CN105754964A - 水稻组蛋白甲基转移酶及其编码基因与应用 - Google Patents

水稻组蛋白甲基转移酶及其编码基因与应用 Download PDF

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CN105754964A
CN105754964A CN201610173430.2A CN201610173430A CN105754964A CN 105754964 A CN105754964 A CN 105754964A CN 201610173430 A CN201610173430 A CN 201610173430A CN 105754964 A CN105754964 A CN 105754964A
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沈文辉
刘坤朋
董爱武
俞瑜
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Abstract

本发明属于生物基因工程技术领域，具体为一种水稻组蛋白甲基转移酶及其编码基因与应用。该甲基转移酶是下述氨基酸残基序列之一：（1）序列表中的SEQ ID No：1；（2）将序列表中的SEQ ID No:1的氨基酸残基序列经过一至五十个氨基酸残基的取代和/或缺失和/或添加且对植物的生长发育具有调控作用的蛋白质。本发明还包括含有本发明基因的重组表达载体、转基因细胞系和工程菌以及扩增该基因中任一片段的引物对。该水稻组蛋白赖氨酸甲基转移酶的编码基因的反义转基因株系生长矮小，开花时间推迟。本发明为植物品种改良提供了一个优质基因，具有较高的实际应用价值，应用前景广阔。

Description

水稻组蛋白甲基转移酶及其编码基因与应用
技术领域
本发明属于生物基因工程技术领域，具体涉及一种与水稻表观遗传学调控相关的组蛋白甲基转移酶及其编码基因与应用。
背景技术
与可以活动的动物不同，营固着生长的植物其生长发育和环境应答等过程都与基因的转录表达调控密切相关。基因的转录表达调控除了受转录因子等遗传因素影响外还受到遗传辅因子等表观遗传因子的调控。近几年，表观遗传学（epigenetics）已然成为生命科学研究的热点领域。在基因组中除了DNA序列，还有许多调控基因的信息，它们本身不改变基因的序列，但是可以通过基因修饰，从而影响和调节基因的功能和特性，并且通过细胞分裂和增殖遗传下去。组蛋白，作为染色质基本单位--核小体的核心组成部分，其特定位点的赖氨酸甲基化修饰是重要的表观遗传学修饰之一。一般认为，组蛋白H3N末端第9、27位赖氨酸（H3K9和H3K27）、组蛋白H4N末端第20位赖氨酸（H4K20）与转录抑制相关，组蛋白H3N末端第4和36位赖氨酸（H3K4和H3K36）与转录激活相关。
与基因转录激活相关的组蛋白H3N末端4位赖氨酸（H3K4）的甲基化，通常被认为具有重要的生物学功能。然而其对植物生长发育的调控作用的研究近两年才刚刚开始。与真菌和动物中仅存在一个或几个组蛋白H3K4甲基转移酶(histonelysinemethyltransferase，HMTase)不同，植物中一般存在多个H3K4甲基转移酶。根据chromDB(http://www.chromdb.org/)的注释，在拟南芥基因组中有12个基因编码的蛋白质与真菌和动物中的H3K4甲基转移酶同源。已有研究表明，组蛋白H3K4甲基化修饰在植物生长发育过程中起着至关重要的作用。其中，SDG2是主效的H3K4甲基转移酶，sdg2突变体表现出植株矮小、纯合不育、雌雄配子体发育异常等多效性的表型（Berr,A,McCallum,E.J,Menard,R,Meyer,D,Fuchs,J,Dong,A.andShen,W-H.ArabidopsisSETDOMAINGROUP2isrequiredforH3K4trimethylationandiscrucialforbothsporophyteandgametophytedevelopment.PlantCell,22,3232-3248.）。sdg2突变体除了表现出育性相关表型，还具有多种表型，包括昼夜节律紊乱，激素应答缺陷等。进一步研究表明，SDG2还通过参与拟南芥主根干细胞的维持和侧根干细胞的形成，从而调控拟南芥根的生长发育（Yao,X.,Feng,H.,Yu,Y.,Dong,A.,andShen,W.-H..SDG2-MediatedH3K4MethylationIsRequiredforProperArabidopsisRootGrowthandDevelopment.PLoSOne,8,e56537.）。
拟南芥SDG2对生长发育的影响让我们思索水稻中它们的同源物是否具有同样重要的功能。与双子叶植物拟南芥不同，水稻是单子叶植物的一个典型代表。双子叶和单子叶植物,虽然可能基因间具有比较高的保守性,但它们的生长发育的调控机制可能存在很大差异。由于水稻生命周期长等原因,相关研究相对于拟南芥来说还相当滞后，水稻中组蛋白H3K4甲基化修饰的研究目前还很少。而研究水稻的价值，不仅在于它巨大的经济价值，还在于它是研究同为单子叶禾本科植物玉米、小麦、高粱等其他作物的理想模型。可以为我国的水稻分子育种提供理论基础和种质资源。
