
<212> DNA
<213> 人工序列()
<400> 3
ttatttcctt accaattggg gtaccggtga tttgcctgga ctcgaa 46
<210> 4
<211> 46
<212> DNA
<213> 人工序列()
<400> 4
ttgggaaatt gggttcgaaa tcgatggtga tttgcctgga ctcgaa 46
<210> 5
<211> 46
<212> DNA
<213> 人工序列()
<400> 5
tatctcatta aagcaggact ctagatccga tgcagaaccc ttctca 46
<210> 6
<211> 20
<212> DNA
<213> 人工序列()
<400> 6
tggaaaaaga agacgttcca 20
<210> 7
<211> 20
<212> DNA
<213> 人工序列()
<400> 7
cttactctca acttttatct 20
<210> 8
<211> 20
<212> DNA
<213> 人工序列()
<400> 8
cttgtagttt tattaacttc 20
<210> 9
<211> 20
<212> DNA
<213> 人工序列()
<400> 9
gtaaggatct gagctacaca 20
<210> 10
<211> 20
<212> DNA
<213> 人工序列()
<400> 10
acagaactcg ccgtaaagac 20
<210> 11
<211> 20
<212> DNA
<213> 人工序列()
<400> 11
aggaggagga agaggaagaa 20
<210> 12
<211> 20
<212> DNA
<213> 人工序列()
<400> 12
cactatcctt cgcaagaccc 20
<210> 13
<211> 20
<212> DNA
<213> 人工序列()
<400> 13
cctcctcatc atcattccct 20
<210> 14
<211> 14792
<212> DNA
<213> 人工序列()
<400> 14
ggggtggaag gtggagaagg agtccagaga ctggttaatg gtcgaggcca cgaccgtcag 60
acgacagcgg tgactgctcg acgtggtgat ggtgatgacg gtcgagggaa caggggtcag 120
ggtaccccaa ttggtaagga aataattatt ttcttttttc cttttagtat aaaatagtta 180
agtgatgtta attagtatga ttataataat atagttgtta taattgtgaa aaaataattt 240
ataaatatat tgtttacata aacaacatag taatgtaaaa aaatatgaca agtgatgtgt 300
aagacgaaga agataaaagt tgagagtaag tatattattt ttaatgaatt tgatcgaaca 360
tgtaagatga tatactagca ttaatatttg ttttaatcat aatagtaatt ctagctggtt 420
tgatgaatta aatatcaatg ataaaatact atagtaaaaa taagaataaa taaattaaaa 480
taatattttt ttatgattaa tagtttatta tataattaaa tatctatacc attactaaat 540
attttagttt aaaagttaat aaatattttg ttagaaattc caatctgctt gtaatttatc 600
aataaacaaa atattaaata acaagctaaa gtaacaaata atatcaaact aatagaaaca 660
gtaatctaat gtaacaaaac ataatctaat gctaatataa caaagcgcaa gatctatcat 720
tttatatagt attattttca atcaacattc ttattaattt ctaaataata cttgtagttt 780
tattaacttc taaatggatt gactattaat taaatgaatt agtcgaacat gaataaacaa 840
ggtaacatga tagatcatgt cattgtgtta tcattgatct tacatttgga ttgattacag 900
ttgggaaatt gggttcgaaa tcgatctgac ccctgttccc tcgaccgtca tcaccatcac 960
cacgtcgagc agtcaccgct gtcgtctgac ggtcgtggcc tcgaccatta accagtctct 1020
ggactccttc tccaccttcc acccctctag agtcctgctt taatgagata tgcgagacgc 1080
ctatgatcgc atgatatttg ctttcaattc tgttgtgcac gttgtaaaaa acctgagcat 1140
gtgtagctca gatccttacc gccggtttcg gttcattcta atgaatatat cacccgttac 1200
tatcgtattt ttatgaataa tattctccgt tcaatttact gattgtaccc tactacttat 1260
atgtacaata ttaaaatgaa aacaatatat tgtgctgaat aggtttatag cgacatctat 1320
gatagagcgc cacaataaca aacaattgcg ttttattatt acaaatccaa ttttaaaaaa 1380
agcggcagaa ccggtcaaac ctaaaagact gattacataa atcttattca aatttcaaaa 1440
ggccccaggg gctagtatct acgacacacc gagcggcgaa ctaataacgt tcactgaagg 1500
gaactccggt tccccgccgg cgcgcatggg tgagattcct tgaagttgag tattggccgt 1560
ccgctctacc gaaagttacg ggcaccattc aacccggtcc agcacggcgg ccgggtaacc 1620
gacttgctgc cccgagaatt atgcagcatt tttttggtgt atgtgggccc caaatgaagt 1680
gcaggtcaaa ccttgacagt gacgacaaat cgttgggcgg gtccagggcg aattttgcga 1740
caacatgtcg aggctcagca ggacctgcag gcatgcaagc tagcttacta gtgatatccc 1800
gcggccatgg cggccgggag catgcgacgt cgggcccaat tcgccctata gtgagtcgta 1860
ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca 1920
acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg 1980
caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgtaaacgtt 2040
aatgggtttc tggagtttaa tgagctaagc acatacgtca gaaaccatta ttgcgcgttc 2100
aaaagtcgcc taaggtcact atcagctagc aaatatttct tgtcaaaaat gctccactga 2160
cgttccataa attcccctcg gtatccaatt agagtctcat attcactctc aatccaaata 2220
atctgcaatg gcaattacct tatccgcaac ttctttacct atttccgccc ggatccgggc 2280
aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 2340
cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 2400
caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg 2460
gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 2520
ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 2580
tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 2640
tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 2700
agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 2760
actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg 2820
cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 2880
tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 2940
tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 3000
cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 3060
gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 3120
gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 3180
ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccgatccaac acttacgttt 3240
gcaacgtcca agagcaaata gaccacgaac gccggaaggt tgccgcagcg tgtggattgc 3300
gtctcaattc tctcttgcag gaatgcaatg atgaatatga tactgactat gaaactttga 3360
gggaatactg cctagcaccg tcacctcata acgtgcatca tgcatgccct gacaacatgg 3420
aacatcgcta tttttctgaa gaattatgct cgttggagga tgtcgcggca attgcagcta 3480
ttgccaacat cgaactaccc ctcacgcatg cattcatcaa tattattcat gcggggaaag 3540
gcaagattaa tccaactggc aaatcatcca gcgtgattgg taacttcagt tccagcgact 3600
tgattcgttt tggtgctacc cacgttttca ataaggacga gatggtggag taaagaagga 3660
gtgcgtcgaa gcagatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt 3720
tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat 3780
taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt 3840
atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg 3900
cgcggtgtca tctatgttac tagatcgaat taattccagg cggtgaaggg caatcagctg 3960
ttgcccgtct cactggtgaa aagaaaaacc accccagtac attaaaaacg tccgcaatgt 4020
gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag 4080
ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc 4140
agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc 4200
gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg 4260
agggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatca aatatcatct 4320
ccctcgcaga gatccgaatt atcagccttc ttattcattt ctcgcttaac cgtgacaggc 4380
tgtcgatctt gagaactatg ccgacataat aggaaatcgc tggataaagc cgctgaggaa 4440
gctgagtggc gctatttctt tagaagtgaa cgttgacgat gtcgacggat cttttccgct 4500
gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca 4560
cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt 4620
cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc 4680
ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccggctacc 4740
gccggcgtaa cagatgaggg caagcggatg gctgatgaaa ccaagccaac caggggtgat 4800
gctgccaact tactgattta gtgtatgatg gtgtttttga ggtgctccag tggcttctgt 4860
ttctatcagc tgtccctcct gttcagctac tgacggggtg gtgcgtaacg gcaaaagcac 4920
cgccggacat cagcgctatc tctgctctca ctgccgtaaa acatggcaac tgcagttcac 4980
ttacaccgct tctcaacccg gtacgcacca gaaaatcatt gatatggcca tgaatggcgt 5040
tggatgccgg gcaacagccc gcattatggg cgttggcctc aacacgattt tacgtcactt 5100
aaaaaactca ggccgcagtc ggtaacctcg cgcatacagc cgggcagtga cgtcatcgtc 5160
tgcgcggaaa tggacgaaca gtggggctat gtcggggcta aatcgcgcca gcgctggctg 5220
ttttacgcgt atgacagtct ccggaagacg gttgttgcgc acgtattcgg tgaacgcact 5280
atggcgacgc tggggcgtct tatgagcctg ctgtcaccct ttgacgtggt gatatggatg 5340
acggatggct ggccgctgta tgaatcccgc ctgaagggaa agctgcacgt aatcagcaag 5400
cgatatacgc agcgaattga gcggcataac ctgaatctga ggcagcacct ggcacggctg 5460
ggacggaagt cgctgtcgtt ctcaaaatcg gtggagctgc atgacaaagt catcgggcat 5520
