CN108277228A - 一种提高植物对双生病毒抗性的方法 - Google Patents

一种提高植物对双生病毒抗性的方法 Download PDF

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CN108277228A
CN108277228A CN201611222101.9A CN201611222101A CN108277228A CN 108277228 A CN108277228 A CN 108277228A CN 201611222101 A CN201611222101 A CN 201611222101A CN 108277228 A CN108277228 A CN 108277228A
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叶健
孙艳伟
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Abstract

本发明提供一种改造的抗双生病毒的vTALE分子，和利用编码该分子的重组载体培育具有稳定的广谱双牛病毒抗性的植物的方法。其策略为利用高效改变双生病毒基因组特定区域的表观遗传学修饰从而干扰双生病毒的正常复制及转录过程，达到提高植物对双生病毒的免疫能力的目的。所述vTALE分子以植物病原细菌水稻黄单胞杆菌三型分泌系统效应因子AvrXa27基因序列为骨架序列，保留其N端和C端序列，然后对其DNA识别/结合结构域序列和功能输出结构域序列进行改造，其中所述改造后的DNA识别/结合结构域序列靶向SEQ ID NO：1所示的IR‑vTALE靶点序列或SEQ ID NO：2所示的RB‑vTALE靶点序列，并且所述改造后的功能输出结构域序列选自由编码SEQ ID NOs：8‑12所示的氨基酸序列的核苷酸序列组成的组中的任一种。

Description

一种提高植物对双生病毒抗性的方法
技术领域
本发明涉及分子生物学领域，特别涉及一种改造的抗双生病毒的vTALE分子，尤其涉及一种对双生病毒科病毒具有广谱抗性且有效稳定的vTALE分子。本发明还涉及所述vTALE分子的应用以及利用所述vTALE分子培育具有稳定的广谱双生病毒抗性的植物的方法。
背景技术
双生病毒(Geminivirus)为孪生颗粒形态的单链DNA病毒，病毒颗粒大小约为20×30nm。依据病毒的基因组结构、寄主范围、传播介体以及基因组序列的相关性将双生病毒分为七个属，即菜豆金色花叶病毒属(Begomovirus)、玉米线条病毒属(Mastrevirus)、甜菜曲顶病毒属(Curtovirus)、番茄伪曲顶病毒属(Topocuvirus)、伊朗甜菜曲顶病毒属(Becurtovirus)、芜菁曲顶病毒属(Turncurtovirus)和弯叶画眉草条纹病毒属(Eragrovirus)。双生病毒高度依赖于寄主植物细胞的DNA复制和RNA转录系统来完成自身的复制和转录过程。
在最近20年间，双生病毒引起的病害由局部发生的小病害衍变成目前全球性的最重要的植物病毒病害之一，危害玉米、小麦、棉花、木薯、番茄等重要农作物和经济作物，仅对于木薯，每年在全球造成的经济损失就达数十亿美元之巨。在我国，随着入侵烟粉虱的不断蔓延，导致由其传播的双生病毒病害——番茄黄化曲叶病在18个省市多个番茄产区发生，成为我国番茄生产上最重要的病害之一。2009年经济损失超过50亿元，2010--2012年危害进一步扩大，病害发生面积超过100万亩。因此，对双生病毒病害的有效防治已经成为亟待解决的实际生产问题。
目前，在实际生产中仍普遍采用化学防治的方法来直接或者间接地防控双生病毒病害，但效果并不理想，同时化学防治对环境及食品安全也构成了严重的威胁。随着分子生物学的不断发展，科学家们对植物抗双生病毒的分子机制有了进一步的解析，研究较清楚的机制主要有两类：抗性较专一的抗性基因(R gene)介导的抗性和抗性谱较宽的RNA沉默(RNA silencing)介导的抗性。利用抗性基因选育抗性品种来防控双生病毒的方法具有育种周期长、抗病谱较窄的弱点，而双生病毒的突变率高、病毒同入侵型介体昆虫形成互惠共生的关系等原因导致该种方法未能快速、有效地发挥防控作用。而利用RNA沉默机制来防控双生病毒的方法由于存在抗性不稳定且易丢失，此外双生病毒同其他病毒一样也进化出了有效的抑制植物RNA沉默的基因沉默抑制子(Suppressor of RNA silencing)，使得该方法也未能在实际生产中得到广泛应用。
最近发展出来一种利用CRISPR/Cas技术使植物获得抗双生病毒的新方法。此系统的工作原理是crRNA(CRISPR-derived RNA)通过碱基配对与tracrRNA(trans-activatingRNA)结合形成tracrRNA/crRNA复合物，此复合物引导核酸酶Cas9蛋白在与crRNA配对的序列靶位点剪切双链DNA。而通过人工设计这两种RNA，可以改造形成具有引导作用的sgRNA(short guide RNA)，足以引导Cas9对DNA的定点切割。但是研究表明通过该方法植物获得的对双生病毒的抗性并不稳定且常有脱靶的现象。
综上所述，利用分子生物学技术发展一种有效并且抗性稳定的抗双生病毒的新方法对于农作物及经济作物的高产和稳产以及环境和食品的安全都是十分必要的。
发明内容
本发明的目的是提供一种对双生病毒科病毒具有广谱抗性且有效稳定的vTALE分子。
本发明提供的对双生病毒科病毒具有广谱抗性且有效稳定的vTALE分子是一种改造的抗双生病毒的vTALE分子，其以植物病原细菌水稻黄单胞杆菌(Xanthomonas oryzaepv.oryzae，缩写为Xoo)三型分泌系统效应因子AvrXa27基因序列为骨架序列，保留AvrXa27基因的N端和C端序列，然后对AvrXa27基因的DNA识别/结合结构域序列和功能输出结构域序列进行改造而获得。其中改造的部分涉及DNA识别/结合结构域序列和功能输出结构域序列两部分(下文将这两部分分别称为“改造后的DNA识别/结合结构域序列”和“改造后的功能输出结构域序列”)。
vTALE分子的DNA识别/结合结构域经人工设计能够特异性地识别并结合任意指定的DNA序列，在功能输出结构域的作用下从而可以实现对靶基因的调控与修饰。通过对双生病毒基因组特定区域进行人工设计的表观遗传学修饰从而干扰双生病毒的正常复制及转录过程，达到提高植物对双生病毒的免疫能力的目的。
本发明分别针对双生病毒科多株双生病毒高度保守的IR(Intergenetic Region)区域设计了vTALE靶点IR-vTALE及针对菜豆金色花叶病毒属病毒甘蓝曲叶病毒(Cabbageleaf curl virus，CaLCuV)复制相关蛋白结合位点(Replication associated proteinBinding，Rep Binding)设计了特异性的RB-vTALE靶点为vTALE的DNA识别/结合结构域的靶点序列，功能输出结构域融合表达表观遗传学修饰相关因子。
在第一方面，本发明提供一种改造的抗双生病毒的vTALE分子，其以植物病原细菌水稻黄单胞杆菌(Xanthomonas oryzae pv.oryzae)三型分泌系统效应因子AvrXa27基因序列为骨架序列，保留AvrXa27基因的N端和C端序列，然后对AvrXa27基因的DNA识别/结合结构域序列和功能输出结构域序列进行改造而获得，其中所述改造后的DNA识别/结合结构域序列靶向SEQ ID NO：1所示的IR-vTALE靶点序列或SEQ ID NO：2所示的RB-vTALE靶点序列，并且所述改造后的功能输出结构域序列选自由编码SEQ ID NOs：8-12所示的氨基酸序列的核苷酸序列组成的组中的任一种。这两部分序列可以通过特异的限制性内切酶处理后连接。本领域技术人员能够选择这两个序列中合适的特异性限制性内切酶酶切位点经限制性内切酶处理后将二者连接在一起。
优选地，所述改造后的DNA识别/结合结构域序列靶向IR-vTALE靶点序列，其序列为5’-TAATATTACCGGATGGCC-3’(SEQ ID NO：1)。
优选地，所述改造后的DNA识别/结合结构域序列靶向RB-vTALE靶点序列，其序列为5’-TATATATTGGACACCAAG-3’(SEQ ID NO：2)。
优选地，所述改造后的功能输出结构域序列为编码AtKYP(其氨基酸序列为SEQ IDNO：8)的核苷酸序列。
优选地，所述改造后的功能输出结构域序列为编码AtHDAC19(其氨基酸序列为SEQID NO：9)的核苷酸序列。
优选地，所述改造后的功能输出结构域序列为编码AtMET1(其氨基酸序列为SEQID NO：10)的核苷酸序列。
优选地，所述改造后的功能输出结构域序列为编码AtSRDX(其氨基酸序列为SEQID NO：11)的核苷酸序列。
优选地，所述改造后的功能输出结构域序列为编码AtSUVH2(其氨基酸序列为SEQID NO：12)的核苷酸序列。
优选地，所述改造后的功能输出结构域序列是选自由SEQ ID NOs：3-7所示的核苷酸序列组成的组的任意一种核苷酸序列。
根据本发明第一方面构建的抗双生病毒的vTALE分子包括，但不限于，SEQ IDNOs：17-19和26-30。其中，优选的vTALE分子的序列为IR-vTALE-AtKYP(SEQ ID NO：17)和RB-vTALE-AtKYP(SEQ ID NO：26)。其中IR-vTALE-AtKYP(SEQ ID NO：17)包括来源于Xoo的AvrXa27基因N端序列、靶向IR-vTALE靶点序列的序列、来源于Xoo的AvrXa27基因C端序列和AtKYP氨基酸序列(见图13)；RB-vTALE-AtKYP(SEQ ID NO：26)包括来源于Xoo的AvrXa27基因N端序列、靶向RB-vTALE靶点序列的序列、来源于Xoo的AvrXa27基因C端序列和AtKYP氨基酸序列(见图22)。
本领域技术人员知晓，现有技术中已有的一般TALE分子主要由两部分组成：DNA识别/结合结构域(Recognition/Binding domain)和激活结构域(Activation domain，AD)(图1A)。其中DNA识别/结合结构域是由一段串联排列的重复序列组成，能够特异性识别并结合靶基因序列，每个重复序列由长度为33-35个氨基酸组成，除了第12和13位的氨基酸可变外，其他氨基酸序列高度保守，而重复序列可变的双氨基酸残基(Repeat variable di-residue，RVD)是TALE实现靶向识别特异DNA碱基的关键位点，不同的RVD能够相对特异地识别A、T、C、G四种碱基中的一种或多种，其中分别与这4种碱基相对应的最常见的4种RVD分别是NI＝A，NG＝T，HD＝C和NN＝G或A。激活结构域能够在TALE特异性识别并结合靶基因后激活靶基因的表达。
本发明设计的vTALE分子结构示意图显示在图1B中。本发明设计的vTALE分子的DNA识别/结合结构域所结合的靶点序列选自下述中的任一种：针对双生病毒科多株病毒高度保守的IR(Intergenetic Region)区域序列设计的广谱抗双生病毒科病毒的Intergenetic Region-vTALE(IR-vTALE)靶点和针对菜豆金色花叶病毒属病毒甘蓝曲叶病毒(Cabbage leaf curl virus，CaLCuV)复制相关蛋白(Replication associatedprotein，Rep)结合位点设计的vTALE靶点Rep Binding-vTALE(RB-vTALE)。此外，由于TALE蛋白所识别的靶点DNA序列必须以T开始，所以，虽然两个靶点序列都只含有16.5个串联排列的重复序列，但是都可以特异识别并结合18bp的靶点DNA序列。根据本发明人设计的靶点DNA序列(即，IR-vTALE靶点序列或RB-vTALE靶点序列)，本领域技术人员能够设计相应的氨基酸重复序列进行串联，构成改造的DNA识别/结合结构域，这是在本领域技术人员的技能范围之内的。本发明人的创造性劳动体现在对AvrXa27基因的DNA识别/结合结构域序列所靶向的靶点序列的选择。
在第二方面，本发明提供本发明改造的抗双生病毒的vTALE分子在培育具有稳定的广谱双生病毒抗性的植物的方法中的应用。
在第三方面，本发明提供一种用于培育具有稳定的广谱双生病毒抗性的植物的方法，所述方法包括用第一方面的改造的抗双生病毒的vTALE分子转化待培育的植物，使vTALE在所述植物中稳定表达。也就是说，使用第一方面的改造的抗双生病毒的vTALE分子对植物进行转基因处理，获得具有稳定的广谱双生病毒抗性的植物。
在一个优选的实施方案中，本发明提供一种用于培育具有稳定的广谱双生病毒抗性的植物的方法，所述方法包括下述步骤：
(1)vTALE分子的构建：所述vTALE分子以植物病原细菌水稻黄单胞杆菌(Xanthomonas oryzae pv.oryzae)三型分泌系统效应因子AvrXa27基因序列为骨架序列，保留AvrXa27基因的N端和C端序列，然后对AvrXa27基因的DNA识别/结合结构域序列和功能输出结构域序列进行改造而获得，其中所述改造后的DNA识别/结合结构域序列靶向SEQ IDNO：1所示的IR-vTALE靶点序列或SEQ ID NO：2所示的RB-vTALE靶点序列；并且所述改造后的功能输出结构域序列选自由编码SEQ ID NOs：8-12所示的氨基酸序列的核苷酸序列组成的组中的任一种；
(2)植物转化：将步骤(1)构建的vTALE分子，转化到所述植物中进行稳定表达。
优选地，所述改造后的DNA识别/结合结构域序列靶向SEQ ID NO：1所示的IR-vTALE靶点序列或SEQ ID NO：2所示的RB-vTALE靶点序列。
具体地，在初步验证实验中，本发明人发现，在本氏烟草中瞬时表达步骤(1)构建的vTALE分子的优选组合IR-vTALE-AtKYP分子能够使本氏烟草获得对双生病毒科病毒的广谱抗性。将含有IR-vTALE-AtKYP分子的农杆菌注射本氏烟草24h后，再接种双生病毒科不同属病毒的侵染性克隆，包括玉米线条病毒属(Mastrevirus)小麦矮缩病毒(Wheat DwarfVirus，WDV，中国农业科学院植物保护研究所王锡峰课题组惠赠)；甜菜曲顶病毒属(Curtovirus)甜菜曲顶病毒(Beet Curly Top Virus，BCTV，本实验室保藏)；菜豆金色花叶病毒属(Begomovirus)单组分病毒番茄黄化曲叶病毒(Tomato Yellow Leaf Curl Virus，TYLCV-SH2，中国农业科学院植物保护研究所周雪平课题组惠赠)和双组分病毒印度木薯花叶病毒(Indian Cassava Mosaic Virus，ICMV-SG，本实验室分离鉴定并保存，Gang Wanget al.，2014，Virus Genes)。本氏烟草的发病症状和病毒滴度两方面数据均表明瞬时表达IR-vTALE-AtKYP分子对双生病毒科病毒具有广谱抗性。
在小麦中瞬时表达步骤(1)构建的vTALE分子的优选组合IR-vTALE-AtKYP分子能够使小麦获得对小麦矮缩病毒的抗性。将分别含有IR-vTALE-MCS，IR-vTALE-AtKYP分子和WDV的农杆菌活化后，使其浓度达到OD600＝2.0，按vTALE分子和WDV体积比3∶1混合，穿刺接种小麦JN17(中国科学院遗传与发育生物学研究所李振生课题组惠赠)的胚及其周围位置，然后将接种的小麦种子种在土中，观察小麦的发病情况，发病症状和病毒滴度两方面结果均表明在小麦中瞬时表达IR-vTALE-AtKYP分子对WDV具有抗性。
在本氏烟中分别瞬时表达步骤(1)构建的vTALE分子和CRISPR/Cas9分子，接种甘蓝曲叶病毒后发现vTALE分子的优选组合RB-vTALE分子与CRISPR/Cas9分子相比能够使本氏烟草获得更好的抗性效果。将分别含有vTALE和CRISPR/Cas9分子的农杆菌注射本氏烟24h后，再接种甘蓝曲叶病毒的侵染性克隆，本氏烟的发病症状和病毒滴度两方面数据表明RB-vTALE分子具有比CRISPR/Cas9分子更好的抗CaLCuV的能力。
本发明人发现，在所选的植物物种中瞬时表达步骤(1)构建的vTALE分子能够赋予植物对双生病毒科病毒的广谱抗性。进一步地，本发明人验证了在植物中稳定表达步骤(1)构建的RB-vTALE分子的效果。
具体地，RB-vTALE-AtKYP分子转基因拟南芥植物具有良好的抗甘蓝曲叶病毒的效果。依照文献(Xiuren Zhang et al.，2006，Nature Protocol)报道的拟南芥转化方法，将含有步骤(1)构建的vTALE分子的优选组合RB-vTALE分子的农杆菌菌株分别转化Columbia生态型拟南芥野生型植物Col-0并收获T1代种子，用抗生素筛选T1代转基因种子从而成功获得转化RB-vTALE分子的T1代转基因植物株系。然后对转基因植物接种CaLCuV侵染性克隆，发病症状、病情指数和病毒滴度三方面数据表明RB-vTALE-AtKYP的转基因植物对甘蓝曲叶病毒具有很好的抗性。
RB-vTALE-AtKYP转基因拟南芥植物对甘蓝曲叶病毒(CaLCuV)的抗性效果可以稳定遗传给T3代，且这种抗性效果与AtKYP基因的表达水平呈正相关。对T3代RB-vTALE-AtKYP转基因拟南芥植物注射接种CaLCuV侵染性克隆，从发病症状、病情指数和病毒滴度三方面数据说明RB-vTALE-AtKYP的转基因植物对甘蓝曲叶病毒的抗性可以稳定遗传给下一代；对AtKYP基因的表达谱分析的结果表明RB-vTALE-AtKYP的转基因植物对甘蓝曲叶病毒的抗性与AtKYP基因的表达呈正相关。
T3代RB-vTALE-AtKYP转基因拟南芥植物中接种的甘蓝曲叶病毒基因组RB-vTALE靶点所在的核小体组蛋白和DNA均具有高水平的甲基化修饰，此外病毒相关基因的表达受到明显抑制。利用染色质免疫沉淀(Chromatin Immuno-precipitation，ChIP)技术检测T3代RB-vTALE-AtKYP转基因拟南芥植物中接种的甘蓝曲叶病毒基因组RB-vTALE靶点所在的核小体组蛋白的甲基化水平，发现组蛋白H3第9位赖氨酸甲基化水平明显提高；利用基于对甲基化敏感的限制性内切酶酶切消化的方法检测RB-vTALE靶点所在DNA的甲基化水平，发现DNA甲基化水平明显提高。
T4代RB-vTALE-AtKYP转基因拟南芥植物对甘蓝曲叶病毒(CaLCuV)具有很好的抗性效果，而且植物内源转座子Mu1和休眠相关基因的表达没有受RB-vTALE-AtKYP分子的影响。对T4代RB-vTALE-AtKYP转基因拟南芥植物注射接种CaLCuV侵染性克隆，从发病症状、病情指数和病毒滴度三方面数据说明RB-vTALE-AtKYP的转基因植物对甘蓝曲叶病毒的抗性可以稳定遗传到第四代；对植物内源转座子Mu1和休眠相关基因的表达谱分析的结果表明RB-vTALE-AtKYP的转基因植物中植物内源转座子Mu1和休眠相关基因的表达并没有明显变化，说明RB-vTALE-AtKYP分子并未脱靶作用于植物内源转座子Mu1和休眠相关基因上。
所述植物可以选自拟南芥、本氏烟草或小麦，但不限于此。
利用本发明的方法培育的广谱双生病毒抗性的植物可以抗多个双生病毒属的双生病毒，包括菜豆金色花叶病毒属中多种重要的病原甘蓝曲叶病毒(Cabbage Leaf CurlVirus，CaLCuV)，印度木薯花叶病毒(Indian Cassava Mosaic Virus，ICMV)，番茄黄化曲叶病毒(Tomato Yellow Leaf Curl Virus，TYLCV)；甜菜曲顶病毒属的甜菜曲顶病毒(BeetCurly Top Virus，BCTV)，或玉米线条病毒属的小麦矮缩病毒(Wheat Dwarf Virus，WDV)，但不限于这些病毒。
通过本发明的方法能使植物获得对双生病毒的广谱抗性，也可以使植物获得对某种双生病毒特异的、稳定的高抗效果，这两种抗性效果都是基于对双生病毒染色质进行表观遗传学修饰从而抑制病毒相关基因的复制或转录过程的策略。
综上所述，本发明提供下述技术方案：
1.一种改造的抗双生病毒的vTALE分子，其以植物病原细菌水稻黄单胞杆菌(Xanthomonas oryzae pv.oryzae)三型分泌系统效应因子AvrXa27基因序列为骨架序列，保留AvrXa27基因的N端和C端序列，然后对AvrXa27基因的DNA识别/结合结构域序列和功能输出结构域序列进行改造而获得，其中所述改造后的DNA识别/结合结构域序列靶向SEQ IDNO：1所示的IR-vTALE靶点序列或SEQ ID NO：2所示的RB-vTALE靶点序列，并且所述改造后的功能输出结构域序列选自由编码SEQ ID NOs：8-12所示的氨基酸序列的核苷酸序列组成的组中的任一种。
2.第1项所述的改造的抗双生病毒的vTALE分子，其中所述改造后的DNA识别/结合结构域序列靶向SEQ ID NO：1所示的IR-vTALE靶点序列。
3.第1项所述的改造的抗双生病毒的vTALE分子，其中所述改造后的DNA识别/结合结构域序列靶向SEQ ID NO：2所示的RB-vTALE靶点序列。
4.第1项所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列编码SEQ ID NO：8所示的AtKYP蛋白。
5.第1项所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列编码SEQ ID NO：9所示的AtHDAC19蛋白。
6.第1项所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列编码SEQ ID NO：10所示的AtMET1蛋白。
7.第1项所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列编码SEQ ID NO：11所示的AtSRDX蛋白。
8.第1项所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列编码SEQ ID NO：12所示的AtSUVH2蛋白。
9.第1项所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列是选自由SEQ ID NOs：3-7所示的核苷酸序列组成的组的任一种核苷酸序列。
10.第1项所述的改造的抗双生病毒的vTALE分子，其中所述vTALE分子的核苷酸序列如SEQ ID NOs：14-16和21-25所示。
11.第10项所述的改造的抗双生病毒的vTALE分子，其核苷酸序列如SEQ ID NO：14所示，命名为IR-vTALE-AtKYP。
12.第10项所述的改造的抗双生病毒的vTALE分子，其核苷酸序列如SEQ ID NO：21所示，命名为RB-vTALE-AtKYP。
13.一种培育具有稳定的广谱双生病毒抗性的植物的方法，所述方法包括下述步骤：
(1)vTALE分子的构建：所述vTALE分子以植物病原细菌水稻黄单胞杆菌(Xanthomonas oryzae pv.oryzae)三型分泌系统效应因子AvrXa27基因序列为骨架序列，保留AvrXa27基因的N端和C端序列，然后对AvrXa27基因的DNA识别/结合结构域序列和功能输出结构域序列进行改造而获得，其中所述改造后的DNA识别/结合结构域序列靶向SEQ IDNO：1所示的IR-vTALE靶点序列或SEQ ID NO：2所示的RB-vTALE靶点序列；并且所述改造后的功能输出结构域序列选自由编码SEQ ID NOs：8-12所示的氨基酸序列的核苷酸序列组成的组中的任一种；
(2)植物转化：将步骤(1)构建的vTALE分子，转化到所述植物中进行稳定表达。
14.第13项所述的方法，其中所培育的植物抗多个双生病毒属的双生病毒，包括菜豆金色花叶病毒属中多种重要的病原甘蓝曲叶病毒(Cabbage Leaf Curl Virus，CaLCuV)，印度木薯花叶病毒(Indian Cassava Mosaic Virus，ICMV)，番茄黄化曲叶病毒(TomatoYellow Leaf Curl Virus，TYLCV)；甜菜曲顶病毒属的甜菜曲顶病毒(Beet Curly TopVirus，BCTV)，或玉米线条病毒属的小麦矮缩病毒(Wheat Dwarf Virus，WDV)。
附图说明
从下面结合附图的详细描述中，本发明的上述特征和优点将更明显，其中：
图1显示vTALE分子构建示意图。
图1A.TALE分子结构示意图。现有技术中已有的一般TALE分子主要由两部分组成：DNA识别/结合结构域(Recognition/Binding domain)和激活结构域(Activation domain，AD)。其中DNA识别/结合结构域是由一段串联排列的重复序列组成，能够特异性识别并结合靶基因序列，每个重复序列由长度为33-35个氨基酸组成，除了第12和13位的氨基酸可变外，其他氨基酸序列高度保守，而重复序列可变的双氨基酸残基(Repeat variable di-residue，RVD)是TALE实现靶向识别特异DNA碱基的关键位点，不同的RVD能够相对特异地识别A、T、C、G四种碱基中的一种或多种，其中分别与这4种碱基相对应的最常见的4种RVD分别是NI＝A，NG＝T，HD＝C和NN＝G或A。激活结构域能够在TALE特异性识别并结合靶基因后激活靶基因的表达。
图1B.本发明设计的vTALE分子结构示意图。本发明设计的vTALE分子的DNA识别/结合结构域所结合的靶点序列选自下述中的任一种：针对双生病毒甘蓝曲叶病毒(Cabbageleaf curl virus，CaLCuV)基因组共同区(Common region，CR)序列设计的双生病毒科病毒高度保守的vTALE靶点Intergenetic Region-vTALE(IR-vTALE)和CaLCuV特异的含有复制相关蛋白(Replication associated protein，Rep)结合位点的vTALE靶点Rep Binding-vTALE(RB-vTALE)。此外，由于TALE蛋白所识别的靶点DNA序列必须以T开始，所以，虽然两个靶点序列都只含有16.5个串联排列的重复序列，但是都可以特异识别并结合18bp的靶点DNA序列。激活结构域被替换为分别融合表达拟南芥(Arabidopsis thaliana)不同的表观遗传学修饰相关效应因子(Epi-effector)，包括组蛋白甲基转移酶AtKYP，组蛋白去乙酰化酶AtHDAC19，DNA甲基转移酶AtMET1，转录抑制基序AtSRDX和虽同AtKYP同属一个家族，但是功能上不同的组蛋白甲基转移酶AtSUVH2。
图1C.双生病毒科不同属病毒的IR-vTALE靶点序列比对结果。Begomovirus，菜豆金色花叶病毒属；Turncurtovirus，芜菁曲顶病毒属；Curtovirus，甜菜曲顶病毒属；Topocuvirus，番茄伪曲顶病毒属；Mastrevirus，玉米线条病毒属。CaLCuV：甘蓝曲叶病毒，Cabbage Leaf Curl Virus；ICMV：印度木薯花叶病毒，Indian Cassava Mosaic Virus；TYLCV：番茄黄化曲叶病毒，Tomato Yellow Leaf Curl Virus；TCTV：芜菁曲顶病毒，TurnipCurly Top Virus；BCTV：甜菜曲顶病毒，Beet Curly Top Virus；TPCTV：番茄伪曲顶病毒，Tomato Pseudo-Curly Top Virus；WDV：小麦矮缩病毒，Wheat Dwarf Virus；ACMV：非洲木薯花叶病毒，African Cassava Mosaic Virus；EACMV：东非木薯花叶病毒，East AfricanCassava Mosaic Virus；EACMZV：东非木薯花叶病毒桑给巴尔分离株，East AfricanCassava Mosaic Zanzibar Virus；EACMKV：东非木薯花叶病毒肯尼亚分离株，EastAfrican Cassava Mosaic Kenya Virus。
图1D.本发明设计的vTALE和CRISPR/Cas9的靶点序列在CaLCuV基因组DNA-A链共同区中的位置示意图。两个vTALE靶点(IR-vTALE和RB-vTALE)及两个CRISPR/Cas9靶点(RB-sgRNA和TATA-sgRNA)均位于CaLCuV基因组DNA-A链的共同区内。蓝色框内序列为推测的CaLCuV由AC1编码的复制相关蛋白(Replication-associated protein，Rep)结合位点，其中下划线部分为保守序列；黄色框内序列为CaLCuV基因组中转录因子结合位点TATA box。蓝色曲线位置序列为双生病毒科病毒高度保守的IR-vTALE靶点，其转录方向同毒粒链方向，位于发夹环内(Haripin)；红色箭头位置序列为CaLCuV特异的RB-vTALE靶点，其转录方向同互补链方向，含有TATA box和推测的CaLCuV Rep蛋白结合位点序列。绿色箭头位置序列为CRISPR/Cas9的RB-sgRNA靶点，其转录方向同互补链方向，含有推测的CaLCuV Rep蛋白结合位点序列。紫色箭头位置序列为CRISPR/Cas9的TATA-sgRNA靶点，其转录方向同毒粒链方向，含有CaLCuV基因组中转录因子结合位点TATA box。
图2显示在本氏烟草中瞬时表达IR-vTALE-AtKYP分子对双生病毒科不同病毒的抗性效果。
图2A.显示在本氏烟草中瞬时表达空白载体对照IR-vTALE-MCS和IR-vTALE-AtKYP分子24h后接种双生病毒科不同病毒的发病症状。包括玉米线条病毒属(Mastrevirus)小麦矮缩病毒(Wheat Dwarf Virus，WDV)21dpi的症状；甜菜曲顶病毒属(Curtovirus)甜菜曲顶病毒(Beet Curly Top Virus，BCTV)5dpi的症状；菜豆金色花叶病毒属(Begomovirus)单组分病毒番茄黄化曲叶病毒(Tomato Yellow Leaf Curl Virus，TYLCV-SH2)10dpi的症状和双组分病毒印度木薯花叶病毒(Indian Cassava Mosaic Virus，ICMV-SG)7dpi的症状。Bar：1厘米。
图2B.显示在本氏烟草中瞬时表达空白载体对照IR-vTALE-MCS和IR-vTALE-AtKYP分子24h后接种双生病毒科不同病毒的病毒滴度。包括21dpi WDV，5dpi BCTV，10dpi TYLCV和7dpiICMV-SG的病毒滴度。
图3显示在小麦中瞬时表达IR-vTALE-AtKYP分子对小麦矮缩病毒(WDV)的抗性效果。
图3A.显示分别将IR-vTALE-MCS，IR-vTALE-AtKYP，RB-vTALE-MCS和RB-vTALE-AtKYP分子与WDV混合接种于小麦品种JN17种子胚及其周围位置10d的发病症状。Bar：1厘米。
图3B.显示分别将IR-vTALE-MCS和IR-vTALE-AtKYP分子与WDV混合接种于小麦品种JN17种子胚及其周围位置6个月后的发病症状。Bar：1厘米。
图3C.显示分别将IR-vTALE-MCS，IR-vTALE-AtKYP和RB-vTALE-AtKYP分子与WDV混合接种于小麦品种JN17种子胚及其周围位置10d后的病毒滴度。
图4显示在本氏烟中瞬时表达vTALE和CRISPR/Cas9分子对甘蓝曲叶病毒(CaLCuV)的抗性效果的比较。
图4A.显示注射MMA缓冲液的Mock对照。Bar：1厘米。
图4B-D.显示在本氏烟中瞬时表达载体对照sgRNA-vector(B)，TATA-sgRNA(C)和RB-sgRNA(D)分子24h后接种CaLCuV 14dpi的发病症状。Bar：1厘米。
图4E-I.显示在本氏烟中瞬时表达载体对照RB-vTALE-MCS(E)，RB-vTALE-AtKYP(F)，RB-vTALE-AtHDAC19(G)，RB-vTALE-AtMET1(H)和RB-vTALE-AtSRDX(I)分子24h后接种CaLCuV 14dpi的发病症状。Bar：1厘米。
图4J-L.显示在本氏烟中瞬时表达IR-vTALE-AtMET1(J)，IR-vTALE-AtHDAC19(K)和IR-vTALE-AtKYP(L)分子24h后接种CaLCuV 14dpi的发病症状。Bar：1厘米。
图4M.显示在本氏烟中瞬时表达vTALE和CRISPR/Cas9分子24h后接种CaLCuV14dpi的病毒滴度。
图5显示RB-vTALE-AtKYP转化拟南芥的植物对甘蓝曲叶病毒(CaLCuV)的抗性效果。
图5A.显示RB-vTALE-MCS，RB-vTALE-AtKYP，RB-vTALE-AtHDAC19，RB-vTALE-AtMET1和RB-vTALE-AtSUVH2转化拟南芥的T1代植物和Col-0对照接种CaLCuV两个月后的病情指数。
图5B-D.显示RB-vTALE-AtKYP(B)转化拟南芥的T2代植物和载体对照RB-vTALE-MCS(C和D)接种CaLCuV 1个月后的发病症状。
图5E.显示RB-vTALE-MCS和RB-vTALE-AtKYP(#1，#2，#4，#5，#6和#8)转化拟南芥的T2代植物和Col-0对照接种CaLCuV 1个月后的病毒滴度。
图6显示T3代RB-vTALE-AtKYP转基因拟南芥植物对甘蓝曲叶病毒(CaLCuV)的抗性效果及AtKYP基因的表达谱分析。
图6A.显示T3代RB-vTALE-MCS(#1)和RB-vTALE-AtKYP(#1和#4)转基因拟南芥植物接种CaLCuV 32天后的发病症状。
图6B.显示T3代RB-vTALE-MCS(#1)和RB-vTALE-AtKYP(#1，#2，#4，#8，#11，#13，#15和#18)转基因拟南芥和Col-0对照接种CaLCuV18天、25天及32天后的病情指数。
图6C.显示T3代RB-vTALE-MCS(#1)和RB-vTALE-AtKYP(#1和#4)转基因拟南芥接种CaLCuV 18天、25天及32天时的病毒滴度。
图6D.显示在T3代RB-vTALE-AtKYP(#1-3，#13-1，#15-1和#18-1)转基因拟南芥植物中AtKYP基因的表达谱分析。
图7显示T3代RB-vTALE-AtKYP转基因拟南芥植物中接种的甘蓝曲叶病毒基因组特定位置的DNA与组蛋白甲基化水平及病毒相关基因的表达谱的分析。
图7A.显示本发明中使用的CaLCuV基因组DNA-A链中核小体的分布情况，RB：RB-vTALE靶点，CR：共同区。
图7B.显示T3代RB-vTALE-AtKYP转基因植物对接种的CaLCuV基因组DNA-A链中#1核小体的组蛋白H3第9位和27位赖氨酸甲基化以及第14位赖氨酸乙酰化的影响。
图7C.显示T3代RB-vTALE-AtKYP转基因植物对接种的CaLCuV基因组DNA-A链中#5，#6，#8和#9核小体的组蛋白H3第9位和27位赖氨酸甲基化的影响。
图7D.显示T3代RB-vTALE-AtKYP转基因植物对接种的CaLCuV基因组DNA-A链中#1，#5，#6，#8和#9核小体的DNA甲基化的影响。
图7E.显示T3代RB-vTALE-AtKYP转基因植物对接种的CaLCuV的共同区(CommonRegion，CR)，互补链编码复制酶蛋白的基因ACl和互补链编码参与病毒复制或转录调控蛋白的基因AC4的表达谱的影响。
图8显示T4代RB-vTALE-AtKYP转基因拟南芥植物对甘蓝曲叶病毒(CaLCuV)的抗性效果、植物内源转座子Mu1和休眠相关基因的表达谱的影响。
图8A和8B.显示T4代RB-vTALE-AtKYP(B)转基因拟南芥植物和载体对照RB-vTALE-MCS(A)接种CaLCuV 21天时的发病症状。Bar：1厘米。
图8C.显示T4代RB-vTALE-AtKYP(#1-4和#1-12)转基因拟南芥植物和载体对照RB-vTALE-MCS(#1)接种CaLCuV 13天、21天及35天时的病情指数。
图8D.显示T4代RB-vTALE-AtKYP(#1-4和#1-12)转基因拟南芥植物和载体对照RB-vTALE-MCS(#1)接种CaLCuV 21天时的病毒滴度。
图8E.显示T4代RB-vTALE-AtKYP(#1-4和#1-12)和RB-vTALE-MCS(#1)转基因拟南芥叶片中植物内源转座子Mu1的表达谱分析。
图8F.显示T4代RB-vTALE-AtKYP(#1-12)和RB-vTALE-MCS(#1)转基因拟南芥种子中休眠相关基因(AtABI3、AtDOG1和AtNCED6)的表达谱分析。
图9显示IR-vTALE-MCS核苷酸序列(SEQ ID NO：13)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向IR-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列。
图10显示IR-vTALE-AtKYP核苷酸序列(SEQ ID NO：14)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向IR-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtKYP核苷酸序列。
图11显示IR-vTALE-AtHDAC19核苷酸序列(SEQ ID NO：15)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向IR-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtHDAC19核苷酸序列。
图12显示IR-vTALE-AtMET1核苷酸序列(SEQ ID NO：16)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向IR-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtMET1核苷酸序列。
图13显示IR-vTALE-AtKYP氨基酸序列(SEQ ID NO：17)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向IR-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtKYP氨基酸序列。
图14显示IR-vTALE-AtHDAC19氨基酸序列(SEQ ID NO：18)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向IR-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtHDAC19氨基酸序列。
图15显示IR-vTALE-AtMET1氨基酸序列(SEQ ID NO：19)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向IR-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtMET1氨基酸序列。
图16显示RB-vTALE-MCS核苷酸序列(SEQ ID NO：20)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向RB-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列。
图17显示RB-vTALE-AtKYP核苷酸序列(SEQ ID NO：21)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向RB-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtKYP核苷酸序列。
图18显示RB-vTALE-AtHDAC19核苷酸序列(SEQ ID NO：22)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向RB-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtHDAC19核苷酸序列。
图19显示RB-vTALE-AtMET1核苷酸序列(SEQ ID NO：23)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向RB-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtMET1核苷酸序列。
图20显示RB-vTALE-AtSRDX核苷酸序列(SEQ ID NO：24)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向RB-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtSRDX核苷酸序列。
图21显示RB-vTALE-AtSUVH2核苷酸序列(SEQ ID NO：25)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向RB-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtSUVH2核苷酸序列。
图22显示RB-vTALE-AtKYP氨基酸序列(SEQ ID NO：26)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向RB-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtKYP氨基酸序列。
图23显示RB-vTALE-AtHDAC19氨基酸序列(SEQ ID NO：27)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向RB-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtHDAC19氨基酸序列。
图24显示RB-vTALE-AtMET1氨基酸序列(SEQ ID NO：28)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向RB-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtMET1氨基酸序列。
图25显示RB-vTALE-AtSRDX氨基酸序列(SEQ ID NO：29)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向RB-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtSRDX氨基酸序列。
图26显示RB-vTALE-AtSUVH2氨基酸序列(SEQ ID NO：30)，其中阴影代表来源于Xoo的AvrXa27基因N端序列，下划线代表靶向RB-vTALE靶点的16.5个重复序列，斜体代表来源于Xoo的AvrXa27基因C端序列，粗体代表AtSUVH2氨基酸序列。
具体实施方式
下面参照具体的实施例进一步描述本发明，但是本领域技术人员应该理解，本发明并不限于这些具体的实施例。
实施例1.构建vTALE分子
转录激活效应子(Transcription activator-like effector，TALE)是一类来源于植物病原细菌黄单胞杆菌属(Xanthomonas spp.)三型分泌系统的效应因子，能够特异性识别并结合水稻的R基因从而激发水稻的抗病性(Keyu Gu et al.，2005，Nature)。但直到2009年，两个独立的实验室才解析了TALE特异性识别并结合靶基因的分子机制(Moscou etal.，2009，Science和Boch et al.，2009，Science)。如图1A所示，一般TALE蛋白主要由两部分组成：由串联排列的重复序列组成的DNA识别/结合结构域(Recognition/Bindingdomain)和激活结构域(Activation domain，AD)。其中组成DNA识别/结合结构域的串联重复序列由长度为33-35个氨基酸组成，除了第12和13位的氨基酸可变外，每个重复序列的氨基酸序列高度保守，而可变的第12和13位的氨基酸被称为重复序列可变的双氨基酸残基(Repeat variable di-residue，RVD)，是TALE特异性识别并结合DNA序列的关键位点，不同的RVD能够相对特异地识别A、T、C、G四种碱基中的一种或多种，其中分别与这4种碱基相对应的最常见的4种RVD分别是NI＝A，NG＝T，HD＝C和NN＝G或A。激活结构域能够在TALE特异性识别并结合靶基因后激活靶基因的表达。
本发明以双生病毒科病毒为研究对象，期望利用上述的TALE技术发展出一种提高植物对双生病毒抗性的新方法。我们的策略是人工设计全新的vTALE分子，该分子的DNA识别/结合结构域能够特异性识别并结合双生病毒基因组的一段序列，而激活结构域则被置换为具有抑制作用的表观遗传学修饰相关效应因子(Epi-effector)。因此当在植物中表达这种vTALE分子时，入侵的双生病毒就会被vTALE分子识别并特异性结合在病毒基因组靶点位置，然后融合表达的表观遗传学修饰相关效应因子就会对靶点位置的病毒染色质进行表观遗传学修饰，从而抑制双生病毒基因组DNA的复制和转录，达到提高植物对双生病毒抗性的目的(图1B)。
所有双生病毒基因组中均含有一段共同保守的非编码的基因间区序列——共同区(Common region，CR)，该区域富含病毒复制和转录的各种控制元件。如双生病毒科病毒番茄金色花叶病毒(Tomato Golden Mosaic Virus，TGMV)共同区第54-153bp的序列中含有复制相关蛋白(Replication associated protein，Rep)的结合位点5’-GGTAGTAAGGTAG-3’。Rep蛋白与病毒的复制相关，可以起始毒粒链DNA的合成，Rep蛋白结合位点是病毒特异性的，但具有一些重复的保守序列如TGMV的GGTAG序列，此外重复序列间的间隔序列也很重要如TGMV的间隔序列TAA。在Rep蛋白结合位点下游是转录因子结合位点TATA框(TATAbox)，它与病毒的转录有关。在TATA框下游，有段序列能形成发夹环的结构，该结构与病毒的复制有关，该环中存在保守的序列5’-TAATATTAC-3’(马修斯植物病毒学第八章第八节D3)。对双生病毒科不同属病毒含有发夹环序列的18bp DNA序列进行比对分析，发现该段序列在双生病毒科内高度保守(图1C)，因此我们将该段18bp DNA序列作为vTALE分子的Intergenic Region-vTALE(IR-vTALE)靶点，希望通过序列高度保守的IR-vTALE靶点获得对双生病毒科病毒的广谱抗性。另外我们分析了CaLCuV基因组共同区第91-192bp的序列，结果如图1D所示，在CaLCuV共同区中存在可能的Rep蛋白结合位点5’-GGTGTCTTGGTGT-3’，TATA框和发夹环结构，然后我们将CaLCuV含有TATA框和部分Rep蛋白结合位点的18bp DNA序列作为vTALE分子的Rep Binding-vTALE(RB-vTALE)靶点，希望通过病毒特异性的RB-vTALE靶点获得对CaLCuV更特异性的抗性(图1B和1D)。
与IR-vTALE和RB-vTALE靶点融合表达的表观遗传学修饰相关效应因子均来自于模式植物拟南芥(Arabidopsis thaliana)，包括组蛋白甲基转移酶AtKYP，组蛋白去乙酰化酶AtHDAC19，DNA甲基转移酶AtMET1，转录抑制基序AtSRDX和虽同AtKYP同属一个家族，但是功能上不同的组蛋白甲基转移酶AtSUVH2。
参照文献(Feng Zhang et al.，2011，Nature biotechnology)报道的基于层级连接的策略构建TALE蛋白DNA识别/结合结构域的串联排列重复序列的方法，构建3个IR-vTALE-Epi-effector分子、5个RB-vTALE-Epi-effector分子及其对应的载体对照分子IR-vTALE-MCS和RB-vTALE-MCS，详细信息见表1。