发明内容
本发明的目的是提供一个来源于水稻的组蛋白甲基转移酶及其编码基因与应用。
本发明所提供的组蛋白甲基转移酶，名称为SDG701，来源于水稻（Oryzasativassp.japonica），是下述氨基酸残基序列之一：
（1）序列表中的SEQIDNo:1；
（2）将序列表中的SEQIDNo:1的氨基酸残基序列经过一至五十个氨基酸残基的取代和/或缺失和/或添加且对植物的生长发育具有调控作用的蛋白质。
序列表中的SEQIDNo:1由2216个氨基酸残基组成。
编码本发明组蛋白甲基转移酶的基因（SDG701），是下述核苷酸序列之一：
（1）序列表中的SEQIDNo:2的核苷酸序列；
（2）编码序列表中的SEQIDNo:1蛋白质序列的DNA；
（3）与序列表中的SEQIDNo:2限定的核苷酸序列具有90％以上同源性且编码相同功能蛋白质的核苷酸序列；
（4）在高严谨条件下可与序列表中的SEQIDNo:2限定的DNA序列杂交的核苷酸序列。
上述高严谨条件为：用0.1×SSPE（或0.1×SSC），0.1%SDS的溶液，在65℃下杂交并洗膜。
序列表中的SEQIDNo:2由6651个碱基组成，其编码框为自5’端第1-6651位碱基，编码具有序列表中的SEQIDNo:1的氨基酸残基序列的蛋白质。
本发明还包括含有本发明基因的重组表达载体、转基因细胞系和工程菌以及扩增该基因中任一片段的引物对。
本发明还提供上述的水稻组蛋白甲基转移酶基因在调控植物生长发育中的应用。
在实际应用中，将上述水稻组蛋白甲基转移酶基因的反义表达载体转化水稻，得到生长矮小的水稻，使植物的生长发育得到调控。
上述重组表达载体均可按照常规方法构建。
本发明的水稻组蛋白甲基转移酶基因的反义转基因植株株系生长矮小，开花时间推迟。本发明为植物品种的改良提供了一个优质基因，同时也对深入理解组蛋白甲基转移酶的生物学功能和生物学过程，进一步破译组蛋白密码和理解组蛋白密码在高等植物生长发育、遗传变异中的作用具有非常重要的意义。本发明的蛋白质及其编码基因具有较高的实际应用价值，应用前景广阔。
附图说明
图1为水稻组蛋白甲基转移酶SDG701的蛋白活性测定结果。
图2为野生型水稻（WT）和T0代转反义SDG701基因水稻（701Ri-1-1，701Ri-1-2，701Ri-2-1，701Ri-2-2）的表型图。
图3为野生型水稻（WT）和转反义SDG701基因水稻（701Ri-1-1，701Ri-1-2，701Ri-2-1，701Ri-2-2）中SDG701基因表达的实时荧光定量PCR检测结果。
具体实施方式
下述实施例中的方法，如无特别说明，均为常规方法。所用引物和测序工作由上海桑尼生物科技有限公司完成。
实施例1.水稻组蛋白甲基转移酶基因SDG701的获得
对GenBank中公布的拟南芥的SDG2的基因序列（GenBank号：NC_003075.7）通过同源序列比较，从水稻基因组中获得同源序列，并根据该同源序列的5’和3’末端序列设计一对引物，引物序列分别为：5'-ATGCCGGACAAGGGGGAGA-3'（SEQIDNo:3），和5'-CTAGCCTAGGAAGACGTTGGATCTTG-3'（SEQIDNo:4）。
提取水稻黄化苗总RNA（Promega，SVtotalRNAisolationsystem），以水稻的总RNA为模板，用AMV反转录酶（TaKaRa）合成cDNA（依据PlantRT－PCRKit2.01（TaKaRa）的用户手册进行）。以cDNA为模板，PCR扩增水稻组蛋白甲基转移酶基因SDG701的全长cDNA序列。50μlPCR反应体系包含：模板2μl，高保真酶KODplus（TOYOBO）1μl，10×缓冲液5μl，2.5μMdNTP8μl，20μM的5'和3’引物各1μl，水32μl。反应条件为：94℃预变性2分钟；94℃变性30秒，55℃退火30秒，68℃延伸7分钟，共30个循环。反应结束后，将PCR产物进行0.8％琼脂糖凝胶电泳检测，回收并纯化约6651bp的扩增片段，将其克隆到载体pUC19（TaKaRa）中，得到含有回收片段的重组质粒，经测序表明水稻组蛋白甲基转移酶基因SDG701全长cDNA具有序列表中的SEQIDNo:2的核苷酸序列，序列表中的SEQIDNo:2由6651个碱基组成，其编码框为自5’端第1-6648位碱基，编码具有序列表中的SEQIDNo:1的氨基酸残基序列的蛋白质，编码蛋白命名为SDG701。
实施例2.水稻组蛋白甲基转移酶基因SDG701的蛋白活性测定