tatctgaaca taaaacacta tcaataagtt ggagtcatta cccaaccagg aagggcagcc 5580
cacctatcaa ggtgtactgc cttccagacg aacgaagagc gattgaggaa aaggcggcgg 5640
cggccggcat gagcctgtcg gcctacctgc tggccgtcgg ccagggctac aaaatcacgg 5700
gcgtcgtgga ctatgagcac gtccgcgagc tggcccgcat caatggcgac ctgggccgcc 5760
tgggcggcct gctgaaactc tggctcaccg acgacccgcg cacggcgcgg ttcggtgatg 5820
ccacgatcct cgccctgctg gcgaagatcg aagagaagca ggacgagctt ggcaaggtca 5880
tgatgggcgt ggtccgcccg agggcagagc catgactttt ttagccgcta aaacggccgg 5940
ggggtgcgcg tgattgccaa gcacgtcccc atgcgctcca tcaagaagag cgacttcgcg 6000
gagctggtat tcgtgcaggg caagattcgg aataccaagt acgagaagga cggccagacg 6060
gtctacggga ccgacttcat tgccgataag gtggattatc tggacaccaa ggcaccaggc 6120
gggtcaaatc aggaataagg gcacattgcc ccggcgtgag tcggggcaat cccgcaagga 6180
gggtgaatga atcggacgtt tgaccggaag gcatacaggc aagaactgat cgacgcgggg 6240
ttttccgccg aggatgccga aaccatcgca agccgcaccg tcatgcgtgc gccccgcgaa 6300
accttccagt ccgtcggctc gatggtccag caagctacgg ccaagatcga gcgcgacagc 6360
gtgcaactgg ctccccctgc cctgcccgcg ccatcggccg ccgtggagcg ttcgcgtcgt 6420
ctcgaacagg aggcggcagg tttggcgaag tcgatgacca tcgacacgcg aggaactatg 6480
acgaccaaga agcgaaaaac cgccggcgag gacctggcaa aacaggtcag cgaggccaag 6540
caggccgcgt tgctgaaaca cacgaagcag cagatcaagg aaatgcagct ttccttgttc 6600
gatattgcgc cgtggccgga cacgatgcga gcgatgccaa acgacacggc ccgctctgcc 6660
ctgttcacca cgcgcaacaa gaaaatcccg cgcgaggcgc tgcaaaacaa ggtcattttc 6720
cacgtcaaca aggacgtgaa gatcacctac accggcgtcg agctgcgggc cgacgatgac 6780
gaactggtgt ggcagcaggt gttggagtac gcgaagcgca cccctatcgg cgagccgatc 6840
accttcacgt tctacgagct ttgccaggac ctgggctggt cgatcaatgg ccggtattac 6900
acgaaggccg aggaatgcct gtcgcgccta caggcgacgg cgatgggctt cacgtccgac 6960
cgcgttgggc acctggaatc ggtgtcgctg ctgcaccgct tccgcgtcct ggaccgtggc 7020
aagaaaacgt cccgttgcca ggtcctgatc gacgaggaaa tcgtcgtgct gtttgctggc 7080
gaccactaca cgaaattcat atgggagaag taccgcaagc tgtcgccgac ggcccgacgg 7140
atgttcgact atttcagctc gcaccgggag ccgtacccgc tcaagctgga aaccttccgc 7200
ctcatgtgcg gatcggattc cacccgcgtg aagaagtggc gcgagcaggt cggcgaagcc 7260
tgcgaagagt tgcgaggcag cggcctggtg gaacacgcct gggtcaatga tgacctggtg 7320
cattgcaaac gctagggcct tgtggggtca gttccggctg ggggttcagc agccagcgct 7380
ttactggcat ttcaggaaca agcgggcact gctcgacgca cttgcttcgc tcagtatcgc 7440
tcgggacgca cggcgcgctc tacgaactgc cgataaacag aggattaaaa ttgacaattg 7500
tgattaaggc tcagattcga cggcttggag cggccgacgt gcaggatttc cgcgagatcc 7560
gattgtcggc cctgaagaaa gctccagaga tgttcgggtc cgtttacgag cacgaggaga 7620
aaaagcccat ggaggcgttc gctgaacggt tgcgagatgc cgtggcattc ggcgcctaca 7680
tcgacggcga gatcattggg ctgtcggtct tcaaacagga ggacggcccc aaggacgctc 7740
acaaggcgca tctgtccggc gttttcgtgg agcccgaaca gcgaggccga ggggtcgccg 7800
gtatgctgct gcgggcgttg ccggcgggtt tattgctcgt gatgatcgtc cgacagattc 7860
caacgggaat ctggtggatg cgcatcttca tcctcggcgc acttaatatt tcgctattct 7920
ggagcttgtt gtttatttcg gtctaccgcc tgccgggcgg ggtcgcggcg acggtaggcg 7980
ctgtgcagcc gctgatggtc gtgttcatct ctgccgctct gctaggtagc ccgatacgat 8040
tgatggcggt cctgggggct atttgcggaa ctgcgggcgt ggcgctgttg gtgttgacac 8100
caaacgcagc gctagatcct gtcggcgtcg cagcgggcct ggcgggggcg gtttccatgg 8160
cgttcggaac cgtgctgacc cgcaagtggc aacctcccgt gcctctgctc acctttaccg 8220
cctggcaact ggcggccgga ggacttctgc tcgttccagt agctttagtg tttgatccgc 8280
caatcccgat gcctacagga accaatgttc tcggcctggc gtggctcggc ctgatcggag 8340
cgggtttaac ctacttcctt tggttccggg ggatctcgcg actcgaacct acagttgttt 8400
ccttactggg ctttctcagc cgggatggcg ctaagaagct attgccgccg atcttcatat 8460
gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc 8520
ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 8580
ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 8640
agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 8700
taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 8760
cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 8820
tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 8880
gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 8940
gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 9000
tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 9060
gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 9120
cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 9180
aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 9240
tgtttgcaag cagcagatta cgcgcagaaa aaaaggatat caagaagatc ctttgatctt 9300
ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 9360
attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 9420
ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 9480
tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 9540
aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 9600
acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 9660
aagtggtcct gcaactttat ccgcctccat ccagtctatt aaacaagtgg cagcaacgga 9720
ttcgcaaacc tgtcacgcct tttgtgccaa aagccgcgcc aggtttgcga tccgctgtgc 9780
caggcgttag gcgtcatatg aagatttcgg tgatccctga gcaggtggcg gaaacattgg 9840
atgctgagaa ccatttcatt gttcgtgaag tgttcgatgt gcacctatcc gaccaaggct 9900
ttgaactatc taccagaagt gtgagcccct accggaagga ttacatctcg gatgatgact 9960
ctgatgaaga ctctgcttgc tatggcgcat tcatcgacca agagcttgtc gggaagattg 10020
aactcaactc aacatggaac gatctagcct ctatcgaaca cattgttgtg tcgcacacgc 10080
accgaggcaa aggagtcgcg cacagtctca tcgaatttgc gaaaaagtgg gcactaagca 10140
gacagctcct tggcatacga ttagagacac aaacgaacaa tgtacctgcc tgcaatttgt 10200
acgcaaaatg tggctttact ctcggcggca ttgacctgtt cacgtataaa actagacctc 10260
aagtctcgaa cgaaacagcg atgtactggt actggttctc gggagcacag gatgacgcct 10320
aacaattcat tcaagccgac accgcttcgc ggcgcggctt aattcaggag ttaaacatca 10380
tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg 10440
agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg 10500
gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa 10560
caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg 10620
agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt 10680
atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta 10740
tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac 10800
atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg 10860
atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg 10920
gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca 10980
aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc 11040
agcccgtcat acttgaagct aggcaggctt atcttggaca agaagatcgc ttggcctcgc 11100
gcgcagatca gttggaagaa tttgttcact acgtgaaagg cgagatcacc aaggtagtcg 11160
gcaaataatg tctaacaatt cgttcaagcc gacgccgctt cgcggcgcgg cttaactcaa 11220
gcgttagaga gctggggaag actatgcgcg atctgttgaa ggtggttcta agcctcgtac 11280
ttgcgatggc atcggggcag gcacttgctg acctgccaat tgttttagtg gatgaagctc 11340
gtcttcccta tgactactcc ccatccaact acgacatttc tccaagcaac tacgacaact 11400
ccataagcaa ttacgacaat agtccatcaa attacgacaa ctctgagagc aactacgata 11460
atagttcatc caattacgac aatagtcgca acggaaatcg taggcttata tatagcgcaa 11520
atgggtctcg cactttcgcc ggctactacg tcattgccaa caatgggaca acgaacttct 11580
tttccacatc tggcaaaagg atgttctaca ccccaaaagg ggggcgcggc gtctatggcg 11640
gcaaagatgg gagcttctgc ggggcattgg tcgtcataaa tggccaattt tcgcttgccc 11700
tgacagataa cggcctgaag atcatgtatc taagcaacta gcctgctctc taataaaatg 11760
ttaggagctt ggctgccatt tttggggtga ggccgttcgc ggccgagggg cgcagcccct 11820
ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg ggcacccccc 11880
ttcggcgtgc gcggtcacgc gccagggcgc agccctggtt aaaaacaagg tttataaata 11940
ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg cggaaaccct 12000
tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc 12060
atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg 12120
cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac atcatctgtg 12180
ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag ctccacgtcg 12240
ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag tcggcccctc 12300
aagtgtcaac gtccgcccct catctgtcag tgagggccaa gttttccgcg aggtatccac 12360
aacgccggcg gccggccgcg gtgtctcgca cacggcttcg acggcgtttc tggcgcgttt 12420
gcagggccat agacggccgc cagcccagcg gcgagggcaa ccagcccggt gagcgtcgga 12480
aagggtcgac atcttgctgc gttcggatat tttcgtggag ttcccgccac agacccggat 12540
tgaaggcgag atccagcaac tcgcgccaga tcatcctgtg acggaacttt ggcgcgtgat 12600
gactggccag gacgtcggcc gaaagagcga caagcagatc acgattttcg acagcgtcgg 12660
atttgcgatc gaggattttt cggcgctgcg ctacgtccgc gaccgcgttg agggatcaag 12720
ccacagcagc ccactcgacc ttctagccga cccagacgag ccaagggatc tttttggaat 12780
gctgctccgt cgtcaggctt tccgacgttt gggtggttga acagaagtca ttatcgtacg 12840
gaatgccagc actcccgagg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg 12900
gataaacctt ttcacgccct tttaaatatc cgttattcta ataaacgctc ttttctctta 12960
ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt aaactgaagg cgggaaacga 13020
caatctgatc atgagcggag aattaaggga gtcacgttat gacccccgcc gatgacgcgg 13080
gacaagccgt tttacgtttg gaactgacag aaccgcaacg attgaaggag ccactcagcc 13140
ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac 13200
aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact 13260
cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg 13320
agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctatttagg 13380
tgacactata gaatactcaa gctatgcatc caacgcgttg ggagctcgtc gagcggccgc 13440
tcgacgaatt aattccaatc ccacaaaaat ctgagcttaa cagcacagtt gctcctctca 13500
gagcagaatc gggtattcaa caccctcata tcaactacta cgttgtgtat aacggtccac 13560
atgccggtat atacgatgac tggggttgta caaaggcggc aacaaacggc gttcccggag 13620
ttgcacacaa gaaatttgcc actattacag aggcaagagc agcagctgac gcgtacacaa 13680
caagtcagca aacagacagg ttgaacttca tccccaaagg agaagctcaa ctcaagccca 13740
agagctttgc taaggcccta acaagcccac caaagcaaaa agcccactgg ctcacgctag 13800
gaaccaaaag gcccagcagt gatccagccc caaaagagat ctcctttgcc ccggagatta 13860
caatggacga tttcctctat ctttacgatc taggaaggaa gttcgaaggt gaaggtgacg 13920
acactatgtt caccactgat aatgagaagg ttagcctctt caatttcaga aagaatgctg 13980
acccacagat ggttagagag gcctacgcag caggtctcat caagacgatc tacccgagta 14040
acaatctcca ggagatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca 14100
ggactaattg catcaagaac acagagaaag acatatttct caagatcaga agtactattc 14160
cagtatggac gattcaaggc ttgcttcata aaccaaggca agtaatagag attggagtct 14220
ctaaaaaggt agttcctact gaatctaagg ccatgcatgg agtctaagat tcaaatcgag 14280
gatctaacag aactcgccgt gaagactggc gaacagttca tacagagtct tttacgactc 14340
aatgacaaga agaaaatctt cgtcaacatg gtggagcacg acactctggt ctactccaaa 14400
aatgtcaaag atacagtctc agaagaccaa agggctattg agacttttca acaaaggata 14460
atttcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat cgaaaggaca 14520
gtagaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggctatcatt 14580
caagatctct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg 14640
gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga catctccact 14700
gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga 14760
agttcatttc atttggagag gacacgctcg ag 14792