将构建好的10个vTALE分子分别转化农杆菌菌株C58C1的感受态细胞(购自北京博迈德生物技术有限公司)，利用含有相应抗生素的LB固体平板筛选阳性克隆。
实施例2.在本氏烟草中瞬时表达IR-vTALE-AtKYP分子对双生病毒科病毒具有广谱抗性。
将双子叶植物本氏烟草(Nicotiana benthamiana)种子均匀播种于灭菌的营养土表面，浇水并覆盖塑料薄膜以充分保湿，置于25℃植物培养箱中14小时光照/10小时黑暗培养10天左右，然后将萌发的小苗单株移栽到含有灭菌营养土的营养钵中，再生长3周左右即可用于实验。
将含有IR-vTALE-MCS(IR-vTALE分子空白载体对照)和IR-vTALE-AtKYP的农杆菌接种于含有相应抗生素的LB液体培养基中活化培养24-36小时(抗生素见表1)，然后取约1ml新鲜菌液接种于含有相应抗生素、MES(10mM)和AS(40μM)的20ml LB液体培养基中培养16-24小时，待OD600＝1.0-1.5之间时，离心收集菌体，再用MMA缓冲液重悬(10mM MES，10mMMgCl2，200μM AS)，调OD600＝1.0，静止2h后用1ml注射器接种本氏烟草叶片背面，使整片叶子充分浸润菌液。注射后的本氏烟在25℃，14h光照/10h黑暗条件下的培养箱中生长24小时后，使用相同方法按照表1的抗生素种类及浓度分别重新接种双生病毒科不同属病毒的侵染性克隆，包括玉米线条病毒属(Mastrevirus)小麦矮缩病毒(Wheat Dwarf Virus，WDV，中国农业科学院植物保护研究所王锡峰课题组惠赠)；甜菜曲顶病毒属(Curtovirus)甜菜曲顶病毒(Beet Curly Top Virus，BCTV，本实验室保藏)；菜豆金色花叶病毒属(Begomovirus)单组分病毒番茄黄化曲叶病毒(Tomato Yellow Leaf Curl Virus，TYLCV-SH2，中国农业科学院植物保护研究所周雪平课题组惠赠)和双组分病毒印度木薯花叶病毒(Indian Cassava Mosaic Virus，ICMV-SG，本实验室分离鉴定并保存，Gang Wang et al.，2014，Virus Genes)。接种后的本氏烟草置于25℃，14h光照/10h黑暗条件下的培养箱中生长，观察发病情况。
BCTV，ICMV-SG，TYLCV和WDV分别于接种后的第5天，第7天，第10天和第21天在表达空白载体对照IR-vTALE-MCS分子的本氏烟中出现明显的发病症状，其中接种WDV的叶片明显增厚和皱缩，接种BCTV的叶片出现半透明状沿叶脉扩展的症状，接种TYLCV的叶片出现明显的增厚、皱缩和向下扭曲的症状，接种ICMV-SG的叶片出现严重的增厚、皱缩和向下扭曲的症状，而表达IR-vTALE-AtKYP分子的本氏烟中未见明显的发病症状(图2A)。
为了进一步验证观察到的结果，我们取发病的样品进行病毒滴度的测定。每个处理至少取四个生物学重复，每一个样品100mg，使用植物基因组DNA提取试剂盒(购自Qiagen公司)提取基因组DNA。取2μl DNA为模板，0.4μl 10μm的引物，10μl SYBR Green(TOYOBO)，然后用ddH2O补足至20μl。实时PCR程序为预变性95℃10min，变性95℃15s，退火58℃30s，延伸72℃30s，40个循环，补平延伸5min。所有样品都做三次技术重复。NbEF1a为内参基因(引物见表2)，进行实时PCR分析。
如图2B所示，BCTV，ICMV-SG，TYLCV和WDV在表达空白载体对照IR-vTALE-MCS分子的本氏烟中的病毒滴度明显高于表达IR-vTALE-AtKYP分子的本氏烟，其中WDV在瞬时表达IR-vTALE-MCS分子的本氏烟中的病毒滴度是瞬时表达IR-vTALE-AtKYP分子的本氏烟的3倍多，BCTV在瞬时表达IR-vTALE-MCS分子的本氏烟中的病毒滴度是瞬时表达IR-vTALE-AtKYP分子的本氏烟的1倍多，TYLCV在瞬时表达IR-vTALE-MCS分子的本氏烟中的病毒滴度是瞬时表达IR-vTALE-AtKYP分子的本氏烟的1倍多，ICMV-SG在瞬时表达IR-vTALE-MCS分子的本氏烟中的病毒滴度是瞬时表达IR-vTALE-AtKYP分子的本氏烟的约500000倍，这与我们观察到的发病结果相吻合，说明瞬时表达IR-vTALE-AtKYP分子的本氏烟表现出抗多种双生病毒侵染的能力。
结论：针对双生病毒CaLCuV基因组共同区序列设计的双生病毒科病毒序列高度保守的vTALE靶点IR-vTALE融合表达拟南芥组蛋白甲基转移酶KYP的vTALE分子IR-vTALE-AtKYP在双子叶植物本氏烟草中瞬时表达24小时后即能够增强本氏烟对多种双生病毒的抗性能力，也就是说IR-vTALE-AtKYP分子对双生病毒科病毒具有广谱抗性的效果。
表2.本发明中病毒滴度实验所用到的引物
实施例3.在小麦中瞬时表达IR-vTALE-AtKYP分子对小麦矮缩病毒具有抗性。
将分别含有IR-vTALE-MCS(IR-vTALE空白载体对照)，IR-vTALE-AtKYP，RB-vTALE-MCS(RB-vTALE空白载体对照)，RB-vTALE-AtKYP和WDV侵染性克隆的农杆菌接种于含有相应抗生素的3ml LB液体培养基中(抗生素见表1)，活化培养至OD600＝2.0，离心收集菌体，再用MMA缓冲液浓缩至0.75ml(10mM MES，10mM MgCl2，200μM AS)，静止2h后，分别将vTALE分子与WDV侵染性克隆按3∶1的比例混匀，然后用昆虫针蘸取混好的菌液，穿刺接种于灭菌水浸泡过夜的小麦(Triticum aestivum Linn)品种JN17(中国科学院遗传与发育生物学研究所李振生课题组惠赠)的胚及其周围位置，每个种子从不同角度接种五次，然后将接种的种子置于不含抗生素的LB固体平板上28℃过夜培养，第二天将其均匀播种于灭菌的营养土中，25℃植物培养箱中18小时光照/6小时黑暗培养2-4周，观察发病情况。
在vTALE和WDV混合接种10天后，WDV分别与IR-vTALE-MCS(IR-vTALE载体对照)，RB-vTALE-MCS(RB-vTALE载体对照)和RB-vTALE-AtKYP混合接种的小麦与空白对照Mock植物相比植株明显矮小，生长状态不佳；而WDV与IR-vTALE-AtKYP接种的小麦表现出比其他vTALE分子接种的小麦更健康的表型，植株明显高大、健壮，生长状态更接近于Mock对照(图3A)。
为了进一步验证观察到的结果，我们按照实施例2中所述的病毒滴度测定方法以小麦actin为内参基因(引物见表2)对混合接种vTALE和WDV的小麦进行病毒滴度的测定。
如图3C所示，WDV在与IR-vTALE-MCS(IR-vTALE载体对照)混合接种的小麦中的病毒滴度是与IR-vTALE-AtKYP混合接种的小麦的约4倍，而WDV在与RB-vTALE-AtKYP混合接种的小麦中的病毒滴度是与IR-vTALE-MCS(IR-vTALE载体对照)混合接种的小麦的约1.5倍，这与我们观察到的发病结果相吻合，说明瞬时表达IR-vTALE-AtKYP分子的小麦表现出高抗WDV侵染的能力，而RB-vTALE-AtKYP分子没有抵抗WDV侵染的能力。
为了更好地检验瞬时表达IR-vTALE-AtKYP分子的小麦抗小麦矮缩病毒(WheatDwarf Virus，WDV)侵染的效果，我们将vTALE分子和WDV混合接种的小麦移栽到大钵中，在适宜小麦生长的季节将其置于室外自然条件下生长，在注射接种后的6个月，我们发现WDV与IR-vTALE-AtKYP混合接种的小麦植株明显高大、健壮、能够正常抽穗，生长状态更接近于Mock对照；而WDV与IR-vTALE-MCS(IR-vTALE载体对照)混合接种的小麦植株矮小、几乎绝产(图3B)。说明瞬时表达IR-vTALE-AtKYP分子的小麦对WDV的侵染具有高抗且持久的效果。
结论：针对双生病毒CaLCuV基因组共同区序列设计的双生病毒科病毒高度保守的TALE靶点IR-vTALE融合表达拟南芥组蛋白甲基转移酶KYP的vTALE分子IR-vTALE-AtKYP在单子叶作物小麦中瞬时表达后能够增强小麦对WDV的抗性能力，这一研究结果在培育高抗植物双生病毒的小麦品种方面具有很好的应用前景。而CaLCuV特异的TALE靶点RB-vTALE融合表达拟南芥组蛋白甲基转移酶KYP的vTALE分子RB-vTALE-AtKYP没有抵抗WDV侵染的能力。
实施例4.在本氏烟中瞬时表达vTALE和CRISPR/Cas9分子对甘蓝曲叶病毒的抗性效果比较。
首先，我们针对CaLCuV基因组DNA-A链共同区序列设计包括TATA-sgRNA(A2)和RB-sgRNA(A1)在内的8个CRISPR/Cas9靶点(引物序列见表3)，然后使用annealing buffer(购自碧云天生物技术有限公司)将每个靶点的正、反向引物退火形成互补的双链DNA分子，最后将annealing产物与被BsaI消化过的载体pHSN401(中国科学院遗传与发育生物学研究所高彩霞课题组惠赠)进行连接反应，4小时后将连接产物转化大肠杆菌DH5α感受态(购自北京博迈德生物技术有限公司)，复苏1小时后涂布于含有卡那霉素(50mg/L)的LB固体平板上筛选阳性克隆，挑取阳性克隆提取质粒送测，测序结果比对正确的克隆转化农杆菌C58C1感受态(购自北京博迈德生物技术有限公司)，利用含有卡那霉素(50mg/L)，利福平(20mg/L)和四环素(10mg/L)的固体LB平板筛选阳性克隆。
其次，按照实施例2中所述的方法对双子叶植物本氏烟进行vTALE和CRISPR/Cas9分子的瞬时表达，24h后再重新注射接种CaLCuV侵染性克隆(本实验室保存)，观察发病情况。
接种CaLCuV侵染性克隆14天时我们发现与Mock植物相比(图4A)，瞬时表达CRISPR/Cas9载体对照sgRNA-vector的本氏烟叶片上有明显的CaLCuV感染的症状，包括叶片上呈现褪绿斑点，叶片明显增厚并向下卷曲(图4B)；而除了瞬时表达RB-sgRNA(A1)靶点的本氏烟叶片上未见明显的CaLCuV感染的症状外(图4D)，瞬时表达其他CRISPR/Cas9靶点的本氏烟均具有与载体对照类似的明显的CaLCuV感染的症状(图4C，更具体的数据未提供)，说明我们设计的8个CRISPR/Cas9靶点中只有RB-sgRNA(A1)靶点具有相对比较好的抗CaLCuV侵染的能力。另一方面，瞬时表达vTALE载体对照RB-vTALE-MCS的本氏烟叶片上有明显的与sgRNA-vector类似的CaLCuV感染的症状(图4E)，而瞬时表达RB-vTALE-Epi-effector【RB-vTALE-AtKYP(图4F)，RB-vTALE-AtHDAC19(图4G)，RB-vTALE-AtMET1(图4H)和RB-vTALE-AtSRDX(图4I)】的本氏烟叶片上未见明显的CaLCuV感染的症状，与瞬时表达CRISPR/Cas9靶点RB-sgRNA(A1)的本氏烟类似；而IR-vTALE-Epi-effector中除了瞬时表达IR-vTALE-AtKYP的本氏烟叶片上有比较微弱的CaLCuV感染的症状外(图4L)，瞬时表达IR-vTALE-AtMET1(图4J)和IR-vTALE-AtHDAC19(图4K)的本氏烟叶片上有明显的CaLCuV感染的症状，其发病程度与载体对照RB-vTALE-MCS类似(图4E)，说明从总体来讲RB-vTALE分子具有比CRISPR/Cas9分子更强的抗CaLCuV侵染的效果。
为了进一步验证观察到的结果，我们按照实施例2中所述的病毒滴度测定方法以NbEF1a为内参基因(引物见表2)检测分别瞬时表达vTALE和CRISPR-Cas9分子的本氏烟中CaLCuV的病毒滴度。
如图4M所示，CaLCuV在本氏烟叶片中的病毒滴度由低到高依次为瞬时表达RB-vTALE-Epi-effector(RB-vTALE-AtMET1，RB-vTALE-AtKYP，RB-vTALE-AtSRDX和RB-vTALE-AtHDAC19)的本氏烟叶片，平均病毒滴度不到载体对照的万分之一；其次是瞬时表达CRISPR/Cas9靶点RB-sgRNA(A1)的本氏烟叶片，病毒滴度比载体对照的万分之一略高；最后是瞬时表达IR-vTALE-AtKYP的本氏烟叶片，病毒滴度约为载体对照的千分之一；而瞬时表达其他vTALE和CRISPR/Cas9分子的本氏烟中CaLCuV的病毒滴度与瞬时表达相应的载体对照相类似。说明RB-vTALE-Epi-effectors分子对CaLCuV的侵染具有比CRISPR/Cas9分子更好的抗性效果，此外IR-vTALE-AtKYP对CaLCuV的侵染也具有较好的抗性效果，这一结果与我们观察到的发病情况一致。
结论：CaLCuV特异性的靶点RB-vTALE融合表达表观遗传学修饰相关效应因子的vTALE分子对CaLCuV的侵染具有比CRISPR/Cas9分子更好的抗性效果，此外对双生病毒科病毒具有广谱抗性的IR-vTALE靶点融合表达拟南芥组蛋白甲基转移酶KYP的vTALE分子对CaLCuV的侵染也具有较好的抗性效果。
表3.本发明中针对CaLCuV设计的CRISPR/Cas9靶点序列引物
引物名称 引物序列(5’-3’) 目的
RB-sgRNA(A1)F ATTGACCAAGACACCAAAAACAAT CRISPR/Cas9
RB-sgRNA(A1)R AAACATTGTTTTTGGTGTCTTGGT CRISPR/Cas9
TATA-sgRNA(A2)F ATTGAATATATACTAAAAGCCTCT CRISPR/Cas9
TATA-sgRNA(A2)R AAACAGAGGCTTTTAGTATATATT CRISPR/Cas9
A3F ATTGAAAGCCTCTGGGGAACACCA CRISPR/Cas9
A3R AAACTGGTGTTCCCCAGAGGCTTT CRISPR/Cas9
A4F ATTGGGGAACACCAGGGGCAAAAG CRISPR/Cas9
A4R AAACCTTTTGCCCCTGGTGTTCCC CRISPR/Cas9
A5F ATTGGCCATCCGCAATAATATTAC CRISPR/Cas9
A5R AAACGTAATATTATTGCGGATGGC CRISPR/Cas9
A6F ATTGCGGATGGCCGCTTTTGCCCC CRISPR/Cas9
A6R AAACGGGGCAAAAGCGGCCATCCG CRISPR/Cas9
A7F ATTGTCCGGTAATATTATTGCGGA CRISPR/Cas9
A7R AAACTCCGCAATAATATTACCGGA CRISPR/Cas9
A8F ATTGCCGGATGGCCGCGATTTTTT CRISPR/Cas9
A8R AAACAAAAAATCGCGGCCATCCGG CRISPR/Cas9
实施例5.RB-vTALE-AtKYP转化拟南芥的植物对甘蓝曲叶病毒的抗性效果分析。
由于在本氏烟中瞬时表达不同的RB-vTALE-Epi-effector对CaLCuV的侵染均具有很好的抗性效果，因此我们想检测稳定表达不同的RB-vTALE-Epi-effector对CaLCuV的抗性效果。依照文献(Xiuren Zhang et al.，2006，Nature Protocol)报道的拟南芥转化方法分别将含有不同RB-vTALE-Epi-effector分子的农杆菌菌株C58C1转化野生型拟南芥Col-0，再对T1代种子进行转化子筛选，从而成功获得RB-vTALE-MCS，RB-vTALE-AtKYP，RB-vTALE-AtHDAC19，RB-vTALE-AtMET1和RB-vTALE-AtSUVH2转基因拟南芥植物。
然后，按照实施例2中所述的注射接种本氏烟草的方法将CaLCuV侵染性克隆接种到4周左右的RB-vTALE-MCS，RB-vTALE-AtKYP，RB-vTALE-AtHDAC19，RB-vTALE-AtMET1和RB-vTALE-AtSUVH2转基因拟南芥的轮作叶片上，同时接种Col-0做对照，观察发病情况。
接种CalCuV 2个月后，我们将拟南芥发病情况按由轻到重的顺序划分为6个等级(0-5级)，利用病情指数这一指标初步统计每个vTALE分子转基因植物对CaLCuV的抗性效果，结果如图5A所示，T1代RB-vTALE-AtHDAC19，RB-vTALE-AtMET1和RB-vTALE-AtKYP转基因拟南芥都表现出抗CaLCuV侵染的效果，但是RB-vTALE-AtSUVH2转基因植物表现出与载体对照和Col-0相一致的表型，说明AtSUVH2虽为拟南芥的另一个组蛋白甲基转移酶，同AtKYP同属一个家族，但是功能上不同，对CaLCuV的侵染没有抗性效果。
待种子成熟后，挑选载体对照RB-vTALE-MCS和抗性比较好的RB-vTALE-AtKYP转基因株系的种子萌发T2代植株，按照实施例2中所述的注射接种本氏烟草的方法将CaLCuV侵染性克隆接种到4周左右的转基因拟南芥轮作叶上，观察发病情况。
接种CalCuV 1个月后，T2代RB-vTALE-AtKYP转基因株系表现出高抗表型，植株生长健康，正常开花，无明显的叶片斑驳或褪绿的症状(图5B)，而T2代载体对照RB-vTALE-MCS转基因株系表现出易感表型，包括植株矮小，花絮扭曲，花叶等症状(图5C和D)。
为了进一步验证观察到的结果，我们按照实施例2中所述的病毒滴度测定方法以拟南芥Tubulin2为内参基因(引物见表2)对接种CalCuV的转基因拟南芥进行病毒滴度的测定。
如图5E所示，病毒滴度分析的结果表明与载体对照RB-vTALE-MCS转基因拟南芥植物相比RB-vTALE-AtKYP转基因拟南芥植物株系(#1，#4，#8)对CaLCuV具有高抗的表型，其中RB-vTALE-AtKYP转基因拟南芥植物株系#1的病毒滴度约为载体对照植物的1/14，#4和#8的病毒滴度约为载体对照植物的1/3，这一结果与观察到的发病症状的结果相一致。
结论：CaLCuV特异的靶点RB-vTALE融合表达拟南芥组蛋白甲基转移酶KYP的RB-vTALE-AtKYP分子在双子叶植物拟南芥中稳定表达后对CaLCuV的侵染具有很好的抗性效果。
实施例6.T3代RB-vTALE-AtKYP转基因拟南芥植物对甘蓝曲叶病毒(CaLCuV)的抗性效果及AtKYP基因的表达谱分析。
待T3代转基因植物种子成熟后，挑选载体对照RB-vTALE-MCS和抗性比较好的RB-vTALE-AtKYP转基因株系的种子萌发T3代植株，按照实施例2中所述的注射接种本氏烟草的方法将CaLCuV侵染性克隆接种到4周左右的转基因拟南芥轮作叶上，观察发病情况。
接种CalCuV 32天后，T3代RB-vTALE-AtKYP转基因株系(#1和#4)表现出高抗表型，植株生长健康，正常开花，无明显的叶片斑驳或褪绿的症状，而T3代载体对照RB-vTALE-MCS#1转基因株系表现出易感表型，包括植株矮小，花絮扭曲，花叶等症状，如图6A所示，其中左上角为RB-vTALE-MCS#1转基因株系发病症状的俯视放大图。
利用病情指数这一指标按发病早、中和晚期三个时间点统计的RB-vTALE-AtKYP转基因株系对CaLCuV的抗性效果如图6B所示，RB-vTALE-AtKYP转基因株系(#1，#4，#8和#18)在发病的早、中和晚期均对CaLCuV具有很好的抗性效果，说明RB-vTALE-AtKYP转基因拟南芥对CaLCuV的抗性效果可以稳定遗传给下一代。
为了进一步验证观察到的结果，我们按照实施例2中所述的病毒滴度测定方法以拟南芥Tubulin2为内参基因(引物见表2)对接种CalCuV的转基因拟南芥进行病毒滴度的测定。
如图6C所示，病毒滴度分析的结果表明RB-vTALE-AtKYP转基因植物株系(#1和#4)在发病的早、中和晚期对CaLCuV都具有高抗的效果，发病中期的抗性效果尤为显著，这一结果与观察到的发病症状及病情指数调查的结果相一致。
为了检测不同RB-vTALE-AtKYP转基因植物株系中AtKYP基因的表达水平，我们选取高抗株系#1-3和#18-1及易感株系#13-1和#15-1为待测样品。首先按AtKYP基因核苷酸序列为模板设计扩增引物(见表4，AtKYP-F和AtKYP-R)，然后每种植物至少三个生物学重复，每一个样品取100mg，使用RNA提取试剂盒(购自Qiagen)提取总RNA并对基因组DNA进行消化处理，Nanodrop定量后取2μg总RNA为模板，MMLV反转录酶合成cDNA。取2μl cDNA为模板，0.4μl 10μm的引物，10μl SYBR Green(TOYOBO)，然后用ddH2O补足至20μl。实时PCR程序为预变性95℃10min，变性95℃15s，退火58℃30s，延伸72℃30s，40个循环，补平延伸5min。所有样品都做三次技术重复。AtActin2为内参基因(引物见表4，AtActin2-F和AtActin2-R)，进行实时PCR分析。如图6D所示，RB-vTALE-AtKYP转基因植物高抗株系#1-3和#18-1中AtKYP基因的表达水平是易感株系#13-1和#15-1的约7倍。
结论：CaLCuV特异的靶点RB-vTALE融合表达拟南芥组蛋白甲基转移酶KYP的RB-vTALE-AtKYP分子在双子叶植物拟南芥中稳定表达后对CaLCuV的高抗表型能够稳定遗传给下一代，且与AtKYP基因表达水平呈正相关。
表4.本发明中用于基因表达分析的引物
实施例7.T3代RB-vTALE-AtKYP转基因拟南芥植物中接种的甘蓝曲叶病毒基因组特定位置的组蛋白与DNA甲基化水平及病毒相关基因的表达谱的分析
由于AtKYP是来源于拟南芥的组蛋白H3第9位赖氨酸甲基化转移酶基因，而前期的研究结果已经证明RB-vTALE-AtKYP转基因拟南芥植物对CaLCuV的高抗表型与AtKYP基因表达水平呈正相关，且能够稳定遗传给下一代。这就暗示我们RB-vTALE-AtKYP转基因拟南芥植物对CaLCuV的高抗表型可能是通过AtKYP甲基化修饰CaLCuV基因组RB-vTALE靶点所在的核小体组蛋白，进而引起DNA的甲基化修饰，使得CaLCuV基因组不能正常复制和转录，最终使得RB-vTALE-AtKYP转基因拟南芥植物获得对CaLCuV的抗性。为了验证这一推论，首先我们利用染色质免疫沉淀(Chromatin Immuno-precipitation，ChIP)技术来检测CaLCuV基因组RB-vTALE靶点所在核小体即#1核小体组蛋白的甲基化水平(图7A)。
根据文献(In-Cheol Jang et al.，2011，The plant cell)报道的方法，首先取相同叶位的发病叶片3g，每个处理至少三个生物学重复，将样品浸润在终浓度为1％甲醛中，抽真空10min使染色质结合蛋白与DNA交联，然后加入终浓度为0.125M的甘氨酸抽真空5min以终止交联反应，将样品用灭菌水冲洗三遍后，用液氮速冻并磨成粉末，再用一系列抽提缓冲液处理后，将收集的染色质经过超声波打断，再用识别特异组蛋白修饰的抗体(anti-H3K9me2，anti-H3K27me3和anti-H3K14ac均购自Millipore公司)将核小体沉淀下来，缠绕在核小体上的DNA也随之沉淀，解交联用蛋白酶K除去蛋白质后用PCR纯化试剂盒(购自Qiagen公司)纯化出DNA，再设计基因组特异位点的引物用定量PCR的方法检测DNA的量(引物序列见表5)，DNA的量的多少就可以反映出该位点对应的组蛋白修饰的相对富集程度。
结果如图7B所示，RB-vTALE-AtKYP转基因拟南芥植物株系#1-12中CaLCuV基因组的#1核小体的组蛋白H3第9位赖氨酸的甲基化水平与载体对照植物中的相比提高10倍，说明RB-vTALE-AtKYP转基因拟南芥植物株系#1-12中RB-vTALE-AtKYP分子结合于CaLCuV基因组的RB-vTALE靶点，然后AtKYP对靶点所在的#1核小体的组蛋白H3第9位赖氨酸进行甲基化修饰。但是与此同时我们发现CaLCuV基因组的#1核小体所在的组蛋白H3第27位赖氨酸的甲基化水平与载体对照植物中的相比也提高5倍多，这可能也是由AtKYP引起的，但是具体的机制我们还不清楚。此外我们发现CaLCuV基因组的#1核小体所在的组蛋白H3第14位赖氨酸的乙酰化水平与载体对照植物中的相比没有明显变化，说明RB-vTALE-AtKYP转基因拟南芥植物株系#1-12中RB-vTALE-AtKYP分子对CaLCuV基因组的#1核小体的组蛋白乙酰化没有作用，而只特异性地作用于甲基化修饰上。
既然RB-vTALE-AtKYP转基因拟南芥植物中RB-vTALE-AtKYP分子能够甲基化修饰CaLCuV基因组的RB-vTALE靶点所在的#1核小体的组蛋白，那么RB-vTALE-AtKYP分子是否对CaLCuV基因组中其他核小体的组蛋白也具有甲基化修饰的作用呢？带着这个问题我们选择了与CaLCuV基因组RB-vTALE靶点平面距离较近的#8和#9核小体及平面距离较远的#5和#6核小体(图7A)来检测它们的组蛋白的甲基化修饰水平。
结果如图7C所示，RB-vTALE-AtKYP转基因拟南芥植物中CaLCuV基因组的#9核小体的组蛋白H3第9位赖氨酸的甲基化水平与载体对照植物中的相比提高16倍，而#5、#6和#8核小体的组蛋白H3第9位赖氨酸的甲基化水平与载体对照植物中的相比并没有明显的变化。说明RB-vTALE-AtKYP转基因拟南芥植物中RB-vTALE-AtKYP分子能够甲基化修饰与其识别位点RB-vTALE靶点平面距离最近的#9核小体的组蛋白，但不能甲基化修饰与RB-vTALE靶点平面距离较远的#5、#6和#8核小体的组蛋白。此外我们发现RB-vTALE-AtKYP转基因拟南芥植物中CaLCuV基因组的#5核小体的组蛋白H3第27位赖氨酸的甲基化水平与载体对照植物中的相比提高12倍，而#6、#8和#9核小体的组蛋白H3第27位赖氨酸的甲基化水平并没有明显的变化，这可能是由AtKYP引起的，虽然#5核小体与RB-vTALE靶点所在的#1核小体平面距离最远，但是CaLCuV基因组DNA在复制过程中会形成双链结构的复制中间体，进而招募寄主植物的组蛋白形成类似植物染色体结构的minichromosome，这种结构可能会拉近二者的空间距离，从而使得AtKYP影响#5核小体的组蛋白H3第27位赖氨酸甲基化修饰的水平，但是具体原因我们还不能很好地解释。
据文献(Jiamu Du et al.，2014，Molecular cell)报道，拟南芥核小体组蛋白H3第9位赖氨酸在被KYP修饰带有甲基化Marker后，就会招募CMT3(DNA甲基转移酶CHROMOMETHYLASE3)对其DNA进行甲基化修饰，带有甲基化Marker的DNA又会招募KYP对其所在核小体的组蛋白或与其相毗邻的核小体组蛋白H3第9位赖氨酸进行甲基化修饰，二者作为分子伴侣通过这样的方式共同控制核小体组蛋白H3第9位赖氨酸及其DNA的甲基化修饰。前面结果显示RB-vTALE-AtKYP转基因拟南芥植物中CaLCuV基因组的#1和#9核小体的组蛋白H3第9位赖氨酸的甲基化水平与载体对照植物中的相比有明显提高，因此我们采用以下方法来检测RB-vTALE-AtKYP转基因拟南芥植物中CaLCuV基因组RB-vTALE靶点所在#1核小体及其他核小体(#5，#6，#8和#9)内DNA甲基化水平。
取相同叶位的发病叶片，每一个样品100mg，每个处理至少三个生物学重复，使用基因组DNA提取试剂盒(购自Qiagen)进行高纯度基因组的提取；每个样品取400ng的基因组DNA，分装两管，向处理管中加入3U甲基化敏感性限制性内切酶HpaII，相应的对照管中加入等体积的ddH2O，每管中加入3ul酶切缓冲液后用ddH2O定容至30ul，37℃单酶切消化5h后，加热至80℃失活20min；向酶切体系中加入50ul ddH2O稀释，取2ul作为模板，以AtTubulin2为内参基因按照实施例2中所述的测定病毒滴度的反应体系及程序进行Real-time PCR分析，所用的引物序列见表5。
结果如图7D所示，RB-vTALE-AtKYP转基因拟南芥植物中CaLCuV基因组RB-vTALE靶点所在#1核小体及与其平面距离最近的#9核小体内的DNA甲基化水平与载体对照植物中的相比有所提升，而与RB-TALE靶点平面距离较远的#5，#6和#8核小体内的DNA甲基化水平与载体对照植物中的相比并没有明显的差异。说明RB-vTALE-AtKYP转基因拟南芥植物中RB-vTALE-AtKYP分子能够对其靶点所在及其毗邻的核小体组蛋白进行甲基化修饰，进而招募CMT3使其对DNA进行甲基化修饰，但是对于其他核小体(#5，#6和#8核小体)的DNA没有作用。
由于RB-vTALE靶点所在的#1核小体DNA序列位于CaLCuV的共同区，CaLCuV的复制及转录起始相关酶均结合在共同区内特异位点上，而我们的数据表明RB-vTALE-AtKYP转基因拟南芥植物中CaLCuV基因组#1核小体DNA甲基化水平有所提高，因此想要检测一下病毒相关基因的表达水平是否受到影响。我们按照实施例6中所述的检测目的基因表达水平的方法对CaLCuV的共同区(Common Region，CR)，互补链编码复制酶蛋白的基因AC1和互补链编码参与病毒复制或转录调控蛋白的基因AC4的表达水平进行检测。
结果如图7E所示，RB-vTALE-AtKYP转基因拟南芥植物中CaLCuV的共同区(CommonRegion，CR)，互补链编码复制酶蛋白的基因AC1和互补链编码参与病毒复制或转录调控蛋白的基因AC4的表达水平至少降到载体对照植物中的1/10，说明RB-vTALE-AtKYP转基因拟南芥植物中RB-vTALE-AtKYP分子通过对组蛋白H3第9位赖氨酸进行甲基化修饰从而提高了DNA甲基化修饰的水平，最终抑制了病毒相关基因的表达，使得植物获得抗病毒的能力。
结论：CaLCuV特异的靶点RB-vTALE融合表达拟南芥组蛋白甲基转移酶KYP的RB-vTALE-AtKYP分子在双子叶植物拟南芥中稳定表达后对入侵的CaLCuV基因组中的RB-vTALE靶点所在核小体及其毗邻的核小体组蛋白进行甲基化修饰，使得核小体DNA受到进一步的甲基化修饰，从而抑制CaLCuV基因组的正常复制和转录过程。
表5.本发明中用于测定组蛋白和DNA甲基化程度的引物序列
实施例8.T4代RB-vTALE-AtKYP转基因拟南芥植物对甘蓝曲叶病毒(CaLCuV)的抗性效果、植物内源转座子Mu1和休眠相关基因的表达谱的影响。
待T4代转基因植物种子成熟后，挑选载体对照RB-vTALE-MCS和抗性比较好的RB-vTALE-AtKYP转基因株系的种子萌发T4代植株，按照实施例2中所述的注射接种本氏烟草的方法将CaLCuV侵染性克隆接种到4周左右的转基因拟南芥轮作叶上，观察发病情况。
接种CalCuV 21天后，T4代RB-vTALE-AtKYP转基因株系#1-12表现出高抗表型，植株生长健康，叶片无明显的斑驳或褪绿的症状(图8B)，而T4代载体对照RB-TALE-MCS#1转基因株系表现出易感表型，花叶及明显的褪绿等症状(图8A)。
利用病情指数这一指标按发病的早、中和晚期三个时间点统计的RB-vTALE-AtKYP转基因株系对CaLCuV的抗性效果如图8C所示，RB-vTALE-AtKYP转基因株系(#1-4和#1-12)在发病的早、中和晚期三个时间点均表现出高抗CaLCuV侵染的表型。
为了进一步验证观察到的结果，我们按照实施例2中所述的病毒滴度测定方法以拟南芥Tubulin2为内参基因(引物见表2)对接种CalCuV的转基因拟南芥进行病毒滴度的测定。
如图8D所示，病毒滴度分析的结果表明RB-vTALE-AtKYP转基因植物株系(#1-4和#1-12)中CaLCuV的病毒滴度明显低于载体对照植物，是其1/16，说明RB-vTALE-AtKYP转基因植物株系(#1-4和#1-12)对CaLCuV的侵染具有抗性效果，这一结果与观察到的发病症状及病情指数调查的结果相一致。
在拟南芥基因组内部含有丰富的转座子，为了检测RB-vTALE-AtKYP转基因植物株系中AtKYP的组蛋白甲基化修饰及由其引发的DNA甲基化修饰是否会脱靶作用在转座子上，我们按照实施例6中检测AtKYP基因表达水平的方法，检测拟南芥基因组中的转座子Mu1在T4代RB-vTALE-AtKYP转基因植物株系(#1-4和#1-12)中的表达水平(引物序列见表4)。如果RB-vTALE-AtKYP转基因植物株系中RB-vTALE-AtKYP分子脱靶即没有结合在RB-TALE靶点位置而是随机结合在Mu1的启动子区，那么AtKYP就会对Mu1启动子区进行组蛋白甲基化修饰，进而引起DNA甲基化修饰，最终抑制Mu1的转录，但是实际上拟南芥基因组中的转座子Mu1在RB-vTALE-AtKYP转基因植物株系(#1-4和#1-12)中和载体对照植物中的表达水平并无明显差异(图8E)，说明RB-vTALE-AtKYP转基因植物中RB-vTALE-AtKYP分子并未脱靶作用于拟南芥基因组的转座子Mu1上。
据文献(Jian Zheng et al.，2012，New phytologist)报道，AtKYP与种子休眠呈负相关，为了进一步验证RB-vTALE-AtKYP转基因植物株系中RB-vTALE-AtKYP分子是否存在脱靶的可能，我们检测T4代RB-vTALE-AtKYP转基因植物株系#1-12中与种子休眠相关基因的表达水平(引物序列见表4)。如图8F所示，种子休眠相关基因(AtABI3，AtDOG1和AtNCED6)在RB-vTALE-AtKYP转基因植物株系#1-12中和载体对照植物中的表达水平并无明显差异，说明RB-vTALE-AtKYP转基因植物中RB-vTALE-AtKYP分子并未脱靶作用于拟南芥基因组种子休眠相关基因。
结论：CaLCuV特异的靶点RB-vTALE融合表达拟南芥组蛋白甲基转移酶基因AtKYP的RB-vTALE-AtKYP分子在双子叶植物拟南芥中稳定表达后对CaLCuV的高抗表型能够稳定遗传给第四代，且RB-vTALE-AtKYP分子并未脱靶作用于拟南芥的转座子Mu1和种子休眠相关的基因上。
应该理解，尽管参考其示例性的实施方案，已经对本发明进行具体地显示和描述，但是本领域的普通技术人员应该理解，在不背离由后附的权利要求所定义的本发明的精神和范围的条件下，可以在其中进行各种形式和细节的变化，可以进行各种实施方案的任意组合。
序列表
<110> 中国科学院微生物研究所
<120> 一种提高植物对双生病毒抗性的方法
<130> IB168044
<160> 30
<170> PatentIn version 3.1
<210> 1
<211> 18
<212> DNA
<213> IR-vTALE靶点序列
<400> 1
taatattacc ggatggcc 18
<210> 2
<211> 18
<212> DNA
<213> RB-vTALE靶点序列
<400> 2
tatatattgg acaccaag 18
<210> 3
<211> 1875
<212> DNA
<213> 来自拟南芥的AtKYP核苷酸序列
<400> 3
atggctggaa aaaggaaacg agctaatgct cctgaccaaa cagagcgaag atcgagtgtt 60
cgggttcaga aagtgagaca gaaagcgtta gatgagaagg cgcgtttagt acaggagagg 120
gttaagctcc tcagtgacag aaagagtgaa atttgtgtcg atgacactga gttacatgag 180
aaagaagagg aaaatgtcga tgggagccct aaacgaagaa gccctccaaa gctaaccgca 240
atgcagaaag gaaagcagaa attgagtgtt tctctgaatg gtaaggacgt gaacttggaa 300
cctcatctca aagtgacaaa gtgtctgagg ttatttaaca agcaatatct cctctgtgtc 360
caggctaagt tgagcaggcc tgatttgaag ggtgtaactg agatgataaa agctaaggcg 420
atattgtacc caagaaaaat aatcggtgac cttccaggta tagacgttgg acaccgtttt 480
ttttcaagag ctgaaatgtg tgctgtagga ttccataacc attggctaaa tggcattgat 540
tatatgtcaa tggaatacga aaaagagtat agtaactaca aattaccgct tgctgtttct 600
attgttatgt cgggccagta cgaggatgat ctagacaatg cagatacagt gacttacact 660
ggacagggag ggcataactt aactggtaat aaacgtcaga taaaggatca acttttagaa 720
cgagggaatt tggcgctaaa gcactgctgc gaatataatg tgcctgtcag agtaactcgt 780
ggtcacaatt gcaaaagtag ctataccaaa cgagtataca cttatgatgg actgtacaag 840
gttgaaaagt tctgggcaca aaagggcgtt tcaggattta cagtgtataa gtaccgactg 900
aaacgattgg aggggcaacc agaactaact actgatcagg tcaactttgt tgctggacgc 960
ataccaacga gtacttcaga aattgagggt ttggtatgtg aggacatctc cggagggcta 1020
gaatttaagg gtatccccgc cactaatcgt gttgatgatt caccagtttc accaacatct 1080
ggtttcacat acatcaaatc tttgattatt gagcctaatg tcataattcc aaagagttca 1140
actgggtgta actgccgagg cagctgcact gactcaaaga aatgtgcatg tgctaagctt 1200
aatgggggta actttccata tgttgacctt aatgatggca gattaattga gtctcgagat 1260
gttgtatttg aatgtggtcc tcactgtggg tgtgggccaa aatgtgtcaa ccgaacttct 1320
cagaagcgtc taagattcaa tcttgaggtt ttccgctctg caaagaaggg ttgggcagtt 1380
agatcatggg agtacatacc agctggttca ccagtatgtg agtacatagg agttgtcagg 1440
agaactgctg atgtggatac tatctctgac aatgaataca tatttgagat tgactgccaa 1500
cagacaatgc aaggtcttgg tggaagacag agaagactaa gagatgttgc tgtaccaatg 1560
aataatggag tcagtcagag cagtgaagat gagaatgcgc cagagttctg cattgatgct 1620
ggttcaacag gaaactttgc taggtttata aatcacagtt gtgaaccaaa cctatttgtt 1680
cagtgcgtcc tgagttctca ccaggatata aggcttgccc gtgtggttct tttcgcagct 1740
gacaacattt ccccaatgca ggagctcact tacgactatg gatatgcgct tgatagcgtt 1800
catggaccgg atgggaaggt gaagcagctc gcttgctact gtggagcgct aaattgtagg 1860
aaacgccttt actaa 1875
<210> 4
<211> 1410
<212> DNA
<213> 来源于拟南芥的AtHDAC19核苷酸序列
<400> 4
atggatactg gcggcaattc gctggcgtcc ggacctgatg gtgtgaagag gaaagtttgt 60
tatttctatg accctgaggt cggcaattac tactatggcc aaggtcatcc catgaagccc 120
catcgcatcc gcatgaccca tgccctcctc gctcactacg gtctccttca gcatatgcag 180
gttctcaagc ccttccctgc ccgcgaccgt gatctctgcc gcttccacgc cgacgactat 240
gtctcttttc tccgcagcat tacccctgaa acccagcaag atcagattcg ccaacttaag 300
cgcttcaatg ttggtgaaga ctgtcccgtc tttgacggcc tttattcctt ttgccagacc 360
tatgctggag gatctgttgg tggctctgtc aagcttaacc acggcctctg cgatattgcc 420
atcaactggg ctggtggtct ccatcacgct aagaagtgcg aggcctctgg cttctgttac 480
gtcaatgata tcgtcttagc tatcctagag ctccttaagc agcatgagcg tgttctttat 540
gtcgatattg atatccacca cggggatgga gtggaggagg cattttatgc tactgacagg 600
gttatgactg tctcgtttca taaatttggt gattactttc ccggtacagg tcacattcag 660
gatataggtt atggtagcgg aaagtactat tctctcaatg taccactgga tgatggaatc 720
gatgatgaga gctatcatct gttattcaag cccatcatgg ggaaagttat ggaaattttc 780
cgaccagggg ctgtggtatt gcaatgtggt gctgattcat tgtctggtga taggttgggg 840
tgctttaatc tttcaatcaa aggtcatgct gagtgcgtca aatttatgag atcgttcaat 900
gttcccctac tgctcttggg tggtggtggt tacactatcc gcaatgttgc ccgttgctgg 960
tgctacgaga ctggagttgc acttggagtt gaagttgaag acaagatgcc ggagcatgaa 1020
tattatgaat actttggtcc agactataca cttcacgttg ctccaagtaa catggaaaat 1080
aagaattctc gtcagatgct tgaagagatt cgcaatgacc ttctccacaa tctctctaag 1140
cttcagcatg ctccaagtgt accatttcag gaaagaccac ctgatacaga gactcccgag 1200
gttgatgaag accaagaaga tggggataaa agatgggatc cggattcaga catggatgtt 1260
gatgatgacc gtaaacctat accaagcaga gtaaaaagag aagctgttga accagataca 1320
aaggacaagg atggactgaa aggaattatg gagcgtggaa aaggttgtga ggtggaggtg 1380
gatgagagtg gaagcactaa ggtaaaatga 1410
<210> 5
<211> 4605
<212> DNA
<213> 来源于拟南芥的AtMET1核苷酸序列
<400> 5
atggtggaaa atggggctaa agctgcgaag cgaaagaaga gaccacttcc agagattcaa 60
gaggtagaag atgtacctag gacgaggaga ccaaggcgtg ctgcagcgtg taccagtttc 120
aaggagaaat ctattcgagt ctgtgagaaa tctgctacta ttgaagtaaa gaaacagcag 180
attgtggagg aagagtttct cgcgttacgg ttaacggctc tggaaactga tgttgaagat 240
cgtccaacca ggagactgaa tgattttgtt ttgtttgatt cagatggagt tccacaacct 300
ctggagatgt tggagattca tgacatattc gtttcaggtg ctatcttacc ttcagatgtg 360
tgtactgata aggagaaaga gaagggtgtg aggtgtacat cgtttggacg ggttgagcat 420
tggagtatct ctggttatga agatggttcc cctgttattt ggatctcaac ggaattggcg 480
gattatgatt gtcgtaaacc tgctgctagc tacaggaagg tttatgatta cttctatgag 540
aaagctcgtg cttcagtggc tgtgtataag aaattgtcca agtcatctgg tggggatcct 600
gatataggtc ttgaggagtt acttgcggcg gttgtcagat caatgagcag tggaagcaag 660
tacttttcta gtggtgcggc aatcatcgat tttgttatat cccagggaga ttttatatat 720
aaccaactcg ctggtttgga tgagacagcc aagaaacatg aatcaagcta tgttgagatt 780
cctgttcttg tagctctcag agagaagagt agtaagattg acaagcctct gcagagggaa 840
agaaacccat ctaatggtgt gaggattaaa gaagtttctc aagttgcgga gagcgaggcc 900
ttgacatctg atcaactggt tgatggtact gatgatgaca gaagatatgc tatactctta 960
caagacgaag agaataggaa atctatgcaa cagcccagaa aaaacagcag ctcaggttct 1020
gcttcaaata tgttctacat taagataaat gaagatgaga ttgccaatga ttatcctctc 1080
ccatcgtact ataagacctc cgaagaagaa acagatgaac ttatacttta tgatgcttcc 1140
tatgaggttc aatctgaaca cctgcctcac aggatgcttc acaactgggc tctttataac 1200
tctgatttac gattcatatc actggaactt ctaccgatga aacaatgtga tgatattgat 1260
gtcaacattt ttgggtcagg tgtggtgact gatgataatg gaagttggat ttctttaaac 1320