以实施例1得到的pUC19-SDG701为模板，PCR扩增SDG701自5’端第4035－6648位的核苷酸序列（包含组蛋白甲基转移酶的催化活性中心――SETdomain结构域），5'和3’引物分别为：5'-cccgggCCAAGGTTGATTTCTGATAG-3'（SEQIDNo:5），和5'-gcggccgcCTAGCCTAGGAAGAC-3'（SEQIDNo:6）。在50μlPCR扩增体系中包含模版pUC19-SDG701200ng，5'和3’引物各20pmol，10×buffer5μl，dNTP各20μM，高保真KODplus（TOYOBO）1μl。按以下条件进行扩增：94℃预变性2分钟；94℃变性30秒、55℃退火30秒、68℃延伸3分钟，共30个循环。PCR产物经SmaⅠ和NotI酶切位点克隆到大肠杆菌表达载体pGEX-4T1（GElifesciences），测序验证其正确后得到大肠杆菌表达质粒pGEX-4T1-SDG701C。
将GST负对照和GST融合的SDG714正对照以及GST融合的SDG701C重组蛋白在大肠杆菌中的表达纯化依据GlutathioneSepharose4FastFlow（GElifesciences）的用户手册进行。将连有GST，GST-SDG714或者GST-SDG701C蛋白的Glutathione-Sepharosebeads经洗涤后直接用于酶活测定实验。
组蛋白甲基转移酶的体外活性测定按Rea等（Nature，2000，406：593–599）的方法进行：在30μlMAB测活缓冲液（50mMTris-HCl（pH8.5），20mMKCl，10mMMgCl2，250mMSucrose，100μMZnCl2，10mMβ-mercaptoethanol）中，加入的酶（GST－SDG725set），底物（核心组蛋白H2A、H2B、H3和H4，购自Millipore）和250nCimethyl-14C-SAM（GElifesciences），混合均匀，37℃反应1小时。反应产物经15%SDS－PAGE分离并染色脱色，凝胶于70℃真空干燥2hrs后放射自显影。
图1中上图为SDS－PAGE电泳结果，下图为放射自显影结果，GST蛋白作为阴性对照，正对照为一已知的组蛋白H3的甲基转移酶GST-SDG714。结果显示GST－SDG701C具有组蛋白H3K4特异的甲基转移酶活性。
实施例3.检测SDG701对水稻生长发育的调控作用
一、SDG701反义表达载体的构建
利用RNA干扰(RNAinterference,RNAi)技术，以SDG701基因为靶基因，构建RNA干扰载体。以实施例1得到的pUC19-SDG701为模板，分别以两对不同的引物PCR扩增SDG701自5’端第98到463位的核苷酸序列，作为发夹结构的互补DNA双链。所用的引物对1为：5'-ccatggggatccGGGAGCTGCTGCTCAATGGGGAG-3'（SEQIDNo:7）和5'-gaattcCCGACGAGCCGAGCTTCTT-3'（SEQIDNo:8），所得PCR产物为fSDG701i-1（forward）。引物对2为5'-tctagaGGGAGCTGCTCAATGGGGAG-3'（SEQIDNo:9）和5'-aagcttCCGACGAGCCGAGCTTCTT-3'（SEQIDNo:10），所得PCR产物为rSDG701i-1（reversed）。为了便于后续的载体构建，在fSDG701i-1的5’和3’端分别添加了NcoⅠBamHⅠ和EcoRⅠ的限制性酶切位点，在rSDG701i-1的5’和3’端分别引入了XbaI和HindIII的限制性酶切位点。
为了确保干扰的效果，我们同时以实施例1得到的pUC19-SDG701为模板，分别以两对不同的引物PCR扩增SDG701自5’端第519到835位的核苷酸序列，作为发夹结构的互补DNA双链。所用的引物对1为：5'-ccatggggatccCAGGGCAACAGAGCCTGAGA-3'（SEQIDNo:11）和5'-gaattcGGGATGGGCGCATTCTGTGG-3'（SEQIDNo:12），所得PCR产物为fSDG701i-2（forward）。引物对2为5'-tctagaCAGGGCAACAGAGCCTGAGA-3'（SEQIDNo:13）和5'-aagcttGGGATGGGCGCATTCTGTGG-3'（SEQIDNo:14），所得PCR产物为rSDG701i-2（reversed）。为了便于后续的载体构建，在fSDG701i-2的5’和3’端分别添加了NcoⅠBamHⅠ和EcoRⅠ的限制性酶切位点，在rSDG701i-2的5’和3’端分别引入了XbaI和HindIII的限制性酶切位点。
为了便于RNAi载体的构建，在连接正反两条DNA分子的PDK内含子（由法国植物分子生物学研究所赠送）两端通过PCR方法分别引入了HindIII和EcoRⅠ的限制性酶切位点，构建了中间载体pGEMT，所用引物为：5’－aaaagcttccaatttggtaaggaaataatt－3’（SEQIDNo:15）和5’－aagaattctttcgaacccagcttccc－3’（SEQIDNo:16）。
将fSDG701i-1和fSDG701i-2经NcoⅠ和EcoRⅠ双酶切，连接入NcoⅠ和EcoRⅠ双酶切过的含PDK内含子的中间载体pGEMT中。接着用XbaI和HindIII双酶切rSDG701i-1和rSDG701i-2，分别连接入经XbaI和HindIII双酶切的pGEMT-fSDG701i-1-PDKintron和pGEMT-fSDG701i-2-PDKintron载体中。最后用BamHⅠ和XbaI双酶切pGEMT-fSDG701i-1-PDKintron-rSDG701i-1和pGEMT-fSDG701i-2-PDKintron-rSDG701i-2连接入经BamHⅠ和XbaI双酶切的pHB载体（由中科院上海植物生理研究所林鸿宣课题组赠送）中，即得RNA干扰载体pHB－fSDG701i-1－PDKintron－rSDG701i-1和pHB－fSDG701i-2－PDKintron－rSDG701i-2。
二、SDG701反义表达载体转化水稻植株
参照Hiei等（PlantMolBiol，1997，35：205－218）的方法转化水稻。将质粒pHB－fSDG701i-1－PDKintron－rSDG701i-1和pHB－fSDG701i-2－PDKintron－rSDG701i-2通过电激法分别转入根瘤农杆菌EHA105中，经筛选获得阳性克隆。将携带质粒pHB－fSDG701i-1－PDKintron－rSDG701i-1和pHB－fSDG701i-2－PDKintron－rSDG701i-2的农杆菌EH105分别接种到5ml含100mg/L卡那霉素的YEB液体培养基中28℃摇菌培养至对数生长期晚期，再1：100扩大培养到OD600为0.5左右。收集农杆菌菌体并重悬于转染培养基中。按照常规方法浸染水稻日本晴的愈伤组织，经共培养感染、抗性培养基筛选及分化培养基上分化得到潮霉素抗性的阳性苗。待小苗长至10cm左右时，将小苗移至室外网室种植，即为T0代植株，结果如图2所示。
并且能结籽的T0代植株的后代在T1代和T2代都表现出明显的生长矮小，开花时间推迟的表型，进一步确定了该转基因性状稳定遗传，。
三、荧光定量PCR检测转反义SDG701基因水稻中SDG701的表达
在相同条件下对转反义SDG701基因水稻和野生型水稻进行培育。按实施例1中相同方法提取水稻总RNA并且反转录合成cDNA。然后用荧光定量PCR检测转反义SDG701基因水稻和野生型水稻中SDG701的表达情况。检测SDG701所用引物P1为：5'-TGCTGGTTTGCCTGGATGGC-3'（SEQIDNo:17），5'-ACTGGCGCTTCTCGTCCACA-3'（SEQIDNo:18）。检测SDG701所用引物P2为：5'-CACTGCTCACCTGGTGCGGTT-3'（SEQIDNo:19），5'-CGGCCTGAACCTCGGCATCA-3'（SEQIDNo:20）。结果如图3所示，相对于野生型水稻，SDG701的表达在转反义SDG701基因水稻中呈现明显的下降，且SDG701表达下调的程度与表型强度呈明显的正相关，进一步证明转反义SDG701基因水稻的表型缺陷是由于SDG701的表达下降而引起的。
SEQUENCELISTING
<110>复旦大学
<120>水稻组蛋白甲基转移酶及其编码基因与应用
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aaggggacaacgcttcctgtcgttaagataaatgcccgtgttgttaaggaccaatcctct2400
tctgaactccgtataaagcccaaagtagctgatcggccaccttctagatcttcgcgctcc2460
cattcaattgggactgataggagttctacccatgaaggttcgtctcattctaagaaacat2520
catgaacacgattcacaaagtttgcacaagtccaagtctgttccaaacattccaaaggac2580
catgtatgtactgttgaagaattgtcagtcaaagtaggtgactggtactacctggatgga2640
actggtcatgagcgtggcccattttcttattctgaattgcaagaattagctaagaagggc2700
actatccttgaagggagcagtgttttccggaagattgataacacatggcttccagtgctt2760
aaggatttaaaatctggttgctctgctcggaatggggaggctggaagttctacctctgct2820
ctcacacactcaaaccagtccaacttccacgagatgcatccacagtttgtgggttatact2880
cgtggtaaactgcatgaactagtcatgaagtatttcaagagcagggaacttactttggct2940
ataaatgagaattttgtgcaagaggatggcgggtccacaaaaagggcaaggttgctgcct3000
gatcaaagtgatgaatatactgatatgtctgaggacattcttgccagtcagaaggatgat3060
tgttgttttgaagacttatttgagggggctgctcatgttaaagaaagccctctgaattcc3120
agaacggaaagtgaaagctggggtttattaaatgaacatgtgttagcaagaattttccat3180
tttctgagggcagacgtgaagtcacttatttcttctgcagctacctgtagctggtggaac3240
actgcagccaagtattacaggagtgtttgtagattcattgatttgtcttctttgggccct3300
cagtgcactgataatgttttccacgacatcatggctggttatgacatgcaaaatattaga3360
acacttgttttaacagggtgttcaaatcttagttctcttgcccttgcggaagtacttaag3420
cgttttccgcacatatcatatgtgcacattcaaggttgcagtcagctaggggatcttaaa3480
aacaaatttcagcatgtaaaatggattaagagctctttgaatccagatgcatcataccag3540
aaaattagaagcttgaagcagatagatgatgggagcaacagcacatccaaagctggaagg3600
atcttgaccagtcagatgggtggttctgatgagcttgatggctattttgctgatatttca3660
aatagagagagttccaccctttcctttggacaagggttctataaacgatcaaagtggctt3720
gatattagaaagtcgtctgcggttttgtcaagggatgcacagatgaggcgtttgatgcag3780
cggaaggccgagaatagctaccggaagatggaagagtttgtcatcaacaaactaaaagaa3840
atcatgaagagcagtagatttgatttttttgttccaaaggttgcgaaaattgaagttagg3900
ttgaaaaatggatactatgctcgccatggctttagttatatcaagaatgacatccgttct3960
atgtgccgagatgcattaagatataaaggtcgaagtgacttgggagatatgaaacagatt4020
gtcgtggccttcatccagctagcaaagaaacttgagaacccaaggttgatttctgataga4080
gatgggacagcagttcagaaggacagttctgacatgagtcagtattcttcagatctgaaa4140
ctcaaaaagaaacagagtaaaactatgtcagaaagaaggggagcaaactggactactgct4200
ggagcagatccgtcatctcgtgcatttgaccgtgaaatcaaaagaagtctttctaaatta4260
aagaaaagggatatcgattctggtagtgaaacatctgatgatgatgatggctactctgaa4320
ggtgatgagactgagagtgaaactactgtttctgatacagagagtgatcttgatgtaaat4380
tcgggagcatgggatttaaagggaaatggtatgaagttatttgaatcaagcgagtctttg4440
acggatgatcgtggatggggtgctcgcatgacaaaagcaagccttgttcccccagtgact4500
aggaagtatgaggttattgagaagtacctaattgtagcagatgaggaggaagtactgaga4560
aagatgcgggtcgctttacctgatgactattcagagaaactgctctcgcagaagaacggc4620
actgaaaatttggaactaccagaggttaaggattatcaacctagaaaagtacctggggat4680
gaagttcttgagcaagaagtgtatggaatagatccatatacacataatctcctacttgaa4740
atgatgcctactgaacttgattggccatcctctgacaaacatacctttgttgaagagttg4800
cttctgaacaccttgaataagcaagtgaggcagttcactggttctggaaatactcccatg4860
gtttacccccttaaacctgtaattgaagaaatccaaaagtctgcagaggagagtggtgac4920
agacggacttcaaagatgtgccttggaatgctaaaggccatgaggaaccacccagaatat4980
aactatgtcgcctataggaagggtcttggagttgtttgtaacaaaacgggtggatttggt5040
gtagacgactttgtcatcgagttctttggggaggtatacccttcttggagatggtatgaa5100
aaacaagatggcattaagcatatacaaaacaatagcgacgatcaagctcctgagttttac5160
aacattatgctagaacggccaaagggagaccgtgatgggtatgacttggttttcgttgat5220
gcaatgcataaggctaactatgcgagcagaatttgtcactcctgcaaccctaactgtgaa5280
gcaaaagtaacggctgtggatggtcattatcagattggaatttacactgttcgaccaatt5340
gcagaaggcgaggaaatcacttttgattacaactctgttactgagagtaaagaagagcac5400
gaagcgtcagtttgtctctgtggaagtcaaatatgcaggggcagctatttaaatttttct5460
ggggaaggagcttttgaaaaggtcttgatggaattccatggtgtgctcgatcgacatagt5520
cttctgttacaggcttgtgaagcaaactctgtttctcaacaagacttaattgacttgggg5580
agagctggtcttggtacctgtttgcttgctggtttgcctggatggcttgttgcttacact5640
gctcacctggtgcggtttatattctttgagagacagaaacttcctcatgagatctttaaa5700
cacaatgtggacgagaagcgccagtttttcacagatataaacatggattccgagaagaat5760
gatgccgaggttcaggccgagggagttctgaattcaagattacagaatttgactcataca5820
cttgacaaggtaagatatgtcatgagatgcatatttggggatcccaagaatgctcctcct5880
ccactggtgaggctaactgggagaagtctagtttctgccatctggaaaggggaaggctcg5940
ttagttgatgaacttcttgagtcaatggaacctcatgttgaggaagatgtacttactgac6000
ctcaaagccaaaattcgtgctcatgatccatctggttccgaggacattgagggagaaatc6060
cgaagttctcttttatggttgcgtgatgagctgagaactctttcatgcacatacaagtgt6120
cggcatgatgctgctgctgatttgattcacatgtatgcttacacaaagtgttttttcaga6180
gtccgagattacaagacggtaaaatctccaccggttcttatcagtcctcttgatttaggt6240
cccaaatatgcagataaactgggaccaggcttccaggagtactgtaagacatacccagaa6300
aattattgcttaggtcaactcatttattggtatagccagaatgcggagcctgaatctaga6360
ttgacaagagctagaaagggttgtatgtcattgccagatgtatcctccttctatgtgaag6420
tcagtaaaaccaacacaagagcgagtctatggcagcagaactgtgagattcatgttagcg6480
cgcatggagaaccaggcacaaaggccatggcccaaggaccggatatgggttttcaagagt6540
gacccaagattctttggtactccaatgatggatgccgtattgaataattccccccttgac6600
aaggagatggtgcattggctcaagacaagatccaacgtcttcctaggctag6651
<210>3
<211>19
<212>DNA
<213>
<400>3
atgccggacaagggggaga19
<210>4
<211>26
<212>DNA
<213>
<400>4
ctagcctaggaagacgttggatcttg26
<210>5
<211>26
<212>DNA
<213>
<400>5
cccgggccaaggttgatttctgatag26
<210>6
<211>23
<212>DNA
<213>
<400>6
gcggccgcctagcctaggaagac23
<210>7
<211>35
<212>DNA
<213>
<400>7
ccatggggatccgggagctgctgctcaatggggag35