Claims (3)

1.一种增加香石竹花瓣数量的基因在调控石竹属植物花瓣数量中的应用,其特征在于,所述增加香石竹花瓣数量的基因的RNA序列如SEQ ID NO.1所示。
2.一种增加香石竹花瓣数量的基因在石竹属植物育种中的应用,其特征在于,所述增加香石竹花瓣数量的基因的RNA序列如SEQ ID NO.1所示。
3.一种增加香石竹花瓣数量的方法,其特征在于,包括如下步骤:
抑制权利要求1所述增加香石竹花瓣数量的基因的基因表达量,得到花瓣增多的转基因植物。
CN202010098734.3A 2020-02-18 2020-02-18 一种增加香石竹花瓣数量的基因与应用 Active CN111304214B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010098734.3A CN111304214B (zh) 2020-02-18 2020-02-18 一种增加香石竹花瓣数量的基因与应用

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010098734.3A CN111304214B (zh) 2020-02-18 2020-02-18 一种增加香石竹花瓣数量的基因与应用

Publications (2)

Publication Number Publication Date
CN111304214A CN111304214A (zh) 2020-06-19
CN111304214B true CN111304214B (zh) 2023-05-30

Family

ID=71154824

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010098734.3A Active CN111304214B (zh) 2020-02-18 2020-02-18 一种增加香石竹花瓣数量的基因与应用

Country Status (1)

Country Link
CN (1) CN111304214B (zh)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010061181A1 (en) * 2008-11-28 2010-06-03 The University Of Birmingham Engineering of plants to exhibit self-incompatibility
CN103756997A (zh) * 2014-01-18 2014-04-30 云南云科花卉有限公司 香石竹ps1基因5`侧翼未知序列的克隆方法
CN104195142A (zh) * 2014-08-29 2014-12-10 云南省农业科学院花卉研究所 一种香石竹PS1基因ihpRNA载体的构建方法
CN105838733A (zh) * 2016-05-18 2016-08-10 云南省农业科学院花卉研究所 Cas9 介导的香石竹基因编辑载体和应用
CN111593057A (zh) * 2020-05-12 2020-08-28 云南中医药大学 一种增加香石竹花朵直径的基因与应用

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010061181A1 (en) * 2008-11-28 2010-06-03 The University Of Birmingham Engineering of plants to exhibit self-incompatibility
CN103756997A (zh) * 2014-01-18 2014-04-30 云南云科花卉有限公司 香石竹ps1基因5`侧翼未知序列的克隆方法
CN104195142A (zh) * 2014-08-29 2014-12-10 云南省农业科学院花卉研究所 一种香石竹PS1基因ihpRNA载体的构建方法
CN105838733A (zh) * 2016-05-18 2016-08-10 云南省农业科学院花卉研究所 Cas9 介导的香石竹基因编辑载体和应用
CN111593057A (zh) * 2020-05-12 2020-08-28 云南中医药大学 一种增加香石竹花朵直径的基因与应用