gatcctgaca gcggttctca gtcacacgat cctgatggga tgtgcatatt cctcagtcaa 1380
attaaagaat ggatgattga gtttgggagc gatgatatta tctccatttc tatacgaaca 1440
gatgtggcct ggtaccgtct tgggaaacca tcaaaacttt atgccccttg gtggaaacct 1500
gttctgaaaa cagcaagggt tgggataagc attcttactt ttcttagggt ggaaagtagg 1560
gttgctaggc tttcatttgc agatgtcaca aaaagactgt ctgggttaca ggcgaatgat 1620
aaagcttaca tttcttctga ccccttggct gttgagagat atttggtcgt ccatgggcaa 1680
attattttac agctttttgc agtttatccg gacgacaatg tcaaaaggtg tccatttgtt 1740
gttggtcttg caagcaaatt ggaggatagg caccacacaa aatggatcat caagaagaag 1800
aaaatttcgc tgaaggaact gaatctgaat ccaagggcag gcatggcacc agtagcatcg 1860
aagaggaaag ctatgcaagc aacaacaact cgcctggtca acagaatttg gggagagttt 1920
tactccaatt actctccaga ggatccattg caggcgactg ctgcagaaaa tggggaggat 1980
gaggtggaag aggaaggcgg aaatggggag gaagaggttg aagaggaagg tgaaaatggt 2040
ctcacagagg acactgtacc agaacctgtt gaggttcaga agcctcatac tcctaagaaa 2100
atccgaggca gttctggaaa aagggaaata aaatgggatg gtgagagtct aggaaaaact 2160
tctgctggcg agcctctcta tcaacaagcc cttgttggag gggaaatggt ggctgtaggt 2220
ggcgctgtca ccttggaagt tgatgatcca gatgaaatgc cggccatcta ttttgtggag 2280
tacatgttcg aaagtacaga tcactgcaaa atgttacatg gtagattctt acaaagagga 2340
tctatgactg ttctggggaa tgctgctaac gagagggaac tattcctgac taatgaatgc 2400
atgactacac agctcaagga cattaaagga gtagccagtt ttgagattcg atcaaggcca 2460
tgggggcatc agtataggaa aaagaacatc actgcggata agcttgactg ggctagagca 2520
ttagaaagaa aagtaaaaga tttgccaaca gagtattact gcaaaagctt gtactcacct 2580
gagagagggg gattctttag tcttccacta agtgatattg gtcgcagttc tgggttctgc 2640
acttcatgta agataaggga ggatgaagag aagaggtcta caattaaact aaatgtttca 2700
aagacaggct ttttcatcaa tgggattgag tattctgttg aggattttgt ctatgtcaac 2760
cctgactcta ttggtgggtt gaaggagggt agtaaaactt cttttaagtc tgggcgaaac 2820
attgggttaa gagcgtatgt tgtttgccaa ttgctggaaa ttgttccaaa ggaatctaga 2880
aaggctgatt tgggttcctt tgatgttaaa gtgagaaggt tttataggcc tgaggatgtt 2940
tctgcagaga aggcctatgc ttcagacatc caagaattgt atttcagcca ggacacagtt 3000
gttctccctc caggtgctct agagggaaaa tgtgaagtaa gaaagaaaag tgatatgccc 3060
ttatcccgtg aatatccaat atcagaccat attttcttct gtgatctttt ctttgacacc 3120
tccaaaggtt ctctcaagca gctgcccgcc aatatgaagc caaagttctc tactattaag 3180
gacgacacac ttttaagaaa gaaaaaggga aagggagtag agagtgaaat tgagtctgag 3240
attgtcaagc ctgttgagcc acctaaagag attcgtctgg ctactctaga tatttttgct 3300
ggttgtggtg gcctgtctca tggactgaaa aaggcgggtg tatctgatgc aaagtgggcg 3360
attgagtatg aagagccagc tgggcaggct tttaaacaaa accatcctga gtcaacagtt 3420
tttgttgaca actgcaatgt gattcttagg gctataatgg agaaaggtgg agatcaagat 3480
gattgtgtct ctactacaga ggcaaatgaa ttagcagcta aactaactga ggagcagaag 3540
agtactctgc cactgcctgg tcaagtggac ttcatcaatg gtggacctcc atgtcaggga 3600
ttttctggta tgaacaggtt caaccaaagc tcttggagta aagttcagtg tgaaatgata 3660
ttagcattct tgtcctttgc tgactatttc cggccaaggt attttcttct ggagaacgtg 3720
aggacctttg tgtcattcaa taaagggcag acatttcagc ttactttggc ttcccttctc 3780
gaaatgggtt accaggtgag atttggaatc ctggaggccg gtgcatatgg agtatcccaa 3840
tctcgtaaac gagctttcat ttgggctgct gcaccagaag aagttctccc tgaatggcct 3900
gagccgatgc atgtctttgg tgttccaaag ttgaaaatct cactatctca aggtttacat 3960
tatgctgctg ttcgtagtac tgcacttggt gcccctttcc gtccaatcac cgtgagagac 4020
acaattggtg atcttccatc agtagaaaac ggagactcta ggacaaacaa agagtataaa 4080
gaggttgcag tctcgtggtt ccaaaaggag ataagaggaa acacgattgc tctcactgat 4140
catatctgca aggctatgaa tgagcttaac ctcattcgat gcaaattaat cccaactagg 4200
cctggggctg attggcatga cttgccaaag agaaaggtta cgttatctga tgggcgcgta 4260
gaagaaatga ttcctttttg tctcccaaac acagctgagc gccacaacgg ttggaaggga 4320
ctatatggga gattagattg gcaaggaaac tttccgactt ccgtcacgga tcctcagccc 4380
atgggtaagg ttggaatgtg ctttcatcct gaacagcaca gaatccttac agtccgtgaa 4440
tgcgcccgat ctcaggggtt tccggatagc tacgagtttg cagggaacat aaatcacaag 4500
cacaggcaga ttgggaatgc agtccctcca ccattggcat ttgctctagg tcgtaagctc 4560
aaagaagccc tacatctcaa gaagtctcct caacaccaac cctag 4605
<210> 6
<211> 36
<212> DNA
<213> 来源于拟南芥的AtSRDX核苷酸序列
<400> 6
cttgatctgg atctggaact ccgtttgggt ttcgct 36
<210> 7
<211> 1956
<212> DNA
<213> 来源于拟南芥的AtSUVH2核苷酸序列
<400> 7
atgagtacat tgttaccatt tcctgacctc aacctcatgc cggattctca atcctccacc 60
gccggaacca cagccggcga cactgtagtc accggaaagt tagaagtgaa atcggaacca 120
attgaggaat ggcaaactcc accatcgtct acctccgatc aatcagccaa taccgatctc 180
atcgccgagt ttattcgtat ctcagagctc ttccgctcgg cattcaagcc actgcaggtc 240
aaaggattag acggagtctc cgtatacgga ttggattctg gtgccatcgt tgctgtaccg 300
gagaaagaaa accgggaatt gattgaaccg cctccaggat ttaaggataa tcgagtctcc 360
accgtcgttg tctcgccgaa attcgagaga ccgagagagc tagcgagaat tgcaattcta 420
ggtcatgaac aacggaagga actccgacaa gtcatgaaac gaactaggat gacttatgag 480
tctcttcgga ttcatctaat ggcggagagt atgaagaatc atgtacttgg tcaagggcgt 540
agacggagga gcgatatggc ggctgcgtat ataatgaggg atcggggact gtggttgaac 600
tatgataagc acatagttgg tccagtcaca ggggttgaag taggggatat attcttctac 660
cgtatggaat tgtgtgtgct gggcttacac ggccagacac aagccgggat tgattgttta 720
acggctgaac ggagcgccac tggagagcct atagctacca gcattgttgt ctcaggtggt 780
tatgaggatg atgaagatac aggagatgtt ttggtttata ctggtcacgg tggtcaggat 840
catcaacaca agcagtgtga taaccagagg ctagtaggtg ggaatctggg aatggagaga 900
agtatgcact acggtatcga ggtacgtgtg attagaggta ttaagtatga gaacagtata 960
agctcgaagg tgtatgtata tgatggtttg tataagatag tagattggtg gtttgctgtg 1020
gggaagtctg gttttggggt ttttaaattt aggttagtaa gaattgaagg gcaaccgatg 1080
atgggcagtg cggtaatgag gtttgcacaa actcttagaa ataagccatc gatggttagg 1140
cctactggtt atgttagctt tgacctttct aacaagaagg agaatgtacc cgtgtttcta 1200
tataatgatg tagatggtga tcaagaaccg agacattatg agtacattgc aaaagctgtc 1260
tttcctcctg gaatttttgg tcaggggggg atcagcagga ctggctgcga gtgtaagctc 1320
tcttgtactg atgattgtct ctgtgcaagg aagaacggtg gtgagtttgc atatgatgat 1380
aacgggcatc tattgaaagg aaagcatgtg gtatttgaat gtggggaatt ctgcacttgt 1440
ggtccgagct gtaagagccg tgtgacacag aagggattga ggaacaggct agaggttttc 1500
agatcgaagg aaaccggttg gggtgtcagg acacttgatc taattgaagc tggtgctttc 1560
atatgcgaat atgcaggtgt agttgtcacg agacttcaag ccgagattct gtcaatgaat 1620
ggggatgtta tggtctatcc tggtcggttc acagaccaat ggcgtaactg gggtgattta 1680
tctcaagtat acccagattt tgttaggccg aattatccat ctctccctcc actggatttc 1740
tcaatggatg tgtcaaggat gaggaacgtg gcttgctaca ttagtcacag caaagaacca 1800
aatgtgatgg tgcaatttgt tttgcatgac cacaatcacc tgatgttccc ccgtgtgatg 1860
ctttttgcgc tggagaatat ctcaccgttg gccgagctaa gcctggatta cggcttggca 1920
gatgaagtga acggcaagct cgccatctgc aactag 1956
<210> 8
<211> 624
<212> PRT
<213> 来源于拟南芥的AtKYP氨基酸序列
<400> 8
Met Ala Gly Lys Arg Lys Arg Ala Asn Ala Pro Asp Gln Thr Glu Arg
1 5 10 15
Arg Ser Ser Val Arg Val Gln Lys Val Arg Gln Lys Ala Leu Asp Glu
20 25 30
Lys Ala Arg Leu Val Gln Glu Arg Val Lys Leu Leu Ser Asp Arg Lys
35 40 45
Ser Glu Ile Cys Val Asp Asp Thr Glu Leu His Glu Lys Glu Glu Glu
50 55 60
Asn Val Asp Gly Ser Pro Lys Arg Arg Ser Pro Pro Lys Leu Thr Ala
65 70 75 80
Met Gln Lys Gly Lys Gln Lys Leu Ser Val Ser Leu Asn Gly Lys Asp
85 90 95
Val Asn Leu Glu Pro His Leu Lys Val Thr Lys Cys Leu Arg Leu Phe
100 105 110
Asn Lys Gln Tyr Leu Leu Cys Val Gln Ala Lys Leu Ser Arg Pro Asp
115 120 125
Leu Lys Gly Val Thr Glu Met Ile Lys Ala Lys Ala Ile Leu Tyr Pro
130 135 140
Arg Lys Ile Ile Gly Asp Leu Pro Gly Ile Asp Val Gly His Arg Phe
145 150 155 160
Phe Ser Arg Ala Glu Met Cys Ala Val Gly Phe His Asn His Trp Leu
165 170 175
Asn Gly Ile Asp Tyr Met Ser Met Glu Tyr Glu Lys Glu Tyr Ser Asn
180 185 190
Tyr Lys Leu Pro Leu Ala Val Ser Ile Val Met Ser Gly Gln Tyr Glu
195 200 205
Asp Asp Leu Asp Asn Ala Asp Thr Val Thr Tyr Thr Gly Gln Gly Gly
210 215 220
His Asn Leu Thr Gly Asn Lys Arg Gln Ile Lys Asp Gln Leu Leu Glu
225 230 235 240
Arg Gly Asn Leu Ala Leu Lys His Cys Cys Glu Tyr Asn Val Pro Val
245 250 255
Arg Val Thr Arg Gly His Asn Cys Lys Ser Ser Tyr Thr Lys Arg Val
260 265 270
Tyr Thr Tyr Asp Gly Leu Tyr Lys Val Glu Lys Phe Trp Ala Gln Lys
275 280 285
Gly Val Ser Gly Phe Thr Val Tyr Lys Tyr Arg Leu Lys Arg Leu Glu
290 295 300
Gly Gln Pro Glu Leu Thr Thr Asp Gln Val Asn Phe Val Ala Gly Arg
305 310 315 320
Ile Pro Thr Ser Thr Ser Glu Ile Glu Gly Leu Val Cys Glu Asp Ile
325 330 335
Ser Gly Gly Leu Glu Phe Lys Gly Ile Pro Ala Thr Asn Arg Val Asp
340 345 350
Asp Ser Pro Val Ser Pro Thr Ser Gly Phe Thr Tyr Ile Lys Ser Leu
355 360 365
Ile Ile Glu Pro Asn Val Ile Ile Pro Lys Ser Ser Thr Gly Cys Asn
370 375 380
Cys Arg Gly Ser Cys Thr Asp Ser Lys Lys Cys Ala Cys Ala Lys Leu
385 390 395 400
Asn Gly Gly Asn Phe Pro Tyr Val Asp Leu Asn Asp Gly Arg Leu Ile
405 410 415
Glu Ser Arg Asp Val Val Phe Glu Cys Gly Pro His Cys Gly Cys Gly
420 425 430
Pro Lys Cys Val Asn Arg Thr Ser Gln Lys Arg Leu Arg Phe Asn Leu
435 440 445
Glu Val Phe Arg Ser Ala Lys Lys Gly Trp Ala Val Arg Ser Trp Glu
450 455 460
Tyr Ile Pro Ala Gly Ser Pro Val Cys Glu Tyr Ile Gly Val Val Arg
465 470 475 480
Arg Thr Ala Asp Val Asp Thr Ile Ser Asp Asn Glu Tyr Ile Phe Glu
485 490 495
Ile Asp Cys Gln Gln Thr Met Gln Gly Leu Gly Gly Arg Gln Arg Arg
500 505 510
Leu Arg Asp Val Ala Val Pro Met Asn Asn Gly Val Ser Gln Ser Ser
515 520 525
Glu Asp Glu Asn Ala Pro Glu Phe Cys Ile Asp Ala Gly Ser Thr Gly
530 535 540
Asn Phe Ala Arg Phe Ile Asn His Ser Cys Glu Pro Asn Leu Phe Val
545 550 555 560
Gln Cys Val Leu Ser Ser His Gln Asp Ile Arg Leu Ala Arg Val Val
565 570 575
Leu Phe Ala Ala Asp Asn Ile Ser Pro Met Gln Glu Leu Thr Tyr Asp
580 585 590
Tyr Gly Tyr Ala Leu Asp Ser Val His Gly Pro Asp Gly Lys Val Lys
595 600 605
Gln Leu Ala Cys Tyr Cys Gly Ala Leu Asn Cys Arg Lys Arg Leu Tyr
610 615 620
<210> 9
<211> 469
<212> PRT
<213> 来源于拟南芥的AtHDAC19氨基酸序列
<400> 9
Met Asp Thr Gly Gly Asn Ser Leu Ala Ser Gly Pro Asp Gly Val Lys
1 5 10 15
Arg Lys Val Cys Tyr Phe Tyr Asp Pro Glu Val Gly Asn Tyr Tyr Tyr
20 25 30
Gly Gln Gly His Pro Met Lys Pro His Arg Ile Arg Met Thr His Ala
35 40 45
Leu Leu Ala His Tyr Gly Leu Leu Gln His Met Gln Val Leu Lys Pro
50 55 60
Phe Pro Ala Arg Asp Arg Asp Leu Cys Arg Phe His Ala Asp Asp Tyr
65 70 75 80
Val Ser Phe Leu Arg Ser Ile Thr Pro Glu Thr Gln Gln Asp Gln Ile
85 90 95
Arg Gln Leu Lys Arg Phe Asn Val Gly Glu Asp Cys Pro Val Phe Asp
100 105 110
Gly Leu Tyr Ser Phe Cys Gln Thr Tyr Ala Gly Gly Ser Val Gly Gly
115 120 125
Ser Val Lys Leu Asn His Gly Leu Cys Asp Ile Ala Ile Asn Trp Ala
130 135 140
Gly Gly Leu His His Ala Lys Lys Cys Glu Ala Ser Gly Phe Cys Tyr
145 150 155 160
Val Asn Asp Ile Val Leu Ala Ile Leu Glu Leu Leu Lys Gln His Glu
165 170 175
Arg Val Leu Tyr Val Asp Ile Asp Ile His His Gly Asp Gly Val Glu
180 185 190
Glu Ala Phe Tyr Ala Thr Asp Arg Val Met Thr Val Ser Phe His Lys
195 200 205
Phe Gly Asp Tyr Phe Pro Gly Thr Gly His Ile Gln Asp Ile Gly Tyr
210 215 220
Gly Ser Gly Lys Tyr Tyr Ser Leu Asn Val Pro Leu Asp Asp Gly Ile
225 230 235 240
Asp Asp Glu Ser Tyr His Leu Leu Phe Lys Pro Ile Met Gly Lys Val
245 250 255
Met Glu Ile Phe Arg Pro Gly Ala Val Val Leu Gln Cys Gly Ala Asp
260 265 270
Ser Leu Ser Gly Asp Arg Leu Gly Cys Phe Asn Leu Ser Ile Lys Gly
275 280 285
His Ala Glu Cys Val Lys Phe Met Arg Ser Phe Asn Val Pro Leu Leu
290 295 300
Leu Leu Gly Gly Gly Gly Tyr Thr Ile Arg Asn Val Ala Arg Cys Trp
305 310 315 320
Cys Tyr Glu Thr Gly Val Ala Leu Gly Val Glu Val Glu Asp Lys Met
325 330 335
Pro Glu His Glu Tyr Tyr Glu Tyr Phe Gly Pro Asp Tyr Thr Leu His
340 345 350
Val Ala Pro Ser Asn Met Glu Asn Lys Asn Ser Arg Gln Met Leu Glu
355 360 365
Glu Ile Arg Asn Asp Leu Leu His Asn Leu Ser Lys Leu Gln His Ala
370 375 380
Pro Ser Val Pro Phe Gln Glu Arg Pro Pro Asp Thr Glu Thr Pro Glu
385 390 395 400
Val Asp Glu Asp Gln Glu Asp Gly Asp Lys Arg Trp Asp Pro Asp Ser
405 410 415
Asp Met Asp Val Asp Asp Asp Arg Lys Pro Ile Pro Ser Arg Val Lys
420 425 430
Arg Glu Ala Val Glu Pro Asp Thr Lys Asp Lys Asp Gly Leu Lys Gly
435 440 445
Ile Met Glu Arg Gly Lys Gly Cys Glu Val Glu Val Asp Glu Ser Gly
450 455 460
Ser Thr Lys Val Lys
465
<210> 10
<211> 1534
<212> PRT
<213> 来源于拟南芥的AtMET1氨基酸序列
<400> 10
Met Val Glu Asn Gly Ala Lys Ala Ala Lys Arg Lys Lys Arg Pro Leu
1 5 10 15
Pro Glu Ile Gln Glu Val Glu Asp Val Pro Arg Thr Arg Arg Pro Arg
20 25 30
Arg Ala Ala Ala Cys Thr Ser Phe Lys Glu Lys Ser Ile Arg Val Cys
35 40 45
Glu Lys Ser Ala Thr Ile Glu Val Lys Lys Gln Gln Ile Val Glu Glu
50 55 60
Glu Phe Leu Ala Leu Arg Leu Thr Ala Leu Glu Thr Asp Val Glu Asp
65 70 75 80
Arg Pro Thr Arg Arg Leu Asn Asp Phe Val Leu Phe Asp Ser Asp Gly
85 90 95
Val Pro Gln Pro Leu Glu Met Leu Glu Ile His Asp Ile Phe Val Ser
100 105 110
Gly Ala Ile Leu Pro Ser Asp Val Cys Thr Asp Lys Glu Lys Glu Lys
115 120 125
Gly Val Arg Cys Thr Ser Phe Gly Arg Val Glu His Trp Ser Ile Ser
130 135 140
Gly Tyr Glu Asp Gly Ser Pro Val Ile Trp Ile Ser Thr Glu Leu Ala
145 150 155 160
Asp Tyr Asp Cys Arg Lys Pro Ala Ala Ser Tyr Arg Lys Val Tyr Asp
165 170 175
Tyr Phe Tyr Glu Lys Ala Arg Ala Ser Val Ala Val Tyr Lys Lys Leu
180 185 190
Ser Lys Ser Ser Gly Gly Asp Pro Asp Ile Gly Leu Glu Glu Leu Leu
195 200 205
Ala Ala Val Val Arg Ser Met Ser Ser Gly Ser Lys Tyr Phe Ser Ser
210 215 220
Gly Ala Ala Ile Ile Asp Phe Val Ile Ser Gln Gly Asp Phe Ile Tyr
225 230 235 240
Asn Gln Leu Ala Gly Leu Asp Glu Thr Ala Lys Lys His Glu Ser Ser
245 250 255
Tyr Val Glu Ile Pro Val Leu Val Ala Leu Arg Glu Lys Ser Ser Lys
260 265 270
Ile Asp Lys Pro Leu Gln Arg Glu Arg Asn Pro Ser Asn Gly Val Arg
275 280 285
Ile Lys Glu Val Ser Gln Val Ala Glu Ser Glu Ala Leu Thr Ser Asp
290 295 300
Gln Leu Val Asp Gly Thr Asp Asp Asp Arg Arg Tyr Ala Ile Leu Leu
305 310 315 320
Gln Asp Glu Glu Asn Arg Lys Ser Met Gln Gln Pro Arg Lys Asn Ser
325 330 335
Ser Ser Gly Ser Ala Ser Asn Met Phe Tyr Ile Lys Ile Asn Glu Asp
340 345 350
Glu Ile Ala Asn Asp Tyr Pro Leu Pro Ser Tyr Tyr Lys Thr Ser Glu
355 360 365
Glu Glu Thr Asp Glu Leu Ile Leu Tyr Asp Ala Ser Tyr Glu Val Gln
370 375 380
Ser Glu His Leu Pro His Arg Met Leu His Asn Trp Ala Leu Tyr Asn
385 390 395 400
Ser Asp Leu Arg Phe Ile Ser Leu Glu Leu Leu Pro Met Lys Gln Cys
405 410 415
Asp Asp Ile Asp Val Asn Ile Phe Gly Ser Gly Val Val Thr Asp Asp
420 425 430
Asn Gly Ser Trp Ile Ser Leu Asn Asp Pro Asp Ser Gly Ser Gln Ser
435 440 445
His Asp Pro Asp Gly Met Cys Ile Phe Leu Ser Gln Ile Lys Glu Trp
450 455 460
Met Ile Glu Phe Gly Ser Asp Asp Ile Ile Ser Ile Ser Ile Arg Thr
465 470 475 480
Asp Val Ala Trp Tyr Arg Leu Gly Lys Pro Ser Lys Leu Tyr Ala Pro
485 490 495
Trp Trp Lys Pro Val Leu Lys Thr Ala Arg Val Gly Ile Ser Ile Leu
500 505 510
Thr Phe Leu Arg Val Glu Ser Arg Val Ala Arg Leu Ser Phe Ala Asp
515 520 525
Val Thr Lys Arg Leu Ser Gly Leu Gln Ala Asn Asp Lys Ala Tyr Ile
530 535 540
Ser Ser Asp Pro Leu Ala Val Glu Arg Tyr Leu Val Val His Gly Gln
545 550 555 560
Ile Ile Leu Gln Leu Phe Ala Val Tyr Pro Asp Asp Asn Val Lys Arg
565 570 575
Cys Pro Phe Val Val Gly Leu Ala Ser Lys Leu Glu Asp Arg His His
580 585 590
Thr Lys Trp Ile Ile Lys Lys Lys Lys Ile Ser Leu Lys Glu Leu Asn
595 600 605
Leu Asn Pro Arg Ala Gly Met Ala Pro Val Ala Ser Lys Arg Lys Ala
610 615 620
Met Gln Ala Thr Thr Thr Arg Leu Val Asn Arg Ile Trp Gly Glu Phe
625 630 635 640
Tyr Ser Asn Tyr Ser Pro Glu Asp Pro Leu Gln Ala Thr Ala Ala Glu
645 650 655
Asn Gly Glu Asp Glu Val Glu Glu Glu Gly Gly Asn Gly Glu Glu Glu
660 665 670
Val Glu Glu Glu Gly Glu Asn Gly Leu Thr Glu Asp Thr Val Pro Glu
675 680 685
Pro Val Glu Val Gln Lys Pro His Thr Pro Lys Lys Ile Arg Gly Ser
690 695 700
Ser Gly Lys Arg Glu Ile Lys Trp Asp Gly Glu Ser Leu Gly Lys Thr
705 710 715 720
Ser Ala Gly Glu Pro Leu Tyr Gln Gln Ala Leu Val Gly Gly Glu Met
725 730 735
Val Ala Val Gly Gly Ala Val Thr Leu Glu Val Asp Asp Pro Asp Glu
740 745 750
Met Pro Ala Ile Tyr Phe Val Glu Tyr Met Phe Glu Ser Thr Asp His
755 760 765
Cys Lys Met Leu His Gly Arg Phe Leu Gln Arg Gly Ser Met Thr Val
770 775 780
Leu Gly Asn Ala Ala Asn Glu Arg Glu Leu Phe Leu Thr Asn Glu Cys
785 790 795 800
Met Thr Thr Gln Leu Lys Asp Ile Lys Gly Val Ala Ser Phe Glu Ile
805 810 815
Arg Ser Arg Pro Trp Gly His Gln Tyr Arg Lys Lys Asn Ile Thr Ala
820 825 830
Asp Lys Leu Asp Trp Ala Arg Ala Leu Glu Arg Lys Val Lys Asp Leu
835 840 845
Pro Thr Glu Tyr Tyr Cys Lys Ser Leu Tyr Ser Pro Glu Arg Gly Gly
850 855 860
Phe Phe Ser Leu Pro Leu Ser Asp Ile Gly Arg Ser Ser Gly Phe Cys
865 870 875 880
Thr Ser Cys Lys Ile Arg Glu Asp Glu Glu Lys Arg Ser Thr Ile Lys
885 890 895
Leu Asn Val Ser Lys Thr Gly Phe Phe Ile Asn Gly Ile Glu Tyr Ser
900 905 910
Val Glu Asp Phe Val Tyr Val Asn Pro Asp Ser Ile Gly Gly Leu Lys
915 920 925
Glu Gly Ser Lys Thr Ser Phe Lys Ser Gly Arg Asn Ile Gly Leu Arg
930 935 940
Ala Tyr Val Val Cys Gln Leu Leu Glu Ile Val Pro Lys Glu Ser Arg
945 950 955 960
Lys Ala Asp Leu Gly Ser Phe Asp Val Lys Val Arg Arg Phe Tyr Arg
965 970 975
Pro Glu Asp Val Ser Ala Glu Lys Ala Tyr Ala Ser Asp Ile Gln Glu
980 985 990
Leu Tyr Phe Ser Gln Asp Thr Val Val Leu Pro Pro Gly Ala Leu Glu
995 1000 1005
Gly Lys Cys Glu Val Arg Lys Lys Ser Asp Met Pro Leu Ser Arg
1010 1015 1020
Glu Tyr Pro Ile Ser Asp His Ile Phe Phe Cys Asp Leu Phe Phe
1025 1030 1035
Asp Thr Ser Lys Gly Ser Leu Lys Gln Leu Pro Ala Asn Met Lys
1040 1045 1050
Pro Lys Phe Ser Thr Ile Lys Asp Asp Thr Leu Leu Arg Lys Lys
1055 1060 1065
Lys Gly Lys Gly Val Glu Ser Glu Ile Glu Ser Glu Ile Val Lys
1070 1075 1080
Pro Val Glu Pro Pro Lys Glu Ile Arg Leu Ala Thr Leu Asp Ile
1085 1090 1095
Phe Ala Gly Cys Gly Gly Leu Ser His Gly Leu Lys Lys Ala Gly
1100 1105 1110
Val Ser Asp Ala Lys Trp Ala Ile Glu Tyr Glu Glu Pro Ala Gly
1115 1120 1125
Gln Ala Phe Lys Gln Asn His Pro Glu Ser Thr Val Phe Val Asp
1130 1135 1140
Asn Cys Asn Val Ile Leu Arg Ala Ile Met Glu Lys Gly Gly Asp
1145 1150 1155
Gln Asp Asp Cys Val Ser Thr Thr Glu Ala Asn Glu Leu Ala Ala
1160 1165 1170
Lys Leu Thr Glu Glu Gln Lys Ser Thr Leu Pro Leu Pro Gly Gln
1175 1180 1185
Val Asp Phe Ile Asn Gly Gly Pro Pro Cys Gln Gly Phe Ser Gly
1190 1195 1200
Met Asn Arg Phe Asn Gln Ser Ser Trp Ser Lys Val Gln Cys Glu
1205 1210 1215
Met Ile Leu Ala Phe Leu Ser Phe Ala Asp Tyr Phe Arg Pro Arg
1220 1225 1230
Tyr Phe Leu Leu Glu Asn Val Arg Thr Phe Val Ser Phe Asn Lys
1235 1240 1245
Gly Gln Thr Phe Gln Leu Thr Leu Ala Ser Leu Leu Glu Met Gly
1250 1255 1260
Tyr Gln Val Arg Phe Gly Ile Leu Glu Ala Gly Ala Tyr Gly Val
1265 1270 1275
Ser Gln Ser Arg Lys Arg Ala Phe Ile Trp Ala Ala Ala Pro Glu
1280 1285 1290
Glu Val Leu Pro Glu Trp Pro Glu Pro Met His Val Phe Gly Val
1295 1300 1305
Pro Lys Leu Lys Ile Ser Leu Ser Gln Gly Leu His Tyr Ala Ala
1310 1315 1320
Val Arg Ser Thr Ala Leu Gly Ala Pro Phe Arg Pro Ile Thr Val
1325 1330 1335
Arg Asp Thr Ile Gly Asp Leu Pro Ser Val Glu Asn Gly Asp Ser
1340 1345 1350
Arg Thr Asn Lys Glu Tyr Lys Glu Val Ala Val Ser Trp Phe Gln
1355 1360 1365
Lys Glu Ile Arg Gly Asn Thr Ile Ala Leu Thr Asp His Ile Cys
1370 1375 1380
Lys Ala Met Asn Glu Leu Asn Leu Ile Arg Cys Lys Leu Ile Pro
1385 1390 1395
Thr Arg Pro Gly Ala Asp Trp His Asp Leu Pro Lys Arg Lys Val
1400 1405 1410
Thr Leu Ser Asp Gly Arg Val Glu Glu Met Ile Pro Phe Cys Leu
1415 1420 1425
Pro Asn Thr Ala Glu Arg His Asn Gly Trp Lys Gly Leu Tyr Gly
1430 1435 1440
Arg Leu Asp Trp Gln Gly Asn Phe Pro Thr Ser Val Thr Asp Pro
1445 1450 1455
Gln Pro Met Gly Lys Val Gly Met Cys Phe His Pro Glu Gln His
1460 1465 1470
Arg Ile Leu Thr Val Arg Glu Cys Ala Arg Ser Gln Gly Phe Pro
1475 1480 1485
Asp Ser Tyr Glu Phe Ala Gly Asn Ile Asn His Lys His Arg Gln
1490 1495 1500
Ile Gly Asn Ala Val Pro Pro Pro Leu Ala Phe Ala Leu Gly Arg
1505 1510 1515
Lys Leu Lys Glu Ala Leu His Leu Lys Lys Ser Pro Gln His Gln
1520 1525 1530
Pro
<210> 11
<211> 12
<212> PRT
<213> 来源于拟南芥的AtSRDX氨基酸序列
<400> 11
Leu Asp Leu Asp Leu Glu Leu Arg Leu Gly Phe Ala
1 5 10
<210> 12
<211> 651
<212> PRT
<213> 来源于拟南芥的 AtSUVH2氨基酸序列
<400> 12
Met Ser Thr Leu Leu Pro Phe Pro Asp Leu Asn Leu Met Pro Asp Ser
1 5 10 15
Gln Ser Ser Thr Ala Gly Thr Thr Ala Gly Asp Thr Val Val Thr Gly
20 25 30
Lys Leu Glu Val Lys Ser Glu Pro Ile Glu Glu Trp Gln Thr Pro Pro
35 40 45
Ser Ser Thr Ser Asp Gln Ser Ala Asn Thr Asp Leu Ile Ala Glu Phe
50 55 60
Ile Arg Ile Ser Glu Leu Phe Arg Ser Ala Phe Lys Pro Leu Gln Val
65 70 75 80
Lys Gly Leu Asp Gly Val Ser Val Tyr Gly Leu Asp Ser Gly Ala Ile
85 90 95
Val Ala Val Pro Glu Lys Glu Asn Arg Glu Leu Ile Glu Pro Pro Pro
100 105 110
Gly Phe Lys Asp Asn Arg Val Ser Thr Val Val Val Ser Pro Lys Phe
115 120 125
Glu Arg Pro Arg Glu Leu Ala Arg Ile Ala Ile Leu Gly His Glu Gln
130 135 140
Arg Lys Glu Leu Arg Gln Val Met Lys Arg Thr Arg Met Thr Tyr Glu
145 150 155 160
Ser Leu Arg Ile His Leu Met Ala Glu Ser Met Lys Asn His Val Leu
165 170 175
Gly Gln Gly Arg Arg Arg Arg Ser Asp Met Ala Ala Ala Tyr Ile Met
180 185 190
Arg Asp Arg Gly Leu Trp Leu Asn Tyr Asp Lys His Ile Val Gly Pro
195 200 205
Val Thr Gly Val Glu Val Gly Asp Ile Phe Phe Tyr Arg Met Glu Leu
210 215 220
Cys Val Leu Gly Leu His Gly Gln Thr Gln Ala Gly Ile Asp Cys Leu
225 230 235 240
Thr Ala Glu Arg Ser Ala Thr Gly Glu Pro Ile Ala Thr Ser Ile Val
245 250 255
Val Ser Gly Gly Tyr Glu Asp Asp Glu Asp Thr Gly Asp Val Leu Val
260 265 270
Tyr Thr Gly His Gly Gly Gln Asp His Gln His Lys Gln Cys Asp Asn
275 280 285
Gln Arg Leu Val Gly Gly Asn Leu Gly Met Glu Arg Ser Met His Tyr
290 295 300
Gly Ile Glu Val Arg Val Ile Arg Gly Ile Lys Tyr Glu Asn Ser Ile
305 310 315 320
Ser Ser Lys Val Tyr Val Tyr Asp Gly Leu Tyr Lys Ile Val Asp Trp
325 330 335
Trp Phe Ala Val Gly Lys Ser Gly Phe Gly Val Phe Lys Phe Arg Leu
340 345 350
Val Arg Ile Glu Gly Gln Pro Met Met Gly Ser Ala Val Met Arg Phe
355 360 365
Ala Gln Thr Leu Arg Asn Lys Pro Ser Met Val Arg Pro Thr Gly Tyr
370 375 380
Val Ser Phe Asp Leu Ser Asn Lys Lys Glu Asn Val Pro Val Phe Leu
385 390 395 400
Tyr Asn Asp Val Asp Gly Asp Gln Glu Pro Arg His Tyr Glu Tyr Ile
405 410 415
Ala Lys Ala Val Phe Pro Pro Gly Ile Phe Gly Gln Gly Gly Ile Ser
420 425 430
Arg Thr Gly Cys Glu Cys Lys Leu Ser Cys Thr Asp Asp Cys Leu Cys
435 440 445
Ala Arg Lys Asn Gly Gly Glu Phe Ala Tyr Asp Asp Asn Gly His Leu
450 455 460
Leu Lys Gly Lys His Val Val Phe Glu Cys Gly Glu Phe Cys Thr Cys
465 470 475 480
Gly Pro Ser Cys Lys Ser Arg Val Thr Gln Lys Gly Leu Arg Asn Arg
485 490 495
Leu Glu Val Phe Arg Ser Lys Glu Thr Gly Trp Gly Val Arg Thr Leu
500 505 510
Asp Leu Ile Glu Ala Gly Ala Phe Ile Cys Glu Tyr Ala Gly Val Val
515 520 525
Val Thr Arg Leu Gln Ala Glu Ile Leu Ser Met Asn Gly Asp Val Met
530 535 540
Val Tyr Pro Gly Arg Phe Thr Asp Gln Trp Arg Asn Trp Gly Asp Leu
545 550 555 560
Ser Gln Val Tyr Pro Asp Phe Val Arg Pro Asn Tyr Pro Ser Leu Pro
565 570 575
Pro Leu Asp Phe Ser Met Asp Val Ser Arg Met Arg Asn Val Ala Cys
580 585 590
Tyr Ile Ser His Ser Lys Glu Pro Asn Val Met Val Gln Phe Val Leu
595 600 605
His Asp His Asn His Leu Met Phe Pro Arg Val Met Leu Phe Ala Leu
610 615 620
Glu Asn Ile Ser Pro Leu Ala Glu Leu Ser Leu Asp Tyr Gly Leu Ala
625 630 635 640
Asp Glu Val Asn Gly Lys Leu Ala Ile Cys Asn
645 650
<210> 13
<211> 3330
<212> DNA
<213> IR-vTALE-MCS核苷酸序列
<400> 13
atggacccca ttcgttcgcg cacgccaagt cctgcccgcg agcttctgcc cggaccccaa 60
ccggataggg ttcagccgac tgcagatcgg gggggggctc cgcctgctgg cggccccctg 120
gatggcttgc ccgctcggcg gacgatgtcc cggacccggc tgccatctcc ccctgcgccc 180
tcgcctgcgt tctcggcggg cagcttcaac gatctgctcc gtcagttcga tccgtcgctt 240
cttgatacat cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc 300
ccagcagagt gggatgaggt gcaatcgggt ctgcgtgcag ccgatgaccc gccacccacc 360
gtgcgtgtcg ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt 420
gcggcgcaac cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac 480
agtcagcagc agcaagagaa gatcaaatcg aaggtgcgtt cgacagtggc gcagcaccac 540
gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg 600
gcagcgttag ggaccgtcgc tgtcaagtat cagcacataa tcacggcgtt gccagaggcg 660
acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc 720
ttgctcacga aggcggggga gttgagaggt ccgccgttac agttggacac aggccaactt 780
ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc atcgcgcaat 840
gcactgacgg gtgcccccct gaaccttact ccagatcaag ttgtggcaat cgcttcaaat 900
atcggaggga aacaggcctt ggaaaccgtg caacgtcttc ttcctgtgtt gtgtcaagct 960
catggactca ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 1020
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggcct cactccagat 1080
caagttgtgg ctatcgctag taacggtggt ggaaaacagg cgttagaaac tgtccaaagg 1140
ctccttcccg tgttgtgtca agctcatgga ttaactccag atcaagttgt ggcaatcgct 1200
tcaaatatcg gagggaaaca ggccttggaa accgtgcaac gtcttcttcc tgtgttgtgt 1260
caagctcatg gcttaactcc agatcaagtt gtggctatcg ctagtaacgg tggtggaaaa 1320
caggcgttag aaactgtcca aaggctcctt cccgtgttgt gtcaagctca tggactcact 1380
ccagatcaag ttgtggctat cgctagtaac ggtggtggaa aacaggcgtt agaaactgtc 1440
caaaggctcc ttcccgtgtt gtgtcaagct catggcctca ctccagatca agttgtggca 1500
atcgcttcaa atatcggagg gaaacaggcc ttggaaaccg tgcaacgtct tcttcctgtg 1560
ttgtgtcaag ctcatggatt aactccagat caagttgtgg ccattgcatc tcatgatggt 1620
ggtaagcaag ctctcgaaac agtccaaaga cttctcccag tgttgtgtca agctcatggg 1680
ctcactccag atcaagttgt ggccattgca tctcatgatg gtggtaagca agctctcgaa 1740
acagtccaaa gacttctccc agtgttgtgt caagctcatg gactcactcc agatcaagtt 1800
gtggcaatag cctctaacaa tggaggtaaa caagcacttg agactgttca gagattgtta 1860
ccggtgttgt gtcaagctca tggcctcact ccagatcaag ttgtggcaat agcctctaac 1920
aatggaggta aacaagcact tgagactgtt cagagattgt taccggtgtt gtgtcaagct 1980
catggattaa ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 2040
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggtct tactccagat 2100
caagttgtgg ctatcgctag taacggtggt ggaaaacagg cgttagaaac tgtccaaagg 2160
ctccttcccg tgttgtgtca agctcatgga ctcactccag atcaagttgt ggcaatagcc 2220
tctaacaatg gaggtaaaca agcacttgag actgttcaga gattgttacc ggtgttgtgt 2280
caagctcatg gcctcactcc agatcaagtt gtggcaatag cctctaacaa tggaggtaaa 2340
caagcacttg agactgttca gagattgtta ccggtgttgt gtcaagctca tggattaact 2400
ccagatcaag ttgtggccat tgcatctcat gatggtggta agcaagctct cgaaacagtc 2460
caaagacttc tcccagtgtt gtgtcaagct catggactga ccccggacca ggtcgtggcc 2520
atcgccagcc atgatggcgg caagcaggcg ctggagagca ttgttgccca gttatctcgc 2580
cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc 2640
ggacgtcctg ccctggatgc agtgaaaaag ggattgccgc acgcgccgga attgatcaga 2700
agaatcaatc gccgtattcc cgaacgcacg tcccatcgcg ttgccgacct cgcgcacgtg 2760
gtgcgcgtgc ttggtttttt ccagagccac tcccacccag cgcaagcatt cgatgacgcc 2820
atgacgcagt tcgggatgag caggcacggg ttggtacagc tctttcgcag agtgggcgtc 2880
accgaattcg aagcccgctg cggaacgctc cccccagcct cgcagcgttg ggaccgtatc 2940
ctccaggcat cagggatgaa aagggccaaa ccgtccccta cttcagctca aacgccggat 3000
caggcgtctt tgcatgcatt cgccgattcg ctggagcgtg accttgatgc gcccagccca 3060
atgcacgagg gagatcagac gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct 3120
gtcaccggcc cctccacaca gcaatctttc gaggtgcgcg ttcccgaaca gcgcgatgcg 3180
ctgcatttgc ccctcagctg gagggtaaaa cgcccgcgta ccaggatcgg gggcggcctc 3240
ccggatcctg gtacgcccat cgctgccgac ctggcagcgt ccagcaccgt gatgtggtct 3300
agattaatta accccggggt cgactagtga 3330
<210> 14
<211> 5175
<212> DNA
<213> IR-vTALE-AtKYP核苷酸序列
<400> 14
atggacccca ttcgttcgcg cacgccaagt cctgcccgcg agcttctgcc cggaccccaa 60
ccggataggg ttcagccgac tgcagatcgg gggggggctc cgcctgctgg cggccccctg 120
gatggcttgc ccgctcggcg gacgatgtcc cggacccggc tgccatctcc ccctgcgccc 180
tcgcctgcgt tctcggcggg cagcttcaac gatctgctcc gtcagttcga tccgtcgctt 240
cttgatacat cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc 300
ccagcagagt gggatgaggt gcaatcgggt ctgcgtgcag ccgatgaccc gccacccacc 360
gtgcgtgtcg ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt 420
gcggcgcaac cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac 480
agtcagcagc agcaagagaa gatcaaatcg aaggtgcgtt cgacagtggc gcagcaccac 540
gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg 600
gcagcgttag ggaccgtcgc tgtcaagtat cagcacataa tcacggcgtt gccagaggcg 660
acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc 720
ttgctcacga aggcggggga gttgagaggt ccgccgttac agttggacac aggccaactt 780
ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc atcgcgcaat 840
gcactgacgg gtgcccccct gaaccttact ccagatcaag ttgtggcaat cgcttcaaat 900
atcggaggga aacaggcctt ggaaaccgtg caacgtcttc ttcctgtgtt gtgtcaagct 960
catggactca ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 1020
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggcct cactccagat 1080
caagttgtgg ctatcgctag taacggtggt ggaaaacagg cgttagaaac tgtccaaagg 1140
ctccttcccg tgttgtgtca agctcatgga ttaactccag atcaagttgt ggcaatcgct 1200
tcaaatatcg gagggaaaca ggccttggaa accgtgcaac gtcttcttcc tgtgttgtgt 1260
caagctcatg gcttaactcc agatcaagtt gtggctatcg ctagtaacgg tggtggaaaa 1320
caggcgttag aaactgtcca aaggctcctt cccgtgttgt gtcaagctca tggactcact 1380
ccagatcaag ttgtggctat cgctagtaac ggtggtggaa aacaggcgtt agaaactgtc 1440
caaaggctcc ttcccgtgtt gtgtcaagct catggcctca ctccagatca agttgtggca 1500
atcgcttcaa atatcggagg gaaacaggcc ttggaaaccg tgcaacgtct tcttcctgtg 1560
ttgtgtcaag ctcatggatt aactccagat caagttgtgg ccattgcatc tcatgatggt 1620
ggtaagcaag ctctcgaaac agtccaaaga cttctcccag tgttgtgtca agctcatggg 1680
ctcactccag atcaagttgt ggccattgca tctcatgatg gtggtaagca agctctcgaa 1740
acagtccaaa gacttctccc agtgttgtgt caagctcatg gactcactcc agatcaagtt 1800
gtggcaatag cctctaacaa tggaggtaaa caagcacttg agactgttca gagattgtta 1860
ccggtgttgt gtcaagctca tggcctcact ccagatcaag ttgtggcaat agcctctaac 1920
aatggaggta aacaagcact tgagactgtt cagagattgt taccggtgtt gtgtcaagct 1980
catggattaa ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 2040
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggtct tactccagat 2100
caagttgtgg ctatcgctag taacggtggt ggaaaacagg cgttagaaac tgtccaaagg 2160
ctccttcccg tgttgtgtca agctcatgga ctcactccag atcaagttgt ggcaatagcc 2220
tctaacaatg gaggtaaaca agcacttgag actgttcaga gattgttacc ggtgttgtgt 2280
caagctcatg gcctcactcc agatcaagtt gtggcaatag cctctaacaa tggaggtaaa 2340
caagcacttg agactgttca gagattgtta ccggtgttgt gtcaagctca tggattaact 2400
ccagatcaag ttgtggccat tgcatctcat gatggtggta agcaagctct cgaaacagtc 2460
caaagacttc tcccagtgtt gtgtcaagct catggactga ccccggacca ggtcgtggcc 2520
atcgccagcc atgatggcgg caagcaggcg ctggagagca ttgttgccca gttatctcgc 2580
cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc 2640
ggacgtcctg ccctggatgc agtgaaaaag ggattgccgc acgcgccgga attgatcaga 2700
agaatcaatc gccgtattcc cgaacgcacg tcccatcgcg ttgccgacct cgcgcacgtg 2760
gtgcgcgtgc ttggtttttt ccagagccac tcccacccag cgcaagcatt cgatgacgcc 2820
atgacgcagt tcgggatgag caggcacggg ttggtacagc tctttcgcag agtgggcgtc 2880
accgaattcg aagcccgctg cggaacgctc cccccagcct cgcagcgttg ggaccgtatc 2940
ctccaggcat cagggatgaa aagggccaaa ccgtccccta cttcagctca aacgccggat 3000
caggcgtctt tgcatgcatt cgccgattcg ctggagcgtg accttgatgc gcccagccca 3060
atgcacgagg gagatcagac gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct 3120
gtcaccggcc cctccacaca gcaatctttc gaggtgcgcg ttcccgaaca gcgcgatgcg 3180
ctgcatttgc ccctcagctg gagggtaaaa cgcccgcgta ccaggatcgg gggcggcctc 3240
ccggatcctg gtacgcccat cgctgccgac ctggcagcgt ccagcaccgt gatgtggtct 3300
agtgctggaa aaaggaaacg agctaatgct cctgaccaaa cagagcgaag atcgagtgtt 3360
cgggttcaga aagtgagaca gaaagcgtta gatgagaagg cgcgtttagt acaggagagg 3420
gttaagctcc tcagtgacag aaagagtgaa atttgtgtcg atgacactga gttacatgag 3480
aaagaagagg aaaatgtcga tgggagccct aaacgaagaa gccctccaaa gctaaccgca 3540
atgcagaaag gaaagcagaa attgagtgtt tctctgaatg gtaaggacgt gaacttggaa 3600
cctcatctca aagtgacaaa gtgtctgagg ttatttaaca agcaatatct cctctgtgtc 3660
caggctaagt tgagcaggcc tgatttgaag ggtgtaactg agatgataaa agctaaggcg 3720
atattgtacc caagaaaaat aatcggtgac cttccaggta tagacgttgg acaccgtttt 3780
ttttcaagag ctgaaatgtg tgctgtagga ttccataacc attggctaaa tggcattgat 3840
tatatgtcaa tggaatacga aaaagagtat agtaactaca aattaccgct tgctgtttct 3900
attgttatgt cgggccagta cgaggatgat ctagacaatg cagatacagt gacttacact 3960
ggacagggag ggcataactt aactggtaat aaacgtcaga taaaggatca acttttagaa 4020
cgagggaatt tggcgctaaa gcactgctgc gaatataatg tgcctgtcag agtaactcgt 4080
ggtcacaatt gcaaaagtag ctataccaaa cgagtataca cttatgatgg actgtacaag 4140
gttgaaaagt tctgggcaca aaagggcgtt tcaggattta cagtgtataa gtaccgactg 4200
aaacgattgg aggggcaacc agaactaact actgatcagg tcaactttgt tgctggacgc 4260
ataccaacga gtacttcaga aattgagggt ttggtatgtg aggacatctc cggagggcta 4320
gaatttaagg gtatccccgc cactaatcgt gttgatgatt caccagtttc accaacatct 4380
ggtttcacat acatcaaatc tttgattatt gagcctaatg tcataattcc aaagagttca 4440
actgggtgta actgccgagg cagctgcact gactcaaaga aatgtgcatg tgctaagctt 4500
aatgggggta actttccata tgttgacctt aatgatggca gattaattga gtctcgagat 4560
gttgtatttg aatgtggtcc tcactgtggg tgtgggccaa aatgtgtcaa ccgaacttct 4620
cagaagcgtc taagattcaa tcttgaggtt ttccgctctg caaagaaggg ttgggcagtt 4680
agatcatggg agtacatacc agctggttca ccagtatgtg agtacatagg agttgtcagg 4740
agaactgctg atgtggatac tatctctgac aatgaataca tatttgagat tgactgccaa 4800
cagacaatgc aaggtcttgg tggaagacag agaagactaa gagatgttgc tgtaccaatg 4860
aataatggag tcagtcagag cagtgaagat gagaatgcgc cagagttctg cattgatgct 4920
ggttcaacag gaaactttgc taggtttata aatcacagtt gtgaaccaaa cctatttgtt 4980
cagtgcgtcc tgagttctca ccaggatata aggcttgccc gtgtggttct tttcgcagct 5040
gacaacattt ccccaatgca ggagctcact tacgactatg gatatgcgct tgatagcgtt 5100
catggaccgg atgggaaggt gaagcagctc gcttgctact gtggagcgct aaattgtagg 5160
aaacgccttt actaa 5175
<210> 15
<211> 4710
<212> DNA
<213> IR-vTALE-AtHDAC19核苷酸序列
<400> 15
atggacccca ttcgttcgcg cacgccaagt cctgcccgcg agcttctgcc cggaccccaa 60
ccggataggg ttcagccgac tgcagatcgg gggggggctc cgcctgctgg cggccccctg 120
gatggcttgc ccgctcggcg gacgatgtcc cggacccggc tgccatctcc ccctgcgccc 180
tcgcctgcgt tctcggcggg cagcttcaac gatctgctcc gtcagttcga tccgtcgctt 240
cttgatacat cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc 300
ccagcagagt gggatgaggt gcaatcgggt ctgcgtgcag ccgatgaccc gccacccacc 360
gtgcgtgtcg ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt 420
gcggcgcaac cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac 480
agtcagcagc agcaagagaa gatcaaatcg aaggtgcgtt cgacagtggc gcagcaccac 540
gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg 600
gcagcgttag ggaccgtcgc tgtcaagtat cagcacataa tcacggcgtt gccagaggcg 660
acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc 720
ttgctcacga aggcggggga gttgagaggt ccgccgttac agttggacac aggccaactt 780
ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc atcgcgcaat 840
gcactgacgg gtgcccccct gaaccttact ccagatcaag ttgtggcaat cgcttcaaat 900
atcggaggga aacaggcctt ggaaaccgtg caacgtcttc ttcctgtgtt gtgtcaagct 960
catggactca ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 1020
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggcct cactccagat 1080
caagttgtgg ctatcgctag taacggtggt ggaaaacagg cgttagaaac tgtccaaagg 1140
ctccttcccg tgttgtgtca agctcatgga ttaactccag atcaagttgt ggcaatcgct 1200
tcaaatatcg gagggaaaca ggccttggaa accgtgcaac gtcttcttcc tgtgttgtgt 1260
caagctcatg gcttaactcc agatcaagtt gtggctatcg ctagtaacgg tggtggaaaa 1320
caggcgttag aaactgtcca aaggctcctt cccgtgttgt gtcaagctca tggactcact 1380
ccagatcaag ttgtggctat cgctagtaac ggtggtggaa aacaggcgtt agaaactgtc 1440
caaaggctcc ttcccgtgtt gtgtcaagct catggcctca ctccagatca agttgtggca 1500
atcgcttcaa atatcggagg gaaacaggcc ttggaaaccg tgcaacgtct tcttcctgtg 1560
ttgtgtcaag ctcatggatt aactccagat caagttgtgg ccattgcatc tcatgatggt 1620
ggtaagcaag ctctcgaaac agtccaaaga cttctcccag tgttgtgtca agctcatggg 1680
ctcactccag atcaagttgt ggccattgca tctcatgatg gtggtaagca agctctcgaa 1740
acagtccaaa gacttctccc agtgttgtgt caagctcatg gactcactcc agatcaagtt 1800
gtggcaatag cctctaacaa tggaggtaaa caagcacttg agactgttca gagattgtta 1860
ccggtgttgt gtcaagctca tggcctcact ccagatcaag ttgtggcaat agcctctaac 1920
aatggaggta aacaagcact tgagactgtt cagagattgt taccggtgtt gtgtcaagct 1980
catggattaa ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 2040
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggtct tactccagat 2100
caagttgtgg ctatcgctag taacggtggt ggaaaacagg cgttagaaac tgtccaaagg 2160
ctccttcccg tgttgtgtca agctcatgga ctcactccag atcaagttgt ggcaatagcc 2220
tctaacaatg gaggtaaaca agcacttgag actgttcaga gattgttacc ggtgttgtgt 2280
caagctcatg gcctcactcc agatcaagtt gtggcaatag cctctaacaa tggaggtaaa 2340
caagcacttg agactgttca gagattgtta ccggtgttgt gtcaagctca tggattaact 2400
ccagatcaag ttgtggccat tgcatctcat gatggtggta agcaagctct cgaaacagtc 2460
caaagacttc tcccagtgtt gtgtcaagct catggactga ccccggacca ggtcgtggcc 2520
atcgccagcc atgatggcgg caagcaggcg ctggagagca ttgttgccca gttatctcgc 2580
cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc 2640
ggacgtcctg ccctggatgc agtgaaaaag ggattgccgc acgcgccgga attgatcaga 2700
agaatcaatc gccgtattcc cgaacgcacg tcccatcgcg ttgccgacct cgcgcacgtg 2760
gtgcgcgtgc ttggtttttt ccagagccac tcccacccag cgcaagcatt cgatgacgcc 2820
atgacgcagt tcgggatgag caggcacggg ttggtacagc tctttcgcag agtgggcgtc 2880
accgaattcg aagcccgctg cggaacgctc cccccagcct cgcagcgttg ggaccgtatc 2940
ctccaggcat cagggatgaa aagggccaaa ccgtccccta cttcagctca aacgccggat 3000
caggcgtctt tgcatgcatt cgccgattcg ctggagcgtg accttgatgc gcccagccca 3060
atgcacgagg gagatcagac gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct 3120
gtcaccggcc cctccacaca gcaatctttc gaggtgcgcg ttcccgaaca gcgcgatgcg 3180
ctgcatttgc ccctcagctg gagggtaaaa cgcccgcgta ccaggatcgg gggcggcctc 3240
ccggatcctg gtacgcccat cgctgccgac ctggcagcgt ccagcaccgt gatgtggtct 3300
agtgatactg gcggcaattc gctggcgtcc ggacctgatg gtgtgaagag gaaagtttgt 3360
tatttctatg accctgaggt cggcaattac tactatggcc aaggtcatcc catgaagccc 3420
catcgcatcc gcatgaccca tgccctcctc gctcactacg gtctccttca gcatatgcag 3480
gttctcaagc ccttccctgc ccgcgaccgt gatctctgcc gcttccacgc cgacgactat 3540
gtctcttttc tccgcagcat tacccctgaa acccagcaag atcagattcg ccaacttaag 3600
cgcttcaatg ttggtgaaga ctgtcccgtc tttgacggcc tttattcctt ttgccagacc 3660
tatgctggag gatctgttgg tggctctgtc aagcttaacc acggcctctg cgatattgcc 3720
atcaactggg ctggtggtct ccatcacgct aagaagtgcg aggcctctgg cttctgttac 3780
gtcaatgata tcgtcttagc tatcctagag ctccttaagc agcatgagcg tgttctttat 3840
gtcgatattg atatccacca cggggatgga gtggaggagg cattttatgc tactgacagg 3900
gttatgactg tctcgtttca taaatttggt gattactttc ccggtacagg tcacattcag 3960
gatataggtt atggtagcgg aaagtactat tctctcaatg taccactgga tgatggaatc 4020
gatgatgaga gctatcatct gttattcaag cccatcatgg ggaaagttat ggaaattttc 4080
cgaccagggg ctgtggtatt gcaatgtggt gctgattcat tgtctggtga taggttgggg 4140
tgctttaatc tttcaatcaa aggtcatgct gagtgcgtca aatttatgag atcgttcaat 4200
gttcccctac tgctcttggg tggtggtggt tacactatcc gcaatgttgc ccgttgctgg 4260
tgctacgaga ctggagttgc acttggagtt gaagttgaag acaagatgcc ggagcatgaa 4320
tattatgaat actttggtcc agactataca cttcacgttg ctccaagtaa catggaaaat 4380
aagaattctc gtcagatgct tgaagagatt cgcaatgacc ttctccacaa tctctctaag 4440
cttcagcatg ctccaagtgt accatttcag gaaagaccac ctgatacaga gactcccgag 4500
gttgatgaag accaagaaga tggggataaa agatgggatc cggattcaga catggatgtt 4560
gatgatgacc gtaaacctat accaagcaga gtaaaaagag aagctgttga accagataca 4620
aaggacaagg atggactgaa aggaattatg gagcgtggaa aaggttgtga ggtggaggtg 4680
gatgagagtg gaagcactaa ggtaaaatga 4710
<210> 16
<211> 7905
<212> DNA
<213> IR-vTALE-AtMET1核苷酸序列
<400> 16
atggacccca ttcgttcgcg cacgccaagt cctgcccgcg agcttctgcc cggaccccaa 60
ccggataggg ttcagccgac tgcagatcgg gggggggctc cgcctgctgg cggccccctg 120
gatggcttgc ccgctcggcg gacgatgtcc cggacccggc tgccatctcc ccctgcgccc 180
tcgcctgcgt tctcggcggg cagcttcaac gatctgctcc gtcagttcga tccgtcgctt 240
cttgatacat cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc 300
ccagcagagt gggatgaggt gcaatcgggt ctgcgtgcag ccgatgaccc gccacccacc 360
gtgcgtgtcg ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt 420
gcggcgcaac cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac 480
agtcagcagc agcaagagaa gatcaaatcg aaggtgcgtt cgacagtggc gcagcaccac 540
gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg 600
gcagcgttag ggaccgtcgc tgtcaagtat cagcacataa tcacggcgtt gccagaggcg 660
acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc 720
ttgctcacga aggcggggga gttgagaggt ccgccgttac agttggacac aggccaactt 780
ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc atcgcgcaat 840
gcactgacgg gtgcccccct gaaccttact ccagatcaag ttgtggcaat cgcttcaaat 900
atcggaggga aacaggcctt ggaaaccgtg caacgtcttc ttcctgtgtt gtgtcaagct 960
catggactca ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 1020
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggcct cactccagat 1080
caagttgtgg ctatcgctag taacggtggt ggaaaacagg cgttagaaac tgtccaaagg 1140
ctccttcccg tgttgtgtca agctcatgga ttaactccag atcaagttgt ggcaatcgct 1200
tcaaatatcg gagggaaaca ggccttggaa accgtgcaac gtcttcttcc tgtgttgtgt 1260
caagctcatg gcttaactcc agatcaagtt gtggctatcg ctagtaacgg tggtggaaaa 1320
caggcgttag aaactgtcca aaggctcctt cccgtgttgt gtcaagctca tggactcact 1380
ccagatcaag ttgtggctat cgctagtaac ggtggtggaa aacaggcgtt agaaactgtc 1440
caaaggctcc ttcccgtgtt gtgtcaagct catggcctca ctccagatca agttgtggca 1500
atcgcttcaa atatcggagg gaaacaggcc ttggaaaccg tgcaacgtct tcttcctgtg 1560
ttgtgtcaag ctcatggatt aactccagat caagttgtgg ccattgcatc tcatgatggt 1620
ggtaagcaag ctctcgaaac agtccaaaga cttctcccag tgttgtgtca agctcatggg 1680
ctcactccag atcaagttgt ggccattgca tctcatgatg gtggtaagca agctctcgaa 1740
acagtccaaa gacttctccc agtgttgtgt caagctcatg gactcactcc agatcaagtt 1800
gtggcaatag cctctaacaa tggaggtaaa caagcacttg agactgttca gagattgtta 1860
ccggtgttgt gtcaagctca tggcctcact ccagatcaag ttgtggcaat agcctctaac 1920
aatggaggta aacaagcact tgagactgtt cagagattgt taccggtgtt gtgtcaagct 1980
catggattaa ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 2040
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggtct tactccagat 2100
caagttgtgg ctatcgctag taacggtggt ggaaaacagg cgttagaaac tgtccaaagg 2160
ctccttcccg tgttgtgtca agctcatgga ctcactccag atcaagttgt ggcaatagcc 2220
tctaacaatg gaggtaaaca agcacttgag actgttcaga gattgttacc ggtgttgtgt 2280
caagctcatg gcctcactcc agatcaagtt gtggcaatag cctctaacaa tggaggtaaa 2340
caagcacttg agactgttca gagattgtta ccggtgttgt gtcaagctca tggattaact 2400
ccagatcaag ttgtggccat tgcatctcat gatggtggta agcaagctct cgaaacagtc 2460
caaagacttc tcccagtgtt gtgtcaagct catggactga ccccggacca ggtcgtggcc 2520
atcgccagcc atgatggcgg caagcaggcg ctggagagca ttgttgccca gttatctcgc 2580
cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc 2640
ggacgtcctg ccctggatgc agtgaaaaag ggattgccgc acgcgccgga attgatcaga 2700
agaatcaatc gccgtattcc cgaacgcacg tcccatcgcg ttgccgacct cgcgcacgtg 2760
gtgcgcgtgc ttggtttttt ccagagccac tcccacccag cgcaagcatt cgatgacgcc 2820
atgacgcagt tcgggatgag caggcacggg ttggtacagc tctttcgcag agtgggcgtc 2880
accgaattcg aagcccgctg cggaacgctc cccccagcct cgcagcgttg ggaccgtatc 2940
ctccaggcat cagggatgaa aagggccaaa ccgtccccta cttcagctca aacgccggat 3000
caggcgtctt tgcatgcatt cgccgattcg ctggagcgtg accttgatgc gcccagccca 3060
atgcacgagg gagatcagac gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct 3120
gtcaccggcc cctccacaca gcaatctttc gaggtgcgcg ttcccgaaca gcgcgatgcg 3180
ctgcatttgc ccctcagctg gagggtaaaa cgcccgcgta ccaggatcgg gggcggcctc 3240
ccggatcctg gtacgcccat cgctgccgac ctggcagcgt ccagcaccgt gatgtggtct 3300
agtgtggaaa atggggctaa agctgcgaag cgaaagaaga gaccacttcc agagattcaa 3360
gaggtagaag atgtacctag gacgaggaga ccaaggcgtg ctgcagcgtg taccagtttc 3420
aaggagaaat ctattcgagt ctgtgagaaa tctgctacta ttgaagtaaa gaaacagcag 3480
attgtggagg aagagtttct cgcgttacgg ttaacggctc tggaaactga tgttgaagat 3540
cgtccaacca ggagactgaa tgattttgtt ttgtttgatt cagatggagt tccacaacct 3600
ctggagatgt tggagattca tgacatattc gtttcaggtg ctatcttacc ttcagatgtg 3660
tgtactgata aggagaaaga gaagggtgtg aggtgtacat cgtttggacg ggttgagcat 3720
tggagtatct ctggttatga agatggttcc cctgttattt ggatctcaac ggaattggcg 3780
gattatgatt gtcgtaaacc tgctgctagc tacaggaagg tttatgatta cttctatgag 3840
aaagctcgtg cttcagtggc tgtgtataag aaattgtcca agtcatctgg tggggatcct 3900
gatataggtc ttgaggagtt acttgcggcg gttgtcagat caatgagcag tggaagcaag 3960
tacttttcta gtggtgcggc aatcatcgat tttgttatat cccagggaga ttttatatat 4020
aaccaactcg ctggtttgga tgagacagcc aagaaacatg aatcaagcta tgttgagatt 4080
cctgttcttg tagctctcag agagaagagt agtaagattg acaagcctct gcagagggaa 4140
agaaacccat ctaatggtgt gaggattaaa gaagtttctc aagttgcgga gagcgaggcc 4200
ttgacatctg atcaactggt tgatggtact gatgatgaca gaagatatgc tatactctta 4260
caagacgaag agaataggaa atctatgcaa cagcccagaa aaaacagcag ctcaggttct 4320
gcttcaaata tgttctacat taagataaat gaagatgaga ttgccaatga ttatcctctc 4380
ccatcgtact ataagacctc cgaagaagaa acagatgaac ttatacttta tgatgcttcc 4440
tatgaggttc aatctgaaca cctgcctcac aggatgcttc acaactgggc tctttataac 4500
tctgatttac gattcatatc actggaactt ctaccgatga aacaatgtga tgatattgat 4560
gtcaacattt ttgggtcagg tgtggtgact gatgataatg gaagttggat ttctttaaac 4620
gatcctgaca gcggttctca gtcacacgat cctgatggga tgtgcatatt cctcagtcaa 4680
attaaagaat ggatgattga gtttgggagc gatgatatta tctccatttc tatacgaaca 4740
gatgtggcct ggtaccgtct tgggaaacca tcaaaacttt atgccccttg gtggaaacct 4800
gttctgaaaa cagcaagggt tgggataagc attcttactt ttcttagggt ggaaagtagg 4860
gttgctaggc tttcatttgc agatgtcaca aaaagactgt ctgggttaca ggcgaatgat 4920
aaagcttaca tttcttctga ccccttggct gttgagagat atttggtcgt ccatgggcaa 4980
attattttac agctttttgc agtttatccg gacgacaatg tcaaaaggtg tccatttgtt 5040
gttggtcttg caagcaaatt ggaggatagg caccacacaa aatggatcat caagaagaag 5100
aaaatttcgc tgaaggaact gaatctgaat ccaagggcag gcatggcacc agtagcatcg 5160
aagaggaaag ctatgcaagc aacaacaact cgcctggtca acagaatttg gggagagttt 5220
tactccaatt actctccaga ggatccattg caggcgactg ctgcagaaaa tggggaggat 5280
gaggtggaag aggaaggcgg aaatggggag gaagaggttg aagaggaagg tgaaaatggt 5340
ctcacagagg acactgtacc agaacctgtt gaggttcaga agcctcatac tcctaagaaa 5400
atccgaggca gttctggaaa aagggaaata aaatgggatg gtgagagtct aggaaaaact 5460
tctgctggcg agcctctcta tcaacaagcc cttgttggag gggaaatggt ggctgtaggt 5520
ggcgctgtca ccttggaagt tgatgatcca gatgaaatgc cggccatcta ttttgtggag 5580
tacatgttcg aaagtacaga tcactgcaaa atgttacatg gtagattctt acaaagagga 5640
tctatgactg ttctggggaa tgctgctaac gagagggaac tattcctgac taatgaatgc 5700
atgactacac agctcaagga cattaaagga gtagccagtt ttgagattcg atcaaggcca 5760
tgggggcatc agtataggaa aaagaacatc actgcggata agcttgactg ggctagagca 5820
ttagaaagaa aagtaaaaga tttgccaaca gagtattact gcaaaagctt gtactcacct 5880
gagagagggg gattctttag tcttccacta agtgatattg gtcgcagttc tgggttctgc 5940
acttcatgta agataaggga ggatgaagag aagaggtcta caattaaact aaatgtttca 6000
aagacaggct ttttcatcaa tgggattgag tattctgttg aggattttgt ctatgtcaac 6060
cctgactcta ttggtgggtt gaaggagggt agtaaaactt cttttaagtc tgggcgaaac 6120
attgggttaa gagcgtatgt tgtttgccaa ttgctggaaa ttgttccaaa ggaatctaga 6180
aaggctgatt tgggttcctt tgatgttaaa gtgagaaggt tttataggcc tgaggatgtt 6240
tctgcagaga aggcctatgc ttcagacatc caagaattgt atttcagcca ggacacagtt 6300
gttctccctc caggtgctct agagggaaaa tgtgaagtaa gaaagaaaag tgatatgccc 6360
ttatcccgtg aatatccaat atcagaccat attttcttct gtgatctttt ctttgacacc 6420
tccaaaggtt ctctcaagca gctgcccgcc aatatgaagc caaagttctc tactattaag 6480
gacgacacac ttttaagaaa gaaaaaggga aagggagtag agagtgaaat tgagtctgag 6540
attgtcaagc ctgttgagcc acctaaagag attcgtctgg ctactctaga tatttttgct 6600
ggttgtggtg gcctgtctca tggactgaaa aaggcgggtg tatctgatgc aaagtgggcg 6660
attgagtatg aagagccagc tgggcaggct tttaaacaaa accatcctga gtcaacagtt 6720
tttgttgaca actgcaatgt gattcttagg gctataatgg agaaaggtgg agatcaagat 6780
gattgtgtct ctactacaga ggcaaatgaa ttagcagcta aactaactga ggagcagaag 6840
agtactctgc cactgcctgg tcaagtggac ttcatcaatg gtggacctcc atgtcaggga 6900
ttttctggta tgaacaggtt caaccaaagc tcttggagta aagttcagtg tgaaatgata 6960
ttagcattct tgtcctttgc tgactatttc cggccaaggt attttcttct ggagaacgtg 7020
aggacctttg tgtcattcaa taaagggcag acatttcagc ttactttggc ttcccttctc 7080
gaaatgggtt accaggtgag atttggaatc ctggaggccg gtgcatatgg agtatcccaa 7140
tctcgtaaac gagctttcat ttgggctgct gcaccagaag aagttctccc tgaatggcct 7200
gagccgatgc atgtctttgg tgttccaaag ttgaaaatct cactatctca aggtttacat 7260
tatgctgctg ttcgtagtac tgcacttggt gcccctttcc gtccaatcac cgtgagagac 7320
acaattggtg atcttccatc agtagaaaac ggagactcta ggacaaacaa agagtataaa 7380
gaggttgcag tctcgtggtt ccaaaaggag ataagaggaa acacgattgc tctcactgat 7440
catatctgca aggctatgaa tgagcttaac ctcattcgat gcaaattaat cccaactagg 7500
cctggggctg attggcatga cttgccaaag agaaaggtta cgttatctga tgggcgcgta 7560
gaagaaatga ttcctttttg tctcccaaac acagctgagc gccacaacgg ttggaaggga 7620
ctatatggga gattagattg gcaaggaaac tttccgactt ccgtcacgga tcctcagccc 7680
atgggtaagg ttggaatgtg ctttcatcct gaacagcaca gaatccttac agtccgtgaa 7740
tgcgcccgat ctcaggggtt tccggatagc tacgagtttg cagggaacat aaatcacaag 7800
cacaggcaga ttgggaatgc agtccctcca ccattggcat ttgctctagg tcgtaagctc 7860
aaagaagccc tacatctcaa gaagtctcct caacaccaac cctag 7905
<210> 17
<211> 1724
<212> PRT
<213> IR-vTALE-AtKYP氨基酸序列
<400> 17
Met Asp Pro Ile Arg Ser Arg Thr Pro Ser Pro Ala Arg Glu Leu Leu
1 5 10 15
Pro Gly Pro Gln Pro Asp Arg Val Gln Pro Thr Ala Asp Arg Gly Gly
20 25 30
Ala Pro Pro Ala Gly Gly Pro Leu Asp Gly Leu Pro Ala Arg Arg Thr
35 40 45
Met Ser Arg Thr Arg Leu Pro Ser Pro Pro Ala Pro Ser Pro Ala Phe
50 55 60
Ser Ala Gly Ser Phe Asn Asp Leu Leu Arg Gln Phe Asp Pro Ser Leu
65 70 75 80
Leu Asp Thr Ser Leu Leu Asp Ser Met Pro Ala Val Gly Thr Pro His
85 90 95
Thr Ala Ala Ala Pro Ala Glu Trp Asp Glu Val Gln Ser Gly Leu Arg
100 105 110
Ala Ala Asp Asp Pro Pro Pro Thr Val Arg Val Ala Val Thr Ala Ala
115 120 125
Arg Pro Pro Arg Ala Lys Pro Ala Pro Arg Arg Arg Ala Ala Gln Pro
130 135 140
Ser Asp Ala Ser Pro Ala Ala Gln Val Asp Leu Arg Thr Leu Gly Tyr
145 150 155 160
Ser Gln Gln Gln Gln Glu Lys Ile Lys Ser Lys Val Arg Ser Thr Val
165 170 175
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
180 185 190
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
195 200 205
Lys Tyr Gln His Ile Ile Thr Ala Leu Pro Glu Ala Thr His Glu Asp
210 215 220
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
225 230 235 240
Leu Leu Thr Lys Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
245 250 255
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
260 265 270
Glu Ala Val His Ala Ser Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
275 280 285
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys
290 295 300
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
305 310 315 320
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly
325 330 335
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
340 345 350
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn
355 360 365
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
370 375 380
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
385 390 395 400
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
405 410 415
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
420 425 430
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
435 440 445
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
450 455 460
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
465 470 475 480
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
485 490 495
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
500 505 510
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
515 520 525
Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
530 535 540
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
545 550 555 560
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
565 570 575
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
580 585 590
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
595 600 605
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
610 615 620
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn
625 630 635 640
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
645 650 655
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
660 665 670
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
675 680 685
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
690 695 700
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
705 710 715 720
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
725 730 735
Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val
740 745 750
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
755 760 765
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
770 775 780
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
785 790 795 800
Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
805 810 815
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
820 825 830
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
835 840 845
Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala
850 855 860
Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly
865 870 875 880
Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Pro His Ala Pro
885 890 895
Glu Leu Ile Arg Arg Ile Asn Arg Arg Ile Pro Glu Arg Thr Ser His
900 905 910
Arg Val Ala Asp Leu Ala His Val Val Arg Val Leu Gly Phe Phe Gln
915 920 925
Ser His Ser His Pro Ala Gln Ala Phe Asp Asp Ala Met Thr Gln Phe
930 935 940
Gly Met Ser Arg His Gly Leu Val Gln Leu Phe Arg Arg Val Gly Val
945 950 955 960
Thr Glu Phe Glu Ala Arg Cys Gly Thr Leu Pro Pro Ala Ser Gln Arg
965 970 975
Trp Asp Arg Ile Leu Gln Ala Ser Gly Met Lys Arg Ala Lys Pro Ser
980 985 990
Pro Thr Ser Ala Gln Thr Pro Asp Gln Ala Ser Leu His Ala Phe Ala
995 1000 1005
Asp Ser Leu Glu Arg Asp Leu Asp Ala Pro Ser Pro Met His Glu
1010 1015 1020
Gly Asp Gln Thr Arg Ala Ser Ser Arg Lys Arg Ser Arg Ser Asp
1025 1030 1035
Arg Ala Val Thr Gly Pro Ser Thr Gln Gln Ser Phe Glu Val Arg
1040 1045 1050
Val Pro Glu Gln Arg Asp Ala Leu His Leu Pro Leu Ser Trp Arg
1055 1060 1065
Val Lys Arg Pro Arg Thr Arg Ile Gly Gly Gly Leu Pro Asp Pro
1070 1075 1080
Gly Thr Pro Ile Ala Ala Asp Leu Ala Ala Ser Ser Thr Val Met
1085 1090 1095
Trp Ser Ser Ala Gly Lys Arg Lys Arg Ala Asn Ala Pro Asp Gln
1100 1105 1110
Thr Glu Arg Arg Ser Ser Val Arg Val Gln Lys Val Arg Gln Lys
1115 1120 1125
Ala Leu Asp Glu Lys Ala Arg Leu Val Gln Glu Arg Val Lys Leu
1130 1135 1140
Leu Ser Asp Arg Lys Ser Glu Ile Cys Val Asp Asp Thr Glu Leu
1145 1150 1155
His Glu Lys Glu Glu Glu Asn Val Asp Gly Ser Pro Lys Arg Arg
1160 1165 1170
Ser Pro Pro Lys Leu Thr Ala Met Gln Lys Gly Lys Gln Lys Leu
1175 1180 1185
Ser Val Ser Leu Asn Gly Lys Asp Val Asn Leu Glu Pro His Leu
1190 1195 1200
Lys Val Thr Lys Cys Leu Arg Leu Phe Asn Lys Gln Tyr Leu Leu
1205 1210 1215
Cys Val Gln Ala Lys Leu Ser Arg Pro Asp Leu Lys Gly Val Thr
1220 1225 1230
Glu Met Ile Lys Ala Lys Ala Ile Leu Tyr Pro Arg Lys Ile Ile
1235 1240 1245
Gly Asp Leu Pro Gly Ile Asp Val Gly His Arg Phe Phe Ser Arg
1250 1255 1260
Ala Glu Met Cys Ala Val Gly Phe His Asn His Trp Leu Asn Gly
1265 1270 1275
Ile Asp Tyr Met Ser Met Glu Tyr Glu Lys Glu Tyr Ser Asn Tyr
1280 1285 1290
Lys Leu Pro Leu Ala Val Ser Ile Val Met Ser Gly Gln Tyr Glu
1295 1300 1305
Asp Asp Leu Asp Asn Ala Asp Thr Val Thr Tyr Thr Gly Gln Gly
1310 1315 1320
Gly His Asn Leu Thr Gly Asn Lys Arg Gln Ile Lys Asp Gln Leu
1325 1330 1335
Leu Glu Arg Gly Asn Leu Ala Leu Lys His Cys Cys Glu Tyr Asn
1340 1345 1350
Val Pro Val Arg Val Thr Arg Gly His Asn Cys Lys Ser Ser Tyr
1355 1360 1365
Thr Lys Arg Val Tyr Thr Tyr Asp Gly Leu Tyr Lys Val Glu Lys
1370 1375 1380
Phe Trp Ala Gln Lys Gly Val Ser Gly Phe Thr Val Tyr Lys Tyr
1385 1390 1395
Arg Leu Lys Arg Leu Glu Gly Gln Pro Glu Leu Thr Thr Asp Gln
1400 1405 1410
Val Asn Phe Val Ala Gly Arg Ile Pro Thr Ser Thr Ser Glu Ile
1415 1420 1425
Glu Gly Leu Val Cys Glu Asp Ile Ser Gly Gly Leu Glu Phe Lys
1430 1435 1440
Gly Ile Pro Ala Thr Asn Arg Val Asp Asp Ser Pro Val Ser Pro
1445 1450 1455
Thr Ser Gly Phe Thr Tyr Ile Lys Ser Leu Ile Ile Glu Pro Asn
1460 1465 1470
Val Ile Ile Pro Lys Ser Ser Thr Gly Cys Asn Cys Arg Gly Ser
1475 1480 1485
Cys Thr Asp Ser Lys Lys Cys Ala Cys Ala Lys Leu Asn Gly Gly
1490 1495 1500
Asn Phe Pro Tyr Val Asp Leu Asn Asp Gly Arg Leu Ile Glu Ser
1505 1510 1515
Arg Asp Val Val Phe Glu Cys Gly Pro His Cys Gly Cys Gly Pro
1520 1525 1530
Lys Cys Val Asn Arg Thr Ser Gln Lys Arg Leu Arg Phe Asn Leu
1535 1540 1545
Glu Val Phe Arg Ser Ala Lys Lys Gly Trp Ala Val Arg Ser Trp
1550 1555 1560
Glu Tyr Ile Pro Ala Gly Ser Pro Val Cys Glu Tyr Ile Gly Val
1565 1570 1575
Val Arg Arg Thr Ala Asp Val Asp Thr Ile Ser Asp Asn Glu Tyr
1580 1585 1590
Ile Phe Glu Ile Asp Cys Gln Gln Thr Met Gln Gly Leu Gly Gly
1595 1600 1605
Arg Gln Arg Arg Leu Arg Asp Val Ala Val Pro Met Asn Asn Gly
1610 1615 1620
Val Ser Gln Ser Ser Glu Asp Glu Asn Ala Pro Glu Phe Cys Ile
1625 1630 1635
Asp Ala Gly Ser Thr Gly Asn Phe Ala Arg Phe Ile Asn His Ser
1640 1645 1650
Cys Glu Pro Asn Leu Phe Val Gln Cys Val Leu Ser Ser His Gln
1655 1660 1665
Asp Ile Arg Leu Ala Arg Val Val Leu Phe Ala Ala Asp Asn Ile
1670 1675 1680
Ser Pro Met Gln Glu Leu Thr Tyr Asp Tyr Gly Tyr Ala Leu Asp
1685 1690 1695
Ser Val His Gly Pro Asp Gly Lys Val Lys Gln Leu Ala Cys Tyr
1700 1705 1710
Cys Gly Ala Leu Asn Cys Arg Lys Arg Leu Tyr
1715 1720
<210> 18
<211> 1569
<212> PRT
<213> IR-vTALE-AtHDAC19氨基酸序列
<400> 18
Met Asp Pro Ile Arg Ser Arg Thr Pro Ser Pro Ala Arg Glu Leu Leu
1 5 10 15
Pro Gly Pro Gln Pro Asp Arg Val Gln Pro Thr Ala Asp Arg Gly Gly
20 25 30
Ala Pro Pro Ala Gly Gly Pro Leu Asp Gly Leu Pro Ala Arg Arg Thr
35 40 45
Met Ser Arg Thr Arg Leu Pro Ser Pro Pro Ala Pro Ser Pro Ala Phe
50 55 60
Ser Ala Gly Ser Phe Asn Asp Leu Leu Arg Gln Phe Asp Pro Ser Leu
65 70 75 80
Leu Asp Thr Ser Leu Leu Asp Ser Met Pro Ala Val Gly Thr Pro His
85 90 95
Thr Ala Ala Ala Pro Ala Glu Trp Asp Glu Val Gln Ser Gly Leu Arg
100 105 110
Ala Ala Asp Asp Pro Pro Pro Thr Val Arg Val Ala Val Thr Ala Ala
115 120 125
Arg Pro Pro Arg Ala Lys Pro Ala Pro Arg Arg Arg Ala Ala Gln Pro
130 135 140
Ser Asp Ala Ser Pro Ala Ala Gln Val Asp Leu Arg Thr Leu Gly Tyr
145 150 155 160
Ser Gln Gln Gln Gln Glu Lys Ile Lys Ser Lys Val Arg Ser Thr Val
165 170 175
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
180 185 190
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
195 200 205
Lys Tyr Gln His Ile Ile Thr Ala Leu Pro Glu Ala Thr His Glu Asp
210 215 220
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
225 230 235 240
Leu Leu Thr Lys Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
245 250 255
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
260 265 270
Glu Ala Val His Ala Ser Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
275 280 285
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys
290 295 300
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
305 310 315 320
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly
325 330 335
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
340 345 350
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn
355 360 365
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
370 375 380
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
385 390 395 400
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
405 410 415
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
420 425 430
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
435 440 445
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
450 455 460
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
465 470 475 480
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
485 490 495
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
500 505 510
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
515 520 525
Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
530 535 540
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
545 550 555 560
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
565 570 575
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
580 585 590
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
595 600 605
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
610 615 620
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn
625 630 635 640
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
645 650 655
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
660 665 670
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
675 680 685
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
690 695 700
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
705 710 715 720
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
725 730 735
Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val
740 745 750
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
755 760 765
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
770 775 780
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
785 790 795 800
Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
805 810 815
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
820 825 830
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
835 840 845
Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala
850 855 860
Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly
865 870 875 880
Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Pro His Ala Pro
885 890 895
Glu Leu Ile Arg Arg Ile Asn Arg Arg Ile Pro Glu Arg Thr Ser His
900 905 910
Arg Val Ala Asp Leu Ala His Val Val Arg Val Leu Gly Phe Phe Gln
915 920 925
Ser His Ser His Pro Ala Gln Ala Phe Asp Asp Ala Met Thr Gln Phe
930 935 940
Gly Met Ser Arg His Gly Leu Val Gln Leu Phe Arg Arg Val Gly Val
945 950 955 960
Thr Glu Phe Glu Ala Arg Cys Gly Thr Leu Pro Pro Ala Ser Gln Arg
965 970 975
Trp Asp Arg Ile Leu Gln Ala Ser Gly Met Lys Arg Ala Lys Pro Ser
980 985 990
Pro Thr Ser Ala Gln Thr Pro Asp Gln Ala Ser Leu His Ala Phe Ala
995 1000 1005
Asp Ser Leu Glu Arg Asp Leu Asp Ala Pro Ser Pro Met His Glu
1010 1015 1020
Gly Asp Gln Thr Arg Ala Ser Ser Arg Lys Arg Ser Arg Ser Asp
1025 1030 1035
Arg Ala Val Thr Gly Pro Ser Thr Gln Gln Ser Phe Glu Val Arg
1040 1045 1050
Val Pro Glu Gln Arg Asp Ala Leu His Leu Pro Leu Ser Trp Arg
1055 1060 1065
Val Lys Arg Pro Arg Thr Arg Ile Gly Gly Gly Leu Pro Asp Pro
1070 1075 1080
Gly Thr Pro Ile Ala Ala Asp Leu Ala Ala Ser Ser Thr Val Met
1085 1090 1095
Trp Ser Ser Asp Thr Gly Gly Asn Ser Leu Ala Ser Gly Pro Asp
1100 1105 1110
Gly Val Lys Arg Lys Val Cys Tyr Phe Tyr Asp Pro Glu Val Gly
1115 1120 1125
Asn Tyr Tyr Tyr Gly Gln Gly His Pro Met Lys Pro His Arg Ile
1130 1135 1140
Arg Met Thr His Ala Leu Leu Ala His Tyr Gly Leu Leu Gln His
1145 1150 1155
Met Gln Val Leu Lys Pro Phe Pro Ala Arg Asp Arg Asp Leu Cys
1160 1165 1170
Arg Phe His Ala Asp Asp Tyr Val Ser Phe Leu Arg Ser Ile Thr
1175 1180 1185
Pro Glu Thr Gln Gln Asp Gln Ile Arg Gln Leu Lys Arg Phe Asn
1190 1195 1200
Val Gly Glu Asp Cys Pro Val Phe Asp Gly Leu Tyr Ser Phe Cys
1205 1210 1215
Gln Thr Tyr Ala Gly Gly Ser Val Gly Gly Ser Val Lys Leu Asn
1220 1225 1230
His Gly Leu Cys Asp Ile Ala Ile Asn Trp Ala Gly Gly Leu His
1235 1240 1245
His Ala Lys Lys Cys Glu Ala Ser Gly Phe Cys Tyr Val Asn Asp
1250 1255 1260
Ile Val Leu Ala Ile Leu Glu Leu Leu Lys Gln His Glu Arg Val
1265 1270 1275
Leu Tyr Val Asp Ile Asp Ile His His Gly Asp Gly Val Glu Glu
1280 1285 1290
Ala Phe Tyr Ala Thr Asp Arg Val Met Thr Val Ser Phe His Lys
1295 1300 1305
Phe Gly Asp Tyr Phe Pro Gly Thr Gly His Ile Gln Asp Ile Gly
1310 1315 1320
Tyr Gly Ser Gly Lys Tyr Tyr Ser Leu Asn Val Pro Leu Asp Asp
1325 1330 1335
Gly Ile Asp Asp Glu Ser Tyr His Leu Leu Phe Lys Pro Ile Met
1340 1345 1350
Gly Lys Val Met Glu Ile Phe Arg Pro Gly Ala Val Val Leu Gln
1355 1360 1365
Cys Gly Ala Asp Ser Leu Ser Gly Asp Arg Leu Gly Cys Phe Asn
1370 1375 1380
Leu Ser Ile Lys Gly His Ala Glu Cys Val Lys Phe Met Arg Ser
1385 1390 1395
Phe Asn Val Pro Leu Leu Leu Leu Gly Gly Gly Gly Tyr Thr Ile
1400 1405 1410
Arg Asn Val Ala Arg Cys Trp Cys Tyr Glu Thr Gly Val Ala Leu
1415 1420 1425
Gly Val Glu Val Glu Asp Lys Met Pro Glu His Glu Tyr Tyr Glu
1430 1435 1440
Tyr Phe Gly Pro Asp Tyr Thr Leu His Val Ala Pro Ser Asn Met
1445 1450 1455
Glu Asn Lys Asn Ser Arg Gln Met Leu Glu Glu Ile Arg Asn Asp
1460 1465 1470
Leu Leu His Asn Leu Ser Lys Leu Gln His Ala Pro Ser Val Pro
1475 1480 1485
Phe Gln Glu Arg Pro Pro Asp Thr Glu Thr Pro Glu Val Asp Glu
1490 1495 1500
Asp Gln Glu Asp Gly Asp Lys Arg Trp Asp Pro Asp Ser Asp Met
1505 1510 1515
Asp Val Asp Asp Asp Arg Lys Pro Ile Pro Ser Arg Val Lys Arg
1520 1525 1530
Glu Ala Val Glu Pro Asp Thr Lys Asp Lys Asp Gly Leu Lys Gly
1535 1540 1545
Ile Met Glu Arg Gly Lys Gly Cys Glu Val Glu Val Asp Glu Ser
1550 1555 1560
Gly Ser Thr Lys Val Lys
1565
<210> 19
<211> 2634
<212> PRT
<213> IR-vTALE-AtMET1氨基酸序列
<400> 19
Met Asp Pro Ile Arg Ser Arg Thr Pro Ser Pro Ala Arg Glu Leu Leu
1 5 10 15
Pro Gly Pro Gln Pro Asp Arg Val Gln Pro Thr Ala Asp Arg Gly Gly
20 25 30
Ala Pro Pro Ala Gly Gly Pro Leu Asp Gly Leu Pro Ala Arg Arg Thr
35 40 45
Met Ser Arg Thr Arg Leu Pro Ser Pro Pro Ala Pro Ser Pro Ala Phe
50 55 60
Ser Ala Gly Ser Phe Asn Asp Leu Leu Arg Gln Phe Asp Pro Ser Leu
65 70 75 80
Leu Asp Thr Ser Leu Leu Asp Ser Met Pro Ala Val Gly Thr Pro His
85 90 95
Thr Ala Ala Ala Pro Ala Glu Trp Asp Glu Val Gln Ser Gly Leu Arg
100 105 110
Ala Ala Asp Asp Pro Pro Pro Thr Val Arg Val Ala Val Thr Ala Ala
115 120 125
Arg Pro Pro Arg Ala Lys Pro Ala Pro Arg Arg Arg Ala Ala Gln Pro
130 135 140
Ser Asp Ala Ser Pro Ala Ala Gln Val Asp Leu Arg Thr Leu Gly Tyr
145 150 155 160
Ser Gln Gln Gln Gln Glu Lys Ile Lys Ser Lys Val Arg Ser Thr Val
165 170 175
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
180 185 190
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
195 200 205
Lys Tyr Gln His Ile Ile Thr Ala Leu Pro Glu Ala Thr His Glu Asp
210 215 220
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
225 230 235 240
Leu Leu Thr Lys Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
245 250 255
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
260 265 270
Glu Ala Val His Ala Ser Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
275 280 285
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys
290 295 300
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
305 310 315 320
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly
325 330 335
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
340 345 350
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn
355 360 365
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
370 375 380
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
385 390 395 400
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
405 410 415
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
420 425 430
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
435 440 445
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
450 455 460
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
465 470 475 480
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
485 490 495
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
500 505 510
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
515 520 525
Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
530 535 540
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
545 550 555 560
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
565 570 575
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
580 585 590
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
595 600 605
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
610 615 620
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn
625 630 635 640
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
645 650 655
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
660 665 670
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
675 680 685
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
690 695 700
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
705 710 715 720
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
725 730 735
Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val
740 745 750
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
755 760 765
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
770 775 780
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
785 790 795 800
Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
805 810 815
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
820 825 830
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
835 840 845
Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala
850 855 860
Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly
865 870 875 880
Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Pro His Ala Pro
885 890 895
Glu Leu Ile Arg Arg Ile Asn Arg Arg Ile Pro Glu Arg Thr Ser His
900 905 910
Arg Val Ala Asp Leu Ala His Val Val Arg Val Leu Gly Phe Phe Gln
915 920 925
Ser His Ser His Pro Ala Gln Ala Phe Asp Asp Ala Met Thr Gln Phe
930 935 940
Gly Met Ser Arg His Gly Leu Val Gln Leu Phe Arg Arg Val Gly Val
945 950 955 960
Thr Glu Phe Glu Ala Arg Cys Gly Thr Leu Pro Pro Ala Ser Gln Arg
965 970 975
Trp Asp Arg Ile Leu Gln Ala Ser Gly Met Lys Arg Ala Lys Pro Ser
980 985 990
Pro Thr Ser Ala Gln Thr Pro Asp Gln Ala Ser Leu His Ala Phe Ala
995 1000 1005
Asp Ser Leu Glu Arg Asp Leu Asp Ala Pro Ser Pro Met His Glu
1010 1015 1020
Gly Asp Gln Thr Arg Ala Ser Ser Arg Lys Arg Ser Arg Ser Asp
1025 1030 1035
Arg Ala Val Thr Gly Pro Ser Thr Gln Gln Ser Phe Glu Val Arg
1040 1045 1050
Val Pro Glu Gln Arg Asp Ala Leu His Leu Pro Leu Ser Trp Arg
1055 1060 1065
Val Lys Arg Pro Arg Thr Arg Ile Gly Gly Gly Leu Pro Asp Pro
1070 1075 1080
Gly Thr Pro Ile Ala Ala Asp Leu Ala Ala Ser Ser Thr Val Met
1085 1090 1095
Trp Ser Ser Val Glu Asn Gly Ala Lys Ala Ala Lys Arg Lys Lys
1100 1105 1110
Arg Pro Leu Pro Glu Ile Gln Glu Val Glu Asp Val Pro Arg Thr
1115 1120 1125
Arg Arg Pro Arg Arg Ala Ala Ala Cys Thr Ser Phe Lys Glu Lys
1130 1135 1140
Ser Ile Arg Val Cys Glu Lys Ser Ala Thr Ile Glu Val Lys Lys
1145 1150 1155
Gln Gln Ile Val Glu Glu Glu Phe Leu Ala Leu Arg Leu Thr Ala
1160 1165 1170
Leu Glu Thr Asp Val Glu Asp Arg Pro Thr Arg Arg Leu Asn Asp
1175 1180 1185
Phe Val Leu Phe Asp Ser Asp Gly Val Pro Gln Pro Leu Glu Met
1190 1195 1200
Leu Glu Ile His Asp Ile Phe Val Ser Gly Ala Ile Leu Pro Ser
1205 1210 1215
Asp Val Cys Thr Asp Lys Glu Lys Glu Lys Gly Val Arg Cys Thr
1220 1225 1230
Ser Phe Gly Arg Val Glu His Trp Ser Ile Ser Gly Tyr Glu Asp
1235 1240 1245
Gly Ser Pro Val Ile Trp Ile Ser Thr Glu Leu Ala Asp Tyr Asp
1250 1255 1260
Cys Arg Lys Pro Ala Ala Ser Tyr Arg Lys Val Tyr Asp Tyr Phe
1265 1270 1275
Tyr Glu Lys Ala Arg Ala Ser Val Ala Val Tyr Lys Lys Leu Ser
1280 1285 1290
Lys Ser Ser Gly Gly Asp Pro Asp Ile Gly Leu Glu Glu Leu Leu
1295 1300 1305
Ala Ala Val Val Arg Ser Met Ser Ser Gly Ser Lys Tyr Phe Ser
1310 1315 1320
Ser Gly Ala Ala Ile Ile Asp Phe Val Ile Ser Gln Gly Asp Phe
1325 1330 1335
Ile Tyr Asn Gln Leu Ala Gly Leu Asp Glu Thr Ala Lys Lys His
1340 1345 1350
Glu Ser Ser Tyr Val Glu Ile Pro Val Leu Val Ala Leu Arg Glu
1355 1360 1365
Lys Ser Ser Lys Ile Asp Lys Pro Leu Gln Arg Glu Arg Asn Pro
1370 1375 1380
Ser Asn Gly Val Arg Ile Lys Glu Val Ser Gln Val Ala Glu Ser
1385 1390 1395
Glu Ala Leu Thr Ser Asp Gln Leu Val Asp Gly Thr Asp Asp Asp
1400 1405 1410
Arg Arg Tyr Ala Ile Leu Leu Gln Asp Glu Glu Asn Arg Lys Ser
1415 1420 1425
Met Gln Gln Pro Arg Lys Asn Ser Ser Ser Gly Ser Ala Ser Asn
1430 1435 1440
Met Phe Tyr Ile Lys Ile Asn Glu Asp Glu Ile Ala Asn Asp Tyr
1445 1450 1455
Pro Leu Pro Ser Tyr Tyr Lys Thr Ser Glu Glu Glu Thr Asp Glu
1460 1465 1470
Leu Ile Leu Tyr Asp Ala Ser Tyr Glu Val Gln Ser Glu His Leu
1475 1480 1485
Pro His Arg Met Leu His Asn Trp Ala Leu Tyr Asn Ser Asp Leu
1490 1495 1500
Arg Phe Ile Ser Leu Glu Leu Leu Pro Met Lys Gln Cys Asp Asp
1505 1510 1515
Ile Asp Val Asn Ile Phe Gly Ser Gly Val Val Thr Asp Asp Asn
1520 1525 1530
Gly Ser Trp Ile Ser Leu Asn Asp Pro Asp Ser Gly Ser Gln Ser
1535 1540 1545
His Asp Pro Asp Gly Met Cys Ile Phe Leu Ser Gln Ile Lys Glu
1550 1555 1560
Trp Met Ile Glu Phe Gly Ser Asp Asp Ile Ile Ser Ile Ser Ile
1565 1570 1575
Arg Thr Asp Val Ala Trp Tyr Arg Leu Gly Lys Pro Ser Lys Leu
1580 1585 1590
Tyr Ala Pro Trp Trp Lys Pro Val Leu Lys Thr Ala Arg Val Gly
1595 1600 1605
Ile Ser Ile Leu Thr Phe Leu Arg Val Glu Ser Arg Val Ala Arg
1610 1615 1620
Leu Ser Phe Ala Asp Val Thr Lys Arg Leu Ser Gly Leu Gln Ala
1625 1630 1635
Asn Asp Lys Ala Tyr Ile Ser Ser Asp Pro Leu Ala Val Glu Arg
1640 1645 1650
Tyr Leu Val Val His Gly Gln Ile Ile Leu Gln Leu Phe Ala Val
1655 1660 1665
Tyr Pro Asp Asp Asn Val Lys Arg Cys Pro Phe Val Val Gly Leu
1670 1675 1680
Ala Ser Lys Leu Glu Asp Arg His His Thr Lys Trp Ile Ile Lys
1685 1690 1695
Lys Lys Lys Ile Ser Leu Lys Glu Leu Asn Leu Asn Pro Arg Ala
1700 1705 1710
Gly Met Ala Pro Val Ala Ser Lys Arg Lys Ala Met Gln Ala Thr
1715 1720 1725
Thr Thr Arg Leu Val Asn Arg Ile Trp Gly Glu Phe Tyr Ser Asn
1730 1735 1740
Tyr Ser Pro Glu Asp Pro Leu Gln Ala Thr Ala Ala Glu Asn Gly
1745 1750 1755
Glu Asp Glu Val Glu Glu Glu Gly Gly Asn Gly Glu Glu Glu Val
1760 1765 1770
Glu Glu Glu Gly Glu Asn Gly Leu Thr Glu Asp Thr Val Pro Glu
1775 1780 1785
Pro Val Glu Val Gln Lys Pro His Thr Pro Lys Lys Ile Arg Gly
1790 1795 1800
Ser Ser Gly Lys Arg Glu Ile Lys Trp Asp Gly Glu Ser Leu Gly
1805 1810 1815
Lys Thr Ser Ala Gly Glu Pro Leu Tyr Gln Gln Ala Leu Val Gly
1820 1825 1830
Gly Glu Met Val Ala Val Gly Gly Ala Val Thr Leu Glu Val Asp
1835 1840 1845
Asp Pro Asp Glu Met Pro Ala Ile Tyr Phe Val Glu Tyr Met Phe
1850 1855 1860
Glu Ser Thr Asp His Cys Lys Met Leu His Gly Arg Phe Leu Gln
1865 1870 1875
Arg Gly Ser Met Thr Val Leu Gly Asn Ala Ala Asn Glu Arg Glu
1880 1885 1890
Leu Phe Leu Thr Asn Glu Cys Met Thr Thr Gln Leu Lys Asp Ile
1895 1900 1905
Lys Gly Val Ala Ser Phe Glu Ile Arg Ser Arg Pro Trp Gly His
1910 1915 1920
Gln Tyr Arg Lys Lys Asn Ile Thr Ala Asp Lys Leu Asp Trp Ala
1925 1930 1935
Arg Ala Leu Glu Arg Lys Val Lys Asp Leu Pro Thr Glu Tyr Tyr
1940 1945 1950
Cys Lys Ser Leu Tyr Ser Pro Glu Arg Gly Gly Phe Phe Ser Leu
1955 1960 1965
Pro Leu Ser Asp Ile Gly Arg Ser Ser Gly Phe Cys Thr Ser Cys
1970 1975 1980
Lys Ile Arg Glu Asp Glu Glu Lys Arg Ser Thr Ile Lys Leu Asn
1985 1990 1995
Val Ser Lys Thr Gly Phe Phe Ile Asn Gly Ile Glu Tyr Ser Val
2000 2005 2010
Glu Asp Phe Val Tyr Val Asn Pro Asp Ser Ile Gly Gly Leu Lys
2015 2020 2025
Glu Gly Ser Lys Thr Ser Phe Lys Ser Gly Arg Asn Ile Gly Leu
2030 2035 2040
Arg Ala Tyr Val Val Cys Gln Leu Leu Glu Ile Val Pro Lys Glu
2045 2050 2055
Ser Arg Lys Ala Asp Leu Gly Ser Phe Asp Val Lys Val Arg Arg
2060 2065 2070
Phe Tyr Arg Pro Glu Asp Val Ser Ala Glu Lys Ala Tyr Ala Ser
2075 2080 2085
Asp Ile Gln Glu Leu Tyr Phe Ser Gln Asp Thr Val Val Leu Pro
2090 2095 2100
Pro Gly Ala Leu Glu Gly Lys Cys Glu Val Arg Lys Lys Ser Asp
2105 2110 2115
Met Pro Leu Ser Arg Glu Tyr Pro Ile Ser Asp His Ile Phe Phe
2120 2125 2130
Cys Asp Leu Phe Phe Asp Thr Ser Lys Gly Ser Leu Lys Gln Leu
2135 2140 2145
Pro Ala Asn Met Lys Pro Lys Phe Ser Thr Ile Lys Asp Asp Thr
2150 2155 2160
Leu Leu Arg Lys Lys Lys Gly Lys Gly Val Glu Ser Glu Ile Glu
2165 2170 2175
Ser Glu Ile Val Lys Pro Val Glu Pro Pro Lys Glu Ile Arg Leu
2180 2185 2190
Ala Thr Leu Asp Ile Phe Ala Gly Cys Gly Gly Leu Ser His Gly
2195 2200 2205
Leu Lys Lys Ala Gly Val Ser Asp Ala Lys Trp Ala Ile Glu Tyr
2210 2215 2220
Glu Glu Pro Ala Gly Gln Ala Phe Lys Gln Asn His Pro Glu Ser
2225 2230 2235
Thr Val Phe Val Asp Asn Cys Asn Val Ile Leu Arg Ala Ile Met
2240 2245 2250
Glu Lys Gly Gly Asp Gln Asp Asp Cys Val Ser Thr Thr Glu Ala
2255 2260 2265
Asn Glu Leu Ala Ala Lys Leu Thr Glu Glu Gln Lys Ser Thr Leu
2270 2275 2280
Pro Leu Pro Gly Gln Val Asp Phe Ile Asn Gly Gly Pro Pro Cys
2285 2290 2295
Gln Gly Phe Ser Gly Met Asn Arg Phe Asn Gln Ser Ser Trp Ser
2300 2305 2310
Lys Val Gln Cys Glu Met Ile Leu Ala Phe Leu Ser Phe Ala Asp
2315 2320 2325
Tyr Phe Arg Pro Arg Tyr Phe Leu Leu Glu Asn Val Arg Thr Phe
2330 2335 2340
Val Ser Phe Asn Lys Gly Gln Thr Phe Gln Leu Thr Leu Ala Ser
2345 2350 2355
Leu Leu Glu Met Gly Tyr Gln Val Arg Phe Gly Ile Leu Glu Ala
2360 2365 2370
Gly Ala Tyr Gly Val Ser Gln Ser Arg Lys Arg Ala Phe Ile Trp
2375 2380 2385
Ala Ala Ala Pro Glu Glu Val Leu Pro Glu Trp Pro Glu Pro Met
2390 2395 2400
His Val Phe Gly Val Pro Lys Leu Lys Ile Ser Leu Ser Gln Gly
2405 2410 2415
Leu His Tyr Ala Ala Val Arg Ser Thr Ala Leu Gly Ala Pro Phe
2420 2425 2430
Arg Pro Ile Thr Val Arg Asp Thr Ile Gly Asp Leu Pro Ser Val
2435 2440 2445
Glu Asn Gly Asp Ser Arg Thr Asn Lys Glu Tyr Lys Glu Val Ala
2450 2455 2460
Val Ser Trp Phe Gln Lys Glu Ile Arg Gly Asn Thr Ile Ala Leu
2465 2470 2475
Thr Asp His Ile Cys Lys Ala Met Asn Glu Leu Asn Leu Ile Arg
2480 2485 2490
Cys Lys Leu Ile Pro Thr Arg Pro Gly Ala Asp Trp His Asp Leu
2495 2500 2505
Pro Lys Arg Lys Val Thr Leu Ser Asp Gly Arg Val Glu Glu Met
2510 2515 2520
Ile Pro Phe Cys Leu Pro Asn Thr Ala Glu Arg His Asn Gly Trp
2525 2530 2535
Lys Gly Leu Tyr Gly Arg Leu Asp Trp Gln Gly Asn Phe Pro Thr
2540 2545 2550
Ser Val Thr Asp Pro Gln Pro Met Gly Lys Val Gly Met Cys Phe
2555 2560 2565
His Pro Glu Gln His Arg Ile Leu Thr Val Arg Glu Cys Ala Arg
2570 2575 2580
Ser Gln Gly Phe Pro Asp Ser Tyr Glu Phe Ala Gly Asn Ile Asn
2585 2590 2595
His Lys His Arg Gln Ile Gly Asn Ala Val Pro Pro Pro Leu Ala
2600 2605 2610
Phe Ala Leu Gly Arg Lys Leu Lys Glu Ala Leu His Leu Lys Lys
2615 2620 2625
Ser Pro Gln His Gln Pro
2630
<210> 20
<211> 3330
<212> DNA
<213> RB-vTALE-MCS核苷酸序列
<400> 20
atggacccca ttcgttcgcg cacgccaagt cctgcccgcg agcttctgcc cggaccccaa 60
ccggataggg ttcagccgac tgcagatcgg gggggggctc cgcctgctgg cggccccctg 120
gatggcttgc ccgctcggcg gacgatgtcc cggacccggc tgccatctcc ccctgcgccc 180
tcgcctgcgt tctcggcggg cagcttcaac gatctgctcc gtcagttcga tccgtcgctt 240
cttgatacat cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc 300
ccagcagagt gggatgaggt gcaatcgggt ctgcgtgcag ccgatgaccc gccacccacc 360
gtgcgtgtcg ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt 420
gcggcgcaac cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac 480
agtcagcagc agcaagagaa gatcaaatcg aaggtgcgtt cgacagtggc gcagcaccac 540
gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg 600
gcagcgttag ggaccgtcgc tgtcaagtat cagcacataa tcacggcgtt gccagaggcg 660
acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc 720
ttgctcacga aggcggggga gttgagaggt ccgccgttac agttggacac aggccaactt 780
ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc atcgcgcaat 840
gcactgacgg gtgcccccct gaaccttact ccagatcaag ttgtggcaat cgcttcaaat 900
atcggaggga aacaggcctt ggaaaccgtg caacgtcttc ttcctgtgtt gtgtcaagct 960
catggactca ctccagatca agttgtggct atcgctagta acggtggtgg aaaacaggcg 1020
ttagaaactg tccaaaggct ccttcccgtg ttgtgtcaag ctcatggcct cactccagat 1080
caagttgtgg caatcgcttc aaatatcgga gggaaacagg ccttggaaac cgtgcaacgt 1140
cttcttcctg tgttgtgtca agctcatgga ttaactccag atcaagttgt ggctatcgct 1200
agtaacggtg gtggaaaaca ggcgttagaa actgtccaaa ggctccttcc cgtgttgtgt 1260
caagctcatg gcttaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 1320
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggactcact 1380
ccagatcaag ttgtggctat cgctagtaac ggtggtggaa aacaggcgtt agaaactgtc 1440
caaaggctcc ttcccgtgtt gtgtcaagct catggcctca ctccagatca agttgtggct 1500
atcgctagta acggtggtgg aaaacaggcg ttagaaactg tccaaaggct ccttcccgtg 1560
ttgtgtcaag ctcatggact cactccagat caagttgtgg caatagcctc taacaatgga 1620
ggtaaacaag cacttgagac tgttcagaga ttgttaccgg tgttgtgtca agctcatggc 1680
ctcactccag atcaagttgt ggcaatagcc tctaacaatg gaggtaaaca agcacttgag 1740
actgttcaga gattgttacc ggtgttgtgt caagctcatg gattaactcc agatcaagtt 1800
gtggcaatcg cttcaaatat cggagggaaa caggccttgg aaaccgtgca acgtcttctt 1860
cctgtgttgt gtcaagctca tggtctcact ccagatcaag ttgtggccat tgcatctcat 1920
gatggtggta agcaagctct cgaaacagtc caaagacttc tcccagtgtt gtgtcaagct 1980
catggattaa ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 2040
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggtct cactccagat 2100
caagttgtgg ccattgcatc tcatgatggt ggtaagcaag ctctcgaaac agtccaaaga 2160
cttctcccag tgttgtgtca agctcatgga ctcactccag atcaagttgt ggccattgca 2220
tctcatgatg gtggtaagca agctctcgaa acagtccaaa gacttctccc agtgttgtgt 2280
caagctcatg gattaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 2340
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggtttaact 2400
ccagatcaag ttgtggcaat cgcttcaaat atcggaggga aacaggcctt ggaaaccgtg 2460
caacgtcttc ttcctgtgtt gtgtcaagct catggtctca ctccagatca agttgtggca 2520
atagcctcta acaatggagg taaacaagca cttgagagca ttgttgccca gttatctcgc 2580
cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc 2640
ggacgtcctg ccctggatgc agtgaaaaag ggattgccgc acgcgccgga attgatcaga 2700
agaatcaatc gccgtattcc cgaacgcacg tcccatcgcg ttgccgacct cgcgcacgtg 2760
gtgcgcgtgc ttggtttttt ccagagccac tcccacccag cgcaagcatt cgatgacgcc 2820
atgacgcagt tcgggatgag caggcacggg ttggtacagc tctttcgcag agtgggcgtc 2880
accgaattcg aagcccgctg cggaacgctc cccccagcct cgcagcgttg ggaccgtatc 2940
ctccaggcat cagggatgaa aagggccaaa ccgtccccta cttcagctca aacgccggat 3000
caggcgtctt tgcatgcatt cgccgattcg ctggagcgtg accttgatgc gcccagccca 3060
atgcacgagg gagatcagac gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct 3120
gtcaccggcc cctccacaca gcaatctttc gaggtgcgcg ttcccgaaca gcgcgatgcg 3180
ctgcatttgc ccctcagctg gagggtaaaa cgcccgcgta ccaggatcgg gggcggcctc 3240
ccggatcctg gtacgcccat cgctgccgac ctggcagcgt ccagcaccgt gatgtggtct 3300
agattaatta accccggggt cgactagtga 3330
<210> 21
<211> 5175
<212> DNA
<213> RB-vTALE-AtKYP核苷酸序列
<400> 21
atggacccca ttcgttcgcg cacgccaagt cctgcccgcg agcttctgcc cggaccccaa 60
ccggataggg ttcagccgac tgcagatcgg gggggggctc cgcctgctgg cggccccctg 120
gatggcttgc ccgctcggcg gacgatgtcc cggacccggc tgccatctcc ccctgcgccc 180
tcgcctgcgt tctcggcggg cagcttcaac gatctgctcc gtcagttcga tccgtcgctt 240
cttgatacat cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc 300
ccagcagagt gggatgaggt gcaatcgggt ctgcgtgcag ccgatgaccc gccacccacc 360
gtgcgtgtcg ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt 420
gcggcgcaac cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac 480
agtcagcagc agcaagagaa gatcaaatcg aaggtgcgtt cgacagtggc gcagcaccac 540
gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg 600
gcagcgttag ggaccgtcgc tgtcaagtat cagcacataa tcacggcgtt gccagaggcg 660
acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc 720
ttgctcacga aggcggggga gttgagaggt ccgccgttac agttggacac aggccaactt 780
ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc atcgcgcaat 840
gcactgacgg gtgcccccct gaaccttact ccagatcaag ttgtggcaat cgcttcaaat 900
atcggaggga aacaggcctt ggaaaccgtg caacgtcttc ttcctgtgtt gtgtcaagct 960
catggactca ctccagatca agttgtggct atcgctagta acggtggtgg aaaacaggcg 1020
ttagaaactg tccaaaggct ccttcccgtg ttgtgtcaag ctcatggcct cactccagat 1080
caagttgtgg caatcgcttc aaatatcgga gggaaacagg ccttggaaac cgtgcaacgt 1140
cttcttcctg tgttgtgtca agctcatgga ttaactccag atcaagttgt ggctatcgct 1200
agtaacggtg gtggaaaaca ggcgttagaa actgtccaaa ggctccttcc cgtgttgtgt 1260
caagctcatg gcttaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 1320
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggactcact 1380
ccagatcaag ttgtggctat cgctagtaac ggtggtggaa aacaggcgtt agaaactgtc 1440
caaaggctcc ttcccgtgtt gtgtcaagct catggcctca ctccagatca agttgtggct 1500
atcgctagta acggtggtgg aaaacaggcg ttagaaactg tccaaaggct ccttcccgtg 1560
ttgtgtcaag ctcatggact cactccagat caagttgtgg caatagcctc taacaatgga 1620
ggtaaacaag cacttgagac tgttcagaga ttgttaccgg tgttgtgtca agctcatggc 1680
ctcactccag atcaagttgt ggcaatagcc tctaacaatg gaggtaaaca agcacttgag 1740
actgttcaga gattgttacc ggtgttgtgt caagctcatg gattaactcc agatcaagtt 1800
gtggcaatcg cttcaaatat cggagggaaa caggccttgg aaaccgtgca acgtcttctt 1860
cctgtgttgt gtcaagctca tggtctcact ccagatcaag ttgtggccat tgcatctcat 1920
gatggtggta agcaagctct cgaaacagtc caaagacttc tcccagtgtt gtgtcaagct 1980
catggattaa ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 2040
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggtct cactccagat 2100
caagttgtgg ccattgcatc tcatgatggt ggtaagcaag ctctcgaaac agtccaaaga 2160
cttctcccag tgttgtgtca agctcatgga ctcactccag atcaagttgt ggccattgca 2220
tctcatgatg gtggtaagca agctctcgaa acagtccaaa gacttctccc agtgttgtgt 2280
caagctcatg gattaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 2340
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggtttaact 2400
ccagatcaag ttgtggcaat cgcttcaaat atcggaggga aacaggcctt ggaaaccgtg 2460
caacgtcttc ttcctgtgtt gtgtcaagct catggtctca ctccagatca agttgtggca 2520
atagcctcta acaatggagg taaacaagca cttgagagca ttgttgccca gttatctcgc 2580
cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc 2640
ggacgtcctg ccctggatgc agtgaaaaag ggattgccgc acgcgccgga attgatcaga 2700
agaatcaatc gccgtattcc cgaacgcacg tcccatcgcg ttgccgacct cgcgcacgtg 2760
gtgcgcgtgc ttggtttttt ccagagccac tcccacccag cgcaagcatt cgatgacgcc 2820
atgacgcagt tcgggatgag caggcacggg ttggtacagc tctttcgcag agtgggcgtc 2880
accgaattcg aagcccgctg cggaacgctc cccccagcct cgcagcgttg ggaccgtatc 2940
ctccaggcat cagggatgaa aagggccaaa ccgtccccta cttcagctca aacgccggat 3000
caggcgtctt tgcatgcatt cgccgattcg ctggagcgtg accttgatgc gcccagccca 3060
atgcacgagg gagatcagac gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct 3120
gtcaccggcc cctccacaca gcaatctttc gaggtgcgcg ttcccgaaca gcgcgatgcg 3180
ctgcatttgc ccctcagctg gagggtaaaa cgcccgcgta ccaggatcgg gggcggcctc 3240
ccggatcctg gtacgcccat cgctgccgac ctggcagcgt ccagcaccgt gatgtggtct 3300
agtgctggaa aaaggaaacg agctaatgct cctgaccaaa cagagcgaag atcgagtgtt 3360
cgggttcaga aagtgagaca gaaagcgtta gatgagaagg cgcgtttagt acaggagagg 3420
gttaagctcc tcagtgacag aaagagtgaa atttgtgtcg atgacactga gttacatgag 3480
aaagaagagg aaaatgtcga tgggagccct aaacgaagaa gccctccaaa gctaaccgca 3540
atgcagaaag gaaagcagaa attgagtgtt tctctgaatg gtaaggacgt gaacttggaa 3600
cctcatctca aagtgacaaa gtgtctgagg ttatttaaca agcaatatct cctctgtgtc 3660
caggctaagt tgagcaggcc tgatttgaag ggtgtaactg agatgataaa agctaaggcg 3720
atattgtacc caagaaaaat aatcggtgac cttccaggta tagacgttgg acaccgtttt 3780
ttttcaagag ctgaaatgtg tgctgtagga ttccataacc attggctaaa tggcattgat 3840
tatatgtcaa tggaatacga aaaagagtat agtaactaca aattaccgct tgctgtttct 3900
attgttatgt cgggccagta cgaggatgat ctagacaatg cagatacagt gacttacact 3960
ggacagggag ggcataactt aactggtaat aaacgtcaga taaaggatca acttttagaa 4020
cgagggaatt tggcgctaaa gcactgctgc gaatataatg tgcctgtcag agtaactcgt 4080
ggtcacaatt gcaaaagtag ctataccaaa cgagtataca cttatgatgg actgtacaag 4140
gttgaaaagt tctgggcaca aaagggcgtt tcaggattta cagtgtataa gtaccgactg 4200
aaacgattgg aggggcaacc agaactaact actgatcagg tcaactttgt tgctggacgc 4260
ataccaacga gtacttcaga aattgagggt ttggtatgtg aggacatctc cggagggcta 4320
gaatttaagg gtatccccgc cactaatcgt gttgatgatt caccagtttc accaacatct 4380
ggtttcacat acatcaaatc tttgattatt gagcctaatg tcataattcc aaagagttca 4440
actgggtgta actgccgagg cagctgcact gactcaaaga aatgtgcatg tgctaagctt 4500
aatgggggta actttccata tgttgacctt aatgatggca gattaattga gtctcgagat 4560
gttgtatttg aatgtggtcc tcactgtggg tgtgggccaa aatgtgtcaa ccgaacttct 4620
cagaagcgtc taagattcaa tcttgaggtt ttccgctctg caaagaaggg ttgggcagtt 4680
agatcatggg agtacatacc agctggttca ccagtatgtg agtacatagg agttgtcagg 4740
agaactgctg atgtggatac tatctctgac aatgaataca tatttgagat tgactgccaa 4800
cagacaatgc aaggtcttgg tggaagacag agaagactaa gagatgttgc tgtaccaatg 4860
aataatggag tcagtcagag cagtgaagat gagaatgcgc cagagttctg cattgatgct 4920
ggttcaacag gaaactttgc taggtttata aatcacagtt gtgaaccaaa cctatttgtt 4980
cagtgcgtcc tgagttctca ccaggatata aggcttgccc gtgtggttct tttcgcagct 5040
gacaacattt ccccaatgca ggagctcact tacgactatg gatatgcgct tgatagcgtt 5100
catggaccgg atgggaaggt gaagcagctc gcttgctact gtggagcgct aaattgtagg 5160
aaacgccttt actaa 5175
<210> 22
<211> 4710
<212> DNA
<213> RB-vTALE-AtHDAC19核苷酸序列
<400> 22
atggacccca ttcgttcgcg cacgccaagt cctgcccgcg agcttctgcc cggaccccaa 60
ccggataggg ttcagccgac tgcagatcgg gggggggctc cgcctgctgg cggccccctg 120
gatggcttgc ccgctcggcg gacgatgtcc cggacccggc tgccatctcc ccctgcgccc 180
tcgcctgcgt tctcggcggg cagcttcaac gatctgctcc gtcagttcga tccgtcgctt 240
cttgatacat cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc 300
ccagcagagt gggatgaggt gcaatcgggt ctgcgtgcag ccgatgaccc gccacccacc 360
gtgcgtgtcg ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt 420
gcggcgcaac cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac 480
agtcagcagc agcaagagaa gatcaaatcg aaggtgcgtt cgacagtggc gcagcaccac 540
gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg 600
gcagcgttag ggaccgtcgc tgtcaagtat cagcacataa tcacggcgtt gccagaggcg 660
acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc 720
ttgctcacga aggcggggga gttgagaggt ccgccgttac agttggacac aggccaactt 780
ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc atcgcgcaat 840
gcactgacgg gtgcccccct gaaccttact ccagatcaag ttgtggcaat cgcttcaaat 900
atcggaggga aacaggcctt ggaaaccgtg caacgtcttc ttcctgtgtt gtgtcaagct 960
catggactca ctccagatca agttgtggct atcgctagta acggtggtgg aaaacaggcg 1020
ttagaaactg tccaaaggct ccttcccgtg ttgtgtcaag ctcatggcct cactccagat 1080
caagttgtgg caatcgcttc aaatatcgga gggaaacagg ccttggaaac cgtgcaacgt 1140
cttcttcctg tgttgtgtca agctcatgga ttaactccag atcaagttgt ggctatcgct 1200
agtaacggtg gtggaaaaca ggcgttagaa actgtccaaa ggctccttcc cgtgttgtgt 1260
caagctcatg gcttaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 1320
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggactcact 1380
ccagatcaag ttgtggctat cgctagtaac ggtggtggaa aacaggcgtt agaaactgtc 1440
caaaggctcc ttcccgtgtt gtgtcaagct catggcctca ctccagatca agttgtggct 1500
atcgctagta acggtggtgg aaaacaggcg ttagaaactg tccaaaggct ccttcccgtg 1560
ttgtgtcaag ctcatggact cactccagat caagttgtgg caatagcctc taacaatgga 1620
ggtaaacaag cacttgagac tgttcagaga ttgttaccgg tgttgtgtca agctcatggc 1680
ctcactccag atcaagttgt ggcaatagcc tctaacaatg gaggtaaaca agcacttgag 1740
actgttcaga gattgttacc ggtgttgtgt caagctcatg gattaactcc agatcaagtt 1800
gtggcaatcg cttcaaatat cggagggaaa caggccttgg aaaccgtgca acgtcttctt 1860
cctgtgttgt gtcaagctca tggtctcact ccagatcaag ttgtggccat tgcatctcat 1920
gatggtggta agcaagctct cgaaacagtc caaagacttc tcccagtgtt gtgtcaagct 1980
catggattaa ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 2040
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggtct cactccagat 2100
caagttgtgg ccattgcatc tcatgatggt ggtaagcaag ctctcgaaac agtccaaaga 2160
cttctcccag tgttgtgtca agctcatgga ctcactccag atcaagttgt ggccattgca 2220
tctcatgatg gtggtaagca agctctcgaa acagtccaaa gacttctccc agtgttgtgt 2280
caagctcatg gattaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 2340
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggtttaact 2400
ccagatcaag ttgtggcaat cgcttcaaat atcggaggga aacaggcctt ggaaaccgtg 2460
caacgtcttc ttcctgtgtt gtgtcaagct catggtctca ctccagatca agttgtggca 2520
atagcctcta acaatggagg taaacaagca cttgagagca ttgttgccca gttatctcgc 2580
cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc 2640
ggacgtcctg ccctggatgc agtgaaaaag ggattgccgc acgcgccgga attgatcaga 2700
agaatcaatc gccgtattcc cgaacgcacg tcccatcgcg ttgccgacct cgcgcacgtg 2760
gtgcgcgtgc ttggtttttt ccagagccac tcccacccag cgcaagcatt cgatgacgcc 2820
atgacgcagt tcgggatgag caggcacggg ttggtacagc tctttcgcag agtgggcgtc 2880
accgaattcg aagcccgctg cggaacgctc cccccagcct cgcagcgttg ggaccgtatc 2940
ctccaggcat cagggatgaa aagggccaaa ccgtccccta cttcagctca aacgccggat 3000
caggcgtctt tgcatgcatt cgccgattcg ctggagcgtg accttgatgc gcccagccca 3060
atgcacgagg gagatcagac gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct 3120
gtcaccggcc cctccacaca gcaatctttc gaggtgcgcg ttcccgaaca gcgcgatgcg 3180
ctgcatttgc ccctcagctg gagggtaaaa cgcccgcgta ccaggatcgg gggcggcctc 3240
ccggatcctg gtacgcccat cgctgccgac ctggcagcgt ccagcaccgt gatgtggtct 3300
agtgatactg gcggcaattc gctggcgtcc ggacctgatg gtgtgaagag gaaagtttgt 3360
tatttctatg accctgaggt cggcaattac tactatggcc aaggtcatcc catgaagccc 3420
catcgcatcc gcatgaccca tgccctcctc gctcactacg gtctccttca gcatatgcag 3480
gttctcaagc ccttccctgc ccgcgaccgt gatctctgcc gcttccacgc cgacgactat 3540
gtctcttttc tccgcagcat tacccctgaa acccagcaag atcagattcg ccaacttaag 3600
cgcttcaatg ttggtgaaga ctgtcccgtc tttgacggcc tttattcctt ttgccagacc 3660
tatgctggag gatctgttgg tggctctgtc aagcttaacc acggcctctg cgatattgcc 3720
atcaactggg ctggtggtct ccatcacgct aagaagtgcg aggcctctgg cttctgttac 3780
gtcaatgata tcgtcttagc tatcctagag ctccttaagc agcatgagcg tgttctttat 3840
gtcgatattg atatccacca cggggatgga gtggaggagg cattttatgc tactgacagg 3900
gttatgactg tctcgtttca taaatttggt gattactttc ccggtacagg tcacattcag 3960
gatataggtt atggtagcgg aaagtactat tctctcaatg taccactgga tgatggaatc 4020
gatgatgaga gctatcatct gttattcaag cccatcatgg ggaaagttat ggaaattttc 4080
cgaccagggg ctgtggtatt gcaatgtggt gctgattcat tgtctggtga taggttgggg 4140
tgctttaatc tttcaatcaa aggtcatgct gagtgcgtca aatttatgag atcgttcaat 4200
gttcccctac tgctcttggg tggtggtggt tacactatcc gcaatgttgc ccgttgctgg 4260
tgctacgaga ctggagttgc acttggagtt gaagttgaag acaagatgcc ggagcatgaa 4320
tattatgaat actttggtcc agactataca cttcacgttg ctccaagtaa catggaaaat 4380
aagaattctc gtcagatgct tgaagagatt cgcaatgacc ttctccacaa tctctctaag 4440
cttcagcatg ctccaagtgt accatttcag gaaagaccac ctgatacaga gactcccgag 4500
gttgatgaag accaagaaga tggggataaa agatgggatc cggattcaga catggatgtt 4560
gatgatgacc gtaaacctat accaagcaga gtaaaaagag aagctgttga accagataca 4620
aaggacaagg atggactgaa aggaattatg gagcgtggaa aaggttgtga ggtggaggtg 4680
gatgagagtg gaagcactaa ggtaaaatga 4710
<210> 23
<211> 7905
<212> DNA
<213> RB-vTALE-AtMET1核苷酸序列
<400> 23
atggacccca ttcgttcgcg cacgccaagt cctgcccgcg agcttctgcc cggaccccaa 60
ccggataggg ttcagccgac tgcagatcgg gggggggctc cgcctgctgg cggccccctg 120
gatggcttgc ccgctcggcg gacgatgtcc cggacccggc tgccatctcc ccctgcgccc 180
tcgcctgcgt tctcggcggg cagcttcaac gatctgctcc gtcagttcga tccgtcgctt 240
cttgatacat cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc 300
ccagcagagt gggatgaggt gcaatcgggt ctgcgtgcag ccgatgaccc gccacccacc 360
gtgcgtgtcg ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt 420
gcggcgcaac cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac 480
agtcagcagc agcaagagaa gatcaaatcg aaggtgcgtt cgacagtggc gcagcaccac 540
gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg 600
gcagcgttag ggaccgtcgc tgtcaagtat cagcacataa tcacggcgtt gccagaggcg 660
acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc 720
ttgctcacga aggcggggga gttgagaggt ccgccgttac agttggacac aggccaactt 780
ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc atcgcgcaat 840
gcactgacgg gtgcccccct gaaccttact ccagatcaag ttgtggcaat cgcttcaaat 900
atcggaggga aacaggcctt ggaaaccgtg caacgtcttc ttcctgtgtt gtgtcaagct 960
catggactca ctccagatca agttgtggct atcgctagta acggtggtgg aaaacaggcg 1020
ttagaaactg tccaaaggct ccttcccgtg ttgtgtcaag ctcatggcct cactccagat 1080
caagttgtgg caatcgcttc aaatatcgga gggaaacagg ccttggaaac cgtgcaacgt 1140
cttcttcctg tgttgtgtca agctcatgga ttaactccag atcaagttgt ggctatcgct 1200
agtaacggtg gtggaaaaca ggcgttagaa actgtccaaa ggctccttcc cgtgttgtgt 1260
caagctcatg gcttaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 1320
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggactcact 1380
ccagatcaag ttgtggctat cgctagtaac ggtggtggaa aacaggcgtt agaaactgtc 1440
caaaggctcc ttcccgtgtt gtgtcaagct catggcctca ctccagatca agttgtggct 1500
atcgctagta acggtggtgg aaaacaggcg ttagaaactg tccaaaggct ccttcccgtg 1560
ttgtgtcaag ctcatggact cactccagat caagttgtgg caatagcctc taacaatgga 1620
ggtaaacaag cacttgagac tgttcagaga ttgttaccgg tgttgtgtca agctcatggc 1680
ctcactccag atcaagttgt ggcaatagcc tctaacaatg gaggtaaaca agcacttgag 1740
actgttcaga gattgttacc ggtgttgtgt caagctcatg gattaactcc agatcaagtt 1800
gtggcaatcg cttcaaatat cggagggaaa caggccttgg aaaccgtgca acgtcttctt 1860
cctgtgttgt gtcaagctca tggtctcact ccagatcaag ttgtggccat tgcatctcat 1920
gatggtggta agcaagctct cgaaacagtc caaagacttc tcccagtgtt gtgtcaagct 1980
catggattaa ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 2040
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggtct cactccagat 2100
caagttgtgg ccattgcatc tcatgatggt ggtaagcaag ctctcgaaac agtccaaaga 2160
cttctcccag tgttgtgtca agctcatgga ctcactccag atcaagttgt ggccattgca 2220
tctcatgatg gtggtaagca agctctcgaa acagtccaaa gacttctccc agtgttgtgt 2280
caagctcatg gattaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 2340
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggtttaact 2400
ccagatcaag ttgtggcaat cgcttcaaat atcggaggga aacaggcctt ggaaaccgtg 2460
caacgtcttc ttcctgtgtt gtgtcaagct catggtctca ctccagatca agttgtggca 2520
atagcctcta acaatggagg taaacaagca cttgagagca ttgttgccca gttatctcgc 2580
cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc 2640
ggacgtcctg ccctggatgc agtgaaaaag ggattgccgc acgcgccgga attgatcaga 2700
agaatcaatc gccgtattcc cgaacgcacg tcccatcgcg ttgccgacct cgcgcacgtg 2760
gtgcgcgtgc ttggtttttt ccagagccac tcccacccag cgcaagcatt cgatgacgcc 2820
atgacgcagt tcgggatgag caggcacggg ttggtacagc tctttcgcag agtgggcgtc 2880
accgaattcg aagcccgctg cggaacgctc cccccagcct cgcagcgttg ggaccgtatc 2940
ctccaggcat cagggatgaa aagggccaaa ccgtccccta cttcagctca aacgccggat 3000
caggcgtctt tgcatgcatt cgccgattcg ctggagcgtg accttgatgc gcccagccca 3060
atgcacgagg gagatcagac gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct 3120
gtcaccggcc cctccacaca gcaatctttc gaggtgcgcg ttcccgaaca gcgcgatgcg 3180
ctgcatttgc ccctcagctg gagggtaaaa cgcccgcgta ccaggatcgg gggcggcctc 3240
ccggatcctg gtacgcccat cgctgccgac ctggcagcgt ccagcaccgt gatgtggtct 3300
agtgtggaaa atggggctaa agctgcgaag cgaaagaaga gaccacttcc agagattcaa 3360
gaggtagaag atgtacctag gacgaggaga ccaaggcgtg ctgcagcgtg taccagtttc 3420
aaggagaaat ctattcgagt ctgtgagaaa tctgctacta ttgaagtaaa gaaacagcag 3480
attgtggagg aagagtttct cgcgttacgg ttaacggctc tggaaactga tgttgaagat 3540
cgtccaacca ggagactgaa tgattttgtt ttgtttgatt cagatggagt tccacaacct 3600
ctggagatgt tggagattca tgacatattc gtttcaggtg ctatcttacc ttcagatgtg 3660
tgtactgata aggagaaaga gaagggtgtg aggtgtacat cgtttggacg ggttgagcat 3720
tggagtatct ctggttatga agatggttcc cctgttattt ggatctcaac ggaattggcg 3780
gattatgatt gtcgtaaacc tgctgctagc tacaggaagg tttatgatta cttctatgag 3840
aaagctcgtg cttcagtggc tgtgtataag aaattgtcca agtcatctgg tggggatcct 3900
gatataggtc ttgaggagtt acttgcggcg gttgtcagat caatgagcag tggaagcaag 3960
tacttttcta gtggtgcggc aatcatcgat tttgttatat cccagggaga ttttatatat 4020
aaccaactcg ctggtttgga tgagacagcc aagaaacatg aatcaagcta tgttgagatt 4080
cctgttcttg tagctctcag agagaagagt agtaagattg acaagcctct gcagagggaa 4140
agaaacccat ctaatggtgt gaggattaaa gaagtttctc aagttgcgga gagcgaggcc 4200
ttgacatctg atcaactggt tgatggtact gatgatgaca gaagatatgc tatactctta 4260
caagacgaag agaataggaa atctatgcaa cagcccagaa aaaacagcag ctcaggttct 4320
gcttcaaata tgttctacat taagataaat gaagatgaga ttgccaatga ttatcctctc 4380
ccatcgtact ataagacctc cgaagaagaa acagatgaac ttatacttta tgatgcttcc 4440
tatgaggttc aatctgaaca cctgcctcac aggatgcttc acaactgggc tctttataac 4500
tctgatttac gattcatatc actggaactt ctaccgatga aacaatgtga tgatattgat 4560
gtcaacattt ttgggtcagg tgtggtgact gatgataatg gaagttggat ttctttaaac 4620
gatcctgaca gcggttctca gtcacacgat cctgatggga tgtgcatatt cctcagtcaa 4680
attaaagaat ggatgattga gtttgggagc gatgatatta tctccatttc tatacgaaca 4740
gatgtggcct ggtaccgtct tgggaaacca tcaaaacttt atgccccttg gtggaaacct 4800
gttctgaaaa cagcaagggt tgggataagc attcttactt ttcttagggt ggaaagtagg 4860
gttgctaggc tttcatttgc agatgtcaca aaaagactgt ctgggttaca ggcgaatgat 4920
aaagcttaca tttcttctga ccccttggct gttgagagat atttggtcgt ccatgggcaa 4980
attattttac agctttttgc agtttatccg gacgacaatg tcaaaaggtg tccatttgtt 5040
gttggtcttg caagcaaatt ggaggatagg caccacacaa aatggatcat caagaagaag 5100
aaaatttcgc tgaaggaact gaatctgaat ccaagggcag gcatggcacc agtagcatcg 5160
aagaggaaag ctatgcaagc aacaacaact cgcctggtca acagaatttg gggagagttt 5220
tactccaatt actctccaga ggatccattg caggcgactg ctgcagaaaa tggggaggat 5280
gaggtggaag aggaaggcgg aaatggggag gaagaggttg aagaggaagg tgaaaatggt 5340
ctcacagagg acactgtacc agaacctgtt gaggttcaga agcctcatac tcctaagaaa 5400
atccgaggca gttctggaaa aagggaaata aaatgggatg gtgagagtct aggaaaaact 5460
tctgctggcg agcctctcta tcaacaagcc cttgttggag gggaaatggt ggctgtaggt 5520
ggcgctgtca ccttggaagt tgatgatcca gatgaaatgc cggccatcta ttttgtggag 5580
tacatgttcg aaagtacaga tcactgcaaa atgttacatg gtagattctt acaaagagga 5640
tctatgactg ttctggggaa tgctgctaac gagagggaac tattcctgac taatgaatgc 5700
atgactacac agctcaagga cattaaagga gtagccagtt ttgagattcg atcaaggcca 5760
tgggggcatc agtataggaa aaagaacatc actgcggata agcttgactg ggctagagca 5820
ttagaaagaa aagtaaaaga tttgccaaca gagtattact gcaaaagctt gtactcacct 5880
gagagagggg gattctttag tcttccacta agtgatattg gtcgcagttc tgggttctgc 5940
acttcatgta agataaggga ggatgaagag aagaggtcta caattaaact aaatgtttca 6000
aagacaggct ttttcatcaa tgggattgag tattctgttg aggattttgt ctatgtcaac 6060
cctgactcta ttggtgggtt gaaggagggt agtaaaactt cttttaagtc tgggcgaaac 6120
attgggttaa gagcgtatgt tgtttgccaa ttgctggaaa ttgttccaaa ggaatctaga 6180
aaggctgatt tgggttcctt tgatgttaaa gtgagaaggt tttataggcc tgaggatgtt 6240
tctgcagaga aggcctatgc ttcagacatc caagaattgt atttcagcca ggacacagtt 6300
gttctccctc caggtgctct agagggaaaa tgtgaagtaa gaaagaaaag tgatatgccc 6360
ttatcccgtg aatatccaat atcagaccat attttcttct gtgatctttt ctttgacacc 6420
tccaaaggtt ctctcaagca gctgcccgcc aatatgaagc caaagttctc tactattaag 6480
gacgacacac ttttaagaaa gaaaaaggga aagggagtag agagtgaaat tgagtctgag 6540
attgtcaagc ctgttgagcc acctaaagag attcgtctgg ctactctaga tatttttgct 6600
ggttgtggtg gcctgtctca tggactgaaa aaggcgggtg tatctgatgc aaagtgggcg 6660
attgagtatg aagagccagc tgggcaggct tttaaacaaa accatcctga gtcaacagtt 6720
tttgttgaca actgcaatgt gattcttagg gctataatgg agaaaggtgg agatcaagat 6780
gattgtgtct ctactacaga ggcaaatgaa ttagcagcta aactaactga ggagcagaag 6840
agtactctgc cactgcctgg tcaagtggac ttcatcaatg gtggacctcc atgtcaggga 6900
ttttctggta tgaacaggtt caaccaaagc tcttggagta aagttcagtg tgaaatgata 6960
ttagcattct tgtcctttgc tgactatttc cggccaaggt attttcttct ggagaacgtg 7020
aggacctttg tgtcattcaa taaagggcag acatttcagc ttactttggc ttcccttctc 7080
gaaatgggtt accaggtgag atttggaatc ctggaggccg gtgcatatgg agtatcccaa 7140
tctcgtaaac gagctttcat ttgggctgct gcaccagaag aagttctccc tgaatggcct 7200
gagccgatgc atgtctttgg tgttccaaag ttgaaaatct cactatctca aggtttacat 7260
tatgctgctg ttcgtagtac tgcacttggt gcccctttcc gtccaatcac cgtgagagac 7320
acaattggtg atcttccatc agtagaaaac ggagactcta ggacaaacaa agagtataaa 7380
gaggttgcag tctcgtggtt ccaaaaggag ataagaggaa acacgattgc tctcactgat 7440
catatctgca aggctatgaa tgagcttaac ctcattcgat gcaaattaat cccaactagg 7500
cctggggctg attggcatga cttgccaaag agaaaggtta cgttatctga tgggcgcgta 7560
gaagaaatga ttcctttttg tctcccaaac acagctgagc gccacaacgg ttggaaggga 7620
ctatatggga gattagattg gcaaggaaac tttccgactt ccgtcacgga tcctcagccc 7680
atgggtaagg ttggaatgtg ctttcatcct gaacagcaca gaatccttac agtccgtgaa 7740
tgcgcccgat ctcaggggtt tccggatagc tacgagtttg cagggaacat aaatcacaag 7800
cacaggcaga ttgggaatgc agtccctcca ccattggcat ttgctctagg tcgtaagctc 7860
aaagaagccc tacatctcaa gaagtctcct caacaccaac cctag 7905
<210> 24
<211> 3339
<212> DNA
<213> RB-vTALE-AtSRDX核苷酸序列
<400> 24
atggacccca ttcgttcgcg cacgccaagt cctgcccgcg agcttctgcc cggaccccaa 60
ccggataggg ttcagccgac tgcagatcgg gggggggctc cgcctgctgg cggccccctg 120
gatggcttgc ccgctcggcg gacgatgtcc cggacccggc tgccatctcc ccctgcgccc 180
tcgcctgcgt tctcggcggg cagcttcaac gatctgctcc gtcagttcga tccgtcgctt 240
cttgatacat cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc 300
ccagcagagt gggatgaggt gcaatcgggt ctgcgtgcag ccgatgaccc gccacccacc 360
gtgcgtgtcg ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt 420
gcggcgcaac cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac 480
agtcagcagc agcaagagaa gatcaaatcg aaggtgcgtt cgacagtggc gcagcaccac 540
gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg 600
gcagcgttag ggaccgtcgc tgtcaagtat cagcacataa tcacggcgtt gccagaggcg 660
acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc 720
ttgctcacga aggcggggga gttgagaggt ccgccgttac agttggacac aggccaactt 780
ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc atcgcgcaat 840
gcactgacgg gtgcccccct gaaccttact ccagatcaag ttgtggcaat cgcttcaaat 900
atcggaggga aacaggcctt ggaaaccgtg caacgtcttc ttcctgtgtt gtgtcaagct 960
catggactca ctccagatca agttgtggct atcgctagta acggtggtgg aaaacaggcg 1020
ttagaaactg tccaaaggct ccttcccgtg ttgtgtcaag ctcatggcct cactccagat 1080
caagttgtgg caatcgcttc aaatatcgga gggaaacagg ccttggaaac cgtgcaacgt 1140
cttcttcctg tgttgtgtca agctcatgga ttaactccag atcaagttgt ggctatcgct 1200
agtaacggtg gtggaaaaca ggcgttagaa actgtccaaa ggctccttcc cgtgttgtgt 1260
caagctcatg gcttaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 1320
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggactcact 1380
ccagatcaag ttgtggctat cgctagtaac ggtggtggaa aacaggcgtt agaaactgtc 1440
caaaggctcc ttcccgtgtt gtgtcaagct catggcctca ctccagatca agttgtggct 1500
atcgctagta acggtggtgg aaaacaggcg ttagaaactg tccaaaggct ccttcccgtg 1560
ttgtgtcaag ctcatggact cactccagat caagttgtgg caatagcctc taacaatgga 1620
ggtaaacaag cacttgagac tgttcagaga ttgttaccgg tgttgtgtca agctcatggc 1680
ctcactccag atcaagttgt ggcaatagcc tctaacaatg gaggtaaaca agcacttgag 1740
actgttcaga gattgttacc ggtgttgtgt caagctcatg gattaactcc agatcaagtt 1800
gtggcaatcg cttcaaatat cggagggaaa caggccttgg aaaccgtgca acgtcttctt 1860
cctgtgttgt gtcaagctca tggtctcact ccagatcaag ttgtggccat tgcatctcat 1920
gatggtggta agcaagctct cgaaacagtc caaagacttc tcccagtgtt gtgtcaagct 1980
catggattaa ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 2040
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggtct cactccagat 2100
caagttgtgg ccattgcatc tcatgatggt ggtaagcaag ctctcgaaac agtccaaaga 2160
cttctcccag tgttgtgtca agctcatgga ctcactccag atcaagttgt ggccattgca 2220
tctcatgatg gtggtaagca agctctcgaa acagtccaaa gacttctccc agtgttgtgt 2280
caagctcatg gattaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 2340
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggtttaact 2400
ccagatcaag ttgtggcaat cgcttcaaat atcggaggga aacaggcctt ggaaaccgtg 2460
caacgtcttc ttcctgtgtt gtgtcaagct catggtctca ctccagatca agttgtggca 2520
atagcctcta acaatggagg taaacaagca cttgagagca ttgttgccca gttatctcgc 2580
cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc 2640
ggacgtcctg ccctggatgc agtgaaaaag ggattgccgc acgcgccgga attgatcaga 2700
agaatcaatc gccgtattcc cgaacgcacg tcccatcgcg ttgccgacct cgcgcacgtg 2760
gtgcgcgtgc ttggtttttt ccagagccac tcccacccag cgcaagcatt cgatgacgcc 2820
atgacgcagt tcgggatgag caggcacggg ttggtacagc tctttcgcag agtgggcgtc 2880
accgaattcg aagcccgctg cggaacgctc cccccagcct cgcagcgttg ggaccgtatc 2940
ctccaggcat cagggatgaa aagggccaaa ccgtccccta cttcagctca aacgccggat 3000
caggcgtctt tgcatgcatt cgccgattcg ctggagcgtg accttgatgc gcccagccca 3060
atgcacgagg gagatcagac gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct 3120
gtcaccggcc cctccacaca gcaatctttc gaggtgcgcg ttcccgaaca gcgcgatgcg 3180
ctgcatttgc ccctcagctg gagggtaaaa cgcccgcgta ccaggatcgg gggcggcctc 3240
ccggatcctg gtacgcccat cgctgccgac ctggcagcgt ccagcaccgt gatgtggtct 3300
agtcttgatc tggatctgga actccgtttg ggtttcgct 3339
<210> 25
<211> 5256
<212> DNA
<213> RB-vTALE-AtSUVH2核苷酸序列
<400> 25
atggacccca ttcgttcgcg cacgccaagt cctgcccgcg agcttctgcc cggaccccaa 60
ccggataggg ttcagccgac tgcagatcgg gggggggctc cgcctgctgg cggccccctg 120
gatggcttgc ccgctcggcg gacgatgtcc cggacccggc tgccatctcc ccctgcgccc 180
tcgcctgcgt tctcggcggg cagcttcaac gatctgctcc gtcagttcga tccgtcgctt 240
cttgatacat cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc 300
ccagcagagt gggatgaggt gcaatcgggt ctgcgtgcag ccgatgaccc gccacccacc 360
gtgcgtgtcg ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt 420
gcggcgcaac cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac 480
agtcagcagc agcaagagaa gatcaaatcg aaggtgcgtt cgacagtggc gcagcaccac 540
gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg 600
gcagcgttag ggaccgtcgc tgtcaagtat cagcacataa tcacggcgtt gccagaggcg 660
acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc 720
ttgctcacga aggcggggga gttgagaggt ccgccgttac agttggacac aggccaactt 780
ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc atcgcgcaat 840
gcactgacgg gtgcccccct gaaccttact ccagatcaag ttgtggcaat cgcttcaaat 900
atcggaggga aacaggcctt ggaaaccgtg caacgtcttc ttcctgtgtt gtgtcaagct 960
catggactca ctccagatca agttgtggct atcgctagta acggtggtgg aaaacaggcg 1020
ttagaaactg tccaaaggct ccttcccgtg ttgtgtcaag ctcatggcct cactccagat 1080
caagttgtgg caatcgcttc aaatatcgga gggaaacagg ccttggaaac cgtgcaacgt 1140
cttcttcctg tgttgtgtca agctcatgga ttaactccag atcaagttgt ggctatcgct 1200
agtaacggtg gtggaaaaca ggcgttagaa actgtccaaa ggctccttcc cgtgttgtgt 1260
caagctcatg gcttaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 1320
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggactcact 1380
ccagatcaag ttgtggctat cgctagtaac ggtggtggaa aacaggcgtt agaaactgtc 1440
caaaggctcc ttcccgtgtt gtgtcaagct catggcctca ctccagatca agttgtggct 1500
atcgctagta acggtggtgg aaaacaggcg ttagaaactg tccaaaggct ccttcccgtg 1560
ttgtgtcaag ctcatggact cactccagat caagttgtgg caatagcctc taacaatgga 1620
ggtaaacaag cacttgagac tgttcagaga ttgttaccgg tgttgtgtca agctcatggc 1680
ctcactccag atcaagttgt ggcaatagcc tctaacaatg gaggtaaaca agcacttgag 1740
actgttcaga gattgttacc ggtgttgtgt caagctcatg gattaactcc agatcaagtt 1800
gtggcaatcg cttcaaatat cggagggaaa caggccttgg aaaccgtgca acgtcttctt 1860
cctgtgttgt gtcaagctca tggtctcact ccagatcaag ttgtggccat tgcatctcat 1920
gatggtggta agcaagctct cgaaacagtc caaagacttc tcccagtgtt gtgtcaagct 1980
catggattaa ctccagatca agttgtggca atcgcttcaa atatcggagg gaaacaggcc 2040
ttggaaaccg tgcaacgtct tcttcctgtg ttgtgtcaag ctcatggtct cactccagat 2100
caagttgtgg ccattgcatc tcatgatggt ggtaagcaag ctctcgaaac agtccaaaga 2160
cttctcccag tgttgtgtca agctcatgga ctcactccag atcaagttgt ggccattgca 2220
tctcatgatg gtggtaagca agctctcgaa acagtccaaa gacttctccc agtgttgtgt 2280
caagctcatg gattaactcc agatcaagtt gtggcaatcg cttcaaatat cggagggaaa 2340
caggccttgg aaaccgtgca acgtcttctt cctgtgttgt gtcaagctca tggtttaact 2400
ccagatcaag ttgtggcaat cgcttcaaat atcggaggga aacaggcctt ggaaaccgtg 2460
caacgtcttc ttcctgtgtt gtgtcaagct catggtctca ctccagatca agttgtggca 2520
atagcctcta acaatggagg taaacaagca cttgagagca ttgttgccca gttatctcgc 2580
cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc 2640
ggacgtcctg ccctggatgc agtgaaaaag ggattgccgc acgcgccgga attgatcaga 2700
agaatcaatc gccgtattcc cgaacgcacg tcccatcgcg ttgccgacct cgcgcacgtg 2760
gtgcgcgtgc ttggtttttt ccagagccac tcccacccag cgcaagcatt cgatgacgcc 2820
atgacgcagt tcgggatgag caggcacggg ttggtacagc tctttcgcag agtgggcgtc 2880
accgaattcg aagcccgctg cggaacgctc cccccagcct cgcagcgttg ggaccgtatc 2940
ctccaggcat cagggatgaa aagggccaaa ccgtccccta cttcagctca aacgccggat 3000
caggcgtctt tgcatgcatt cgccgattcg ctggagcgtg accttgatgc gcccagccca 3060
atgcacgagg gagatcagac gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct 3120
gtcaccggcc cctccacaca gcaatctttc gaggtgcgcg ttcccgaaca gcgcgatgcg 3180
ctgcatttgc ccctcagctg gagggtaaaa cgcccgcgta ccaggatcgg gggcggcctc 3240
ccggatcctg gtacgcccat cgctgccgac ctggcagcgt ccagcaccgt gatgtggtct 3300
agtagtacat tgttaccatt tcctgacctc aacctcatgc cggattctca atcctccacc 3360
gccggaacca cagccggcga cactgtagtc accggaaagt tagaagtgaa atcggaacca 3420
attgaggaat ggcaaactcc accatcgtct acctccgatc aatcagccaa taccgatctc 3480
atcgccgagt ttattcgtat ctcagagctc ttccgctcgg cattcaagcc actgcaggtc 3540
aaaggattag acggagtctc cgtatacgga ttggattctg gtgccatcgt tgctgtaccg 3600
gagaaagaaa accgggaatt gattgaaccg cctccaggat ttaaggataa tcgagtctcc 3660
accgtcgttg tctcgccgaa attcgagaga ccgagagagc tagcgagaat tgcaattcta 3720
ggtcatgaac aacggaagga actccgacaa gtcatgaaac gaactaggat gacttatgag 3780
tctcttcgga ttcatctaat ggcggagagt atgaagaatc atgtacttgg tcaagggcgt 3840
agacggagga gcgatatggc ggctgcgtat ataatgaggg atcggggact gtggttgaac 3900
tatgataagc acatagttgg tccagtcaca ggggttgaag taggggatat attcttctac 3960
cgtatggaat tgtgtgtgct gggcttacac ggccagacac aagccgggat tgattgttta 4020
acggctgaac ggagcgccac tggagagcct atagctacca gcattgttgt ctcaggtggt 4080
tatgaggatg atgaagatac aggagatgtt ttggtttata ctggtcacgg tggtcaggat 4140
catcaacaca agcagtgtga taaccagagg ctagtaggtg ggaatctggg aatggagaga 4200
agtatgcact acggtatcga ggtacgtgtg attagaggta ttaagtatga gaacagtata 4260
agctcgaagg tgtatgtata tgatggtttg tataagatag tagattggtg gtttgctgtg 4320
gggaagtctg gttttggggt ttttaaattt aggttagtaa gaattgaagg gcaaccgatg 4380
atgggcagtg cggtaatgag gtttgcacaa actcttagaa ataagccatc gatggttagg 4440
cctactggtt atgttagctt tgacctttct aacaagaagg agaatgtacc cgtgtttcta 4500
tataatgatg tagatggtga tcaagaaccg agacattatg agtacattgc aaaagctgtc 4560
tttcctcctg gaatttttgg tcaggggggg atcagcagga ctggctgcga gtgtaagctc 4620
tcttgtactg atgattgtct ctgtgcaagg aagaacggtg gtgagtttgc atatgatgat 4680
aacgggcatc tattgaaagg aaagcatgtg gtatttgaat gtggggaatt ctgcacttgt 4740
ggtccgagct gtaagagccg tgtgacacag aagggattga ggaacaggct agaggttttc 4800
agatcgaagg aaaccggttg gggtgtcagg acacttgatc taattgaagc tggtgctttc 4860
atatgcgaat atgcaggtgt agttgtcacg agacttcaag ccgagattct gtcaatgaat 4920
ggggatgtta tggtctatcc tggtcggttc acagaccaat ggcgtaactg gggtgattta 4980
tctcaagtat acccagattt tgttaggccg aattatccat ctctccctcc actggatttc 5040
tcaatggatg tgtcaaggat gaggaacgtg gcttgctaca ttagtcacag caaagaacca 5100
aatgtgatgg tgcaatttgt tttgcatgac cacaatcacc tgatgttccc ccgtgtgatg 5160
ctttttgcgc tggagaatat ctcaccgttg gccgagctaa gcctggatta cggcttggca 5220
gatgaagtga acggcaagct cgccatctgc aactag 5256
<210> 26
<211> 1724
<212> PRT
<213> RB-vTALE-AtKYP氨基酸序列
<400> 26
Met Asp Pro Ile Arg Ser Arg Thr Pro Ser Pro Ala Arg Glu Leu Leu
1 5 10 15
Pro Gly Pro Gln Pro Asp Arg Val Gln Pro Thr Ala Asp Arg Gly Gly
20 25 30
Ala Pro Pro Ala Gly Gly Pro Leu Asp Gly Leu Pro Ala Arg Arg Thr
35 40 45
Met Ser Arg Thr Arg Leu Pro Ser Pro Pro Ala Pro Ser Pro Ala Phe
50 55 60
Ser Ala Gly Ser Phe Asn Asp Leu Leu Arg Gln Phe Asp Pro Ser Leu
65 70 75 80
Leu Asp Thr Ser Leu Leu Asp Ser Met Pro Ala Val Gly Thr Pro His
85 90 95
Thr Ala Ala Ala Pro Ala Glu Trp Asp Glu Val Gln Ser Gly Leu Arg
100 105 110
Ala Ala Asp Asp Pro Pro Pro Thr Val Arg Val Ala Val Thr Ala Ala
115 120 125
Arg Pro Pro Arg Ala Lys Pro Ala Pro Arg Arg Arg Ala Ala Gln Pro
130 135 140
Ser Asp Ala Ser Pro Ala Ala Gln Val Asp Leu Arg Thr Leu Gly Tyr
145 150 155 160
Ser Gln Gln Gln Gln Glu Lys Ile Lys Ser Lys Val Arg Ser Thr Val
165 170 175
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
180 185 190
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
195 200 205
Lys Tyr Gln His Ile Ile Thr Ala Leu Pro Glu Ala Thr His Glu Asp
210 215 220
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
225 230 235 240
Leu Leu Thr Lys Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
245 250 255
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
260 265 270
Glu Ala Val His Ala Ser Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
275 280 285
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys
290 295 300
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
305 310 315 320
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly
325 330 335
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
340 345 350
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn
355 360 365
Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
370 375 380
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
385 390 395 400
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
405 410 415
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
420 425 430
Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
435 440 445
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
450 455 460
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
465 470 475 480
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
485 490 495
Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu
500 505 510
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
515 520 525
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
530 535 540
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
545 550 555 560
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
565 570 575
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
580 585 590
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly
595 600 605
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
610 615 620
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His
625 630 635 640
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
645 650 655
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
660 665 670
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
675 680 685
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
690 695 700
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
705 710 715 720
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
725 730 735
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
740 745 750
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
755 760 765
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
770 775 780
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
785 790 795 800
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
805 810 815
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
820 825 830
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
835 840 845
Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala
850 855 860
Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly
865 870 875 880
Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Pro His Ala Pro
885 890 895
Glu Leu Ile Arg Arg Ile Asn Arg Arg Ile Pro Glu Arg Thr Ser His
900 905 910
Arg Val Ala Asp Leu Ala His Val Val Arg Val Leu Gly Phe Phe Gln
915 920 925
Ser His Ser His Pro Ala Gln Ala Phe Asp Asp Ala Met Thr Gln Phe
930 935 940
Gly Met Ser Arg His Gly Leu Val Gln Leu Phe Arg Arg Val Gly Val
945 950 955 960
Thr Glu Phe Glu Ala Arg Cys Gly Thr Leu Pro Pro Ala Ser Gln Arg
965 970 975
Trp Asp Arg Ile Leu Gln Ala Ser Gly Met Lys Arg Ala Lys Pro Ser
980 985 990
Pro Thr Ser Ala Gln Thr Pro Asp Gln Ala Ser Leu His Ala Phe Ala
995 1000 1005
Asp Ser Leu Glu Arg Asp Leu Asp Ala Pro Ser Pro Met His Glu
1010 1015 1020
Gly Asp Gln Thr Arg Ala Ser Ser Arg Lys Arg Ser Arg Ser Asp
1025 1030 1035
Arg Ala Val Thr Gly Pro Ser Thr Gln Gln Ser Phe Glu Val Arg
1040 1045 1050
Val Pro Glu Gln Arg Asp Ala Leu His Leu Pro Leu Ser Trp Arg
1055 1060 1065
Val Lys Arg Pro Arg Thr Arg Ile Gly Gly Gly Leu Pro Asp Pro
1070 1075 1080
Gly Thr Pro Ile Ala Ala Asp Leu Ala Ala Ser Ser Thr Val Met
1085 1090 1095
Trp Ser Ser Ala Gly Lys Arg Lys Arg Ala Asn Ala Pro Asp Gln
1100 1105 1110
Thr Glu Arg Arg Ser Ser Val Arg Val Gln Lys Val Arg Gln Lys
1115 1120 1125
Ala Leu Asp Glu Lys Ala Arg Leu Val Gln Glu Arg Val Lys Leu
1130 1135 1140
Leu Ser Asp Arg Lys Ser Glu Ile Cys Val Asp Asp Thr Glu Leu
1145 1150 1155
His Glu Lys Glu Glu Glu Asn Val Asp Gly Ser Pro Lys Arg Arg
1160 1165 1170
Ser Pro Pro Lys Leu Thr Ala Met Gln Lys Gly Lys Gln Lys Leu
1175 1180 1185
Ser Val Ser Leu Asn Gly Lys Asp Val Asn Leu Glu Pro His Leu
1190 1195 1200
Lys Val Thr Lys Cys Leu Arg Leu Phe Asn Lys Gln Tyr Leu Leu
1205 1210 1215
Cys Val Gln Ala Lys Leu Ser Arg Pro Asp Leu Lys Gly Val Thr
1220 1225 1230
Glu Met Ile Lys Ala Lys Ala Ile Leu Tyr Pro Arg Lys Ile Ile
1235 1240 1245
Gly Asp Leu Pro Gly Ile Asp Val Gly His Arg Phe Phe Ser Arg
1250 1255 1260
Ala Glu Met Cys Ala Val Gly Phe His Asn His Trp Leu Asn Gly
1265 1270 1275
Ile Asp Tyr Met Ser Met Glu Tyr Glu Lys Glu Tyr Ser Asn Tyr
1280 1285 1290
Lys Leu Pro Leu Ala Val Ser Ile Val Met Ser Gly Gln Tyr Glu
1295 1300 1305
Asp Asp Leu Asp Asn Ala Asp Thr Val Thr Tyr Thr Gly Gln Gly
1310 1315 1320
Gly His Asn Leu Thr Gly Asn Lys Arg Gln Ile Lys Asp Gln Leu
1325 1330 1335
Leu Glu Arg Gly Asn Leu Ala Leu Lys His Cys Cys Glu Tyr Asn
1340 1345 1350
Val Pro Val Arg Val Thr Arg Gly His Asn Cys Lys Ser Ser Tyr
1355 1360 1365
Thr Lys Arg Val Tyr Thr Tyr Asp Gly Leu Tyr Lys Val Glu Lys
1370 1375 1380
Phe Trp Ala Gln Lys Gly Val Ser Gly Phe Thr Val Tyr Lys Tyr
1385 1390 1395
Arg Leu Lys Arg Leu Glu Gly Gln Pro Glu Leu Thr Thr Asp Gln
1400 1405 1410
Val Asn Phe Val Ala Gly Arg Ile Pro Thr Ser Thr Ser Glu Ile
1415 1420 1425
Glu Gly Leu Val Cys Glu Asp Ile Ser Gly Gly Leu Glu Phe Lys
1430 1435 1440
Gly Ile Pro Ala Thr Asn Arg Val Asp Asp Ser Pro Val Ser Pro
1445 1450 1455
Thr Ser Gly Phe Thr Tyr Ile Lys Ser Leu Ile Ile Glu Pro Asn
1460 1465 1470
Val Ile Ile Pro Lys Ser Ser Thr Gly Cys Asn Cys Arg Gly Ser
1475 1480 1485
Cys Thr Asp Ser Lys Lys Cys Ala Cys Ala Lys Leu Asn Gly Gly
1490 1495 1500
Asn Phe Pro Tyr Val Asp Leu Asn Asp Gly Arg Leu Ile Glu Ser
1505 1510 1515
Arg Asp Val Val Phe Glu Cys Gly Pro His Cys Gly Cys Gly Pro
1520 1525 1530
Lys Cys Val Asn Arg Thr Ser Gln Lys Arg Leu Arg Phe Asn Leu
1535 1540 1545
Glu Val Phe Arg Ser Ala Lys Lys Gly Trp Ala Val Arg Ser Trp
1550 1555 1560
Glu Tyr Ile Pro Ala Gly Ser Pro Val Cys Glu Tyr Ile Gly Val
1565 1570 1575
Val Arg Arg Thr Ala Asp Val Asp Thr Ile Ser Asp Asn Glu Tyr
1580 1585 1590
Ile Phe Glu Ile Asp Cys Gln Gln Thr Met Gln Gly Leu Gly Gly
1595 1600 1605
Arg Gln Arg Arg Leu Arg Asp Val Ala Val Pro Met Asn Asn Gly
1610 1615 1620
Val Ser Gln Ser Ser Glu Asp Glu Asn Ala Pro Glu Phe Cys Ile
1625 1630 1635
Asp Ala Gly Ser Thr Gly Asn Phe Ala Arg Phe Ile Asn His Ser
1640 1645 1650
Cys Glu Pro Asn Leu Phe Val Gln Cys Val Leu Ser Ser His Gln
1655 1660 1665
Asp Ile Arg Leu Ala Arg Val Val Leu Phe Ala Ala Asp Asn Ile
1670 1675 1680
Ser Pro Met Gln Glu Leu Thr Tyr Asp Tyr Gly Tyr Ala Leu Asp
1685 1690 1695
Ser Val His Gly Pro Asp Gly Lys Val Lys Gln Leu Ala Cys Tyr
1700 1705 1710
Cys Gly Ala Leu Asn Cys Arg Lys Arg Leu Tyr
1715 1720
<210> 27
<211> 1569
<212> PRT
<213> RB-vTALE-AtHDAC19氨基酸序列
<400> 27
Met Asp Pro Ile Arg Ser Arg Thr Pro Ser Pro Ala Arg Glu Leu Leu
1 5 10 15
Pro Gly Pro Gln Pro Asp Arg Val Gln Pro Thr Ala Asp Arg Gly Gly
20 25 30
Ala Pro Pro Ala Gly Gly Pro Leu Asp Gly Leu Pro Ala Arg Arg Thr
35 40 45
Met Ser Arg Thr Arg Leu Pro Ser Pro Pro Ala Pro Ser Pro Ala Phe
50 55 60
Ser Ala Gly Ser Phe Asn Asp Leu Leu Arg Gln Phe Asp Pro Ser Leu
65 70 75 80
Leu Asp Thr Ser Leu Leu Asp Ser Met Pro Ala Val Gly Thr Pro His
85 90 95
Thr Ala Ala Ala Pro Ala Glu Trp Asp Glu Val Gln Ser Gly Leu Arg
100 105 110
Ala Ala Asp Asp Pro Pro Pro Thr Val Arg Val Ala Val Thr Ala Ala
115 120 125
Arg Pro Pro Arg Ala Lys Pro Ala Pro Arg Arg Arg Ala Ala Gln Pro
130 135 140
Ser Asp Ala Ser Pro Ala Ala Gln Val Asp Leu Arg Thr Leu Gly Tyr
145 150 155 160
Ser Gln Gln Gln Gln Glu Lys Ile Lys Ser Lys Val Arg Ser Thr Val
165 170 175
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
180 185 190
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
195 200 205
Lys Tyr Gln His Ile Ile Thr Ala Leu Pro Glu Ala Thr His Glu Asp
210 215 220
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
225 230 235 240
Leu Leu Thr Lys Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
245 250 255
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
260 265 270
Glu Ala Val His Ala Ser Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
275 280 285
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys
290 295 300
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
305 310 315 320
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly
325 330 335
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
340 345 350
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn
355 360 365
Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
370 375 380
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
385 390 395 400
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
405 410 415
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
420 425 430
Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
435 440 445
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
450 455 460
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
465 470 475 480
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
485 490 495
Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu
500 505 510
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
515 520 525
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
530 535 540
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
545 550 555 560
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
565 570 575
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
580 585 590
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly
595 600 605
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
610 615 620
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His
625 630 635 640
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
645 650 655
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
660 665 670
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
675 680 685
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
690 695 700
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
705 710 715 720
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
725 730 735
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
740 745 750
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
755 760 765
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
770 775 780
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
785 790 795 800
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
805 810 815
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
820 825 830
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
835 840 845
Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala
850 855 860
Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly
865 870 875 880
Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Pro His Ala Pro
885 890 895
Glu Leu Ile Arg Arg Ile Asn Arg Arg Ile Pro Glu Arg Thr Ser His
900 905 910
Arg Val Ala Asp Leu Ala His Val Val Arg Val Leu Gly Phe Phe Gln
915 920 925
Ser His Ser His Pro Ala Gln Ala Phe Asp Asp Ala Met Thr Gln Phe
930 935 940
Gly Met Ser Arg His Gly Leu Val Gln Leu Phe Arg Arg Val Gly Val
945 950 955 960
Thr Glu Phe Glu Ala Arg Cys Gly Thr Leu Pro Pro Ala Ser Gln Arg
965 970 975
Trp Asp Arg Ile Leu Gln Ala