<210>8
<211>25
<212>DNA
<213>
<400>8
gaattcccgacgagccgagcttctt25
<210>9
<211>26
<212>DNA
<213>
<400>9
tctagagggagctgctcaatggggag26
<210>10
<211>25
<212>DNA
<213>
<400>10
aagcttccgacgagccgagcttctt25
<210>11
<211>32
<212>DNA
<213>
<400>11
ccatggggatcccagggcaacagagcctgaga32
<210>12
<211>26
<212>DNA
<213>
<400>12
gaattcgggatgggcgcattctgtgg26
<210>13
<211>26
<212>DNA
<213>
<400>13
tctagacagggcaacagagcctgaga26
<210>14
<211>26
<212>DNA
<213>
<400>14
aagcttgggatgggcgcattctgtgg26
<210>15
<211>30
<212>DNA
<213>
<400>15
aaaagcttccaatttggtaaggaaataatt30
<210>16
<211>26
<212>DNA
<213>
<400>16
aagaattctttcgaacccagcttccc26
<210>17
<211>20
<212>DNA
<213>
<400>17
tgctggtttgcctggatggc20
<210>18
<211>20
<212>DNA
<213>
<400>18
actggcgcttctcgtccaca20
<210>19
<211>21
<212>DNA
<213>
<400>19
cactgctcacctggtgcggtt21
<210>20
<211>20
<212>DNA
<213>
<400>20
cggcctgaacctcggcatca20

Claims (10)

1.水稻组蛋白甲基转移酶，其特征在于来源于水稻，名称为SDG701，是具有下述氨基酸残基序列之一的蛋白质：
（1）SEQIDNo:1；
（2）将SEQIDNo:1的氨基酸残基序列经过一至五十个氨基酸残基的取代和/或缺失和/或添加且对植物的生长发育具有调控作用的蛋白质。
2.编码权利要求1所述的组蛋白甲基转移酶的基因。
3.根据权利要求2所述的基因，其特征在于：所述基因是下述核苷酸序列之一：
（1）SEQIDNo:2的核苷酸序列；
（2）编码SEQIDNo:1蛋白质序列的DNA；
（3）与SEQIDNo:2限定的核苷酸序列具有90％以上同源性且编码相同功能蛋白质的核苷酸序列；
（4）在高严谨条件下可与SEQIDNo:2限定的DNA序列杂交的核苷酸序列。
4.含有权利要求2或3所述的水稻组蛋白甲基转移酶基因的表达载体。
5.含有权利要求2或3所述的水稻组蛋白甲基转移酶基因的转基因细胞系。
6.含有权利要求2或3所述的水稻组蛋白甲基转移酶基因的工程菌。
7.扩增权利要求2或3所述的水稻组蛋白甲基转移酶基因中任一片段的引物对。
8.权利要求2或3所述的水稻组蛋白甲基转移酶基因在植物生长发育中的应用。
9.根据权利要求7所述的应用，其特征在于，所述的植物包括单子叶植物和双子叶植物。
10.根据权利要求7所述的应用，其特征在于，将所述水稻组蛋白甲基转移酶基因的反义表达载体转化水稻，得到生长矮小，开花时间推迟的水稻。
CN201610173430.2A 2016-03-24 2016-03-24 水稻组蛋白甲基转移酶及其编码基因与应用 Pending CN105754964A (zh)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109053544A (zh) * 2018-08-24 2018-12-21 江苏理工学院 一种化合物b11作为组蛋白甲基转移酶nsd3活性抑制剂及其应用

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CN101792748A (zh) * 2010-03-25 2010-08-04 复旦大学 水稻的组蛋白赖氨酸甲基转移酶及其编码基因与应用
CN104402982A (zh) * 2014-12-03 2015-03-11 复旦大学 水稻组蛋白赖氨酸修饰识别蛋白及其编码基因与应用

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109053544A (zh) * 2018-08-24 2018-12-21 江苏理工学院 一种化合物b11作为组蛋白甲基转移酶nsd3活性抑制剂及其应用

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Application publication date: 20160713