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
Cytological, molecular mechanisms and temperature stress regulating production of diploid male gametes in Dianthus caryophyllus L;Zhou X等;《Plant Physiol Biochem》;20151022;第97卷;255-63 *
Mutations in AtPS1 (Arabidopsis thaliana parallel spindle 1) lead to the production of diploid pollen grains;d"Erfurth I等;《PLoS Genet》;20081128;第4卷(第11期);e1000274 *
The DcPS1 cooperates with OSDLa during pollen development and 2n gamete production in carnation meiosis;Zhou X等;《BMC Plant Biol》;20220524;第22卷(第1期);259 *
切花菊和香石竹采后主要衰老生理特性研究;张燕琴;《中国学位论文全文数据库》;20101018;全文 *
国家自然科学基金委员会生命科学部2017年度青年科学基金项目;2017年度青年科学基金项目;《生命科学》;20171215(第12期);92-157 *
环境因素对香石竹2n配子形成的影响及其形成机制研究;周旭红;《中国博士学位论文全文数据库农业科技辑》;20170115(第01期);D048-21 *
香石竹DcSKP1基因克隆及表达分析;周旭红等;《广西植物》;20220228;第42卷(第2期);286-293 *
香石竹GA20-oxidase基因的克隆及RNA干扰载体的构建;金亮等;《农业生物技术学报》;20090215(第01期);111-117 *

Also Published As

Publication number Publication date
CN111304214A (zh) 2020-06-19

Similar Documents

Publication Publication Date Title
CA2478910C (en) Methods and means for monitoring and modulating gene silencing
US20030049835A1 (en) Methods and means for producing efficient silencing construct using recombinational cloning
CN109722439B (zh) 烟草mlo2、mlo6和mlo12基因在制备抗白粉菌烟草品种中的应用及其方法
CN106939316B (zh) 利用CRISPR/Cas9系统定点敲除水稻OsPDCD5基因第二外显子的方法
CN110387384B (zh) 一种根癌农杆菌介导的桃熏草莓遗传转化方法
CN110724685A (zh) 转基因耐盐耐除草剂玉米sr801外源插入旁侧序列及其应用
AU2005252598B8 (en) Transformation vectors
KR20220091473A (ko) 유전자 변형 식물 및 이를 제조하는 방법
CN111593057B (zh) 一种增加香石竹花朵直径的基因与应用
CN109355306A (zh) 陆地棉转化事件icr24-397及其特异性鉴定方法
Díaz et al. Induction of hairy roots for symbiotic gene expression studies
CN111304214B (zh) 一种增加香石竹花瓣数量的基因与应用
AU759037B2 (en) Method for the induction of pathogen resistance in plants
CN114438104B (zh) 一种调控番茄果实糖含量的SlGRAS9基因及在培育高糖含量番茄中的应用
KR20220091472A (ko) 유전자 변형 식물 및 이를 제조하는 방법
CN112941098B (zh) 拟南芥花药绒毡层启动子表达载体及其构建方法与应用
CN112080523A (zh) 携带血管内皮生长因子及肝细胞生长因子双基因腺病毒载体的构建
CN108976293B (zh) 荔枝花色素苷生物合成负调控基因及其应用
CN109266686A (zh) 一种基因组核苷酸定点替换的方法
CN113481216B (zh) 多主棒孢菌CcTLS1蛋白与编码基因及其应用
CN109265562B (zh) 一种切刻酶及其在基因组碱基替换中的应用
CN113881670B (zh) 抗大豆花叶病毒的转基因植物构建方法
KR101760620B1 (ko) Hd1 인트론을 포함하는 식물 형질전환용 재조합 벡터 및 이의 이용
CN113215160A (zh) 一种植物来源启动子、表达载体及应用
CN112852828A (zh) 一种植物花粉管液泡荧光标记构建方法与应用

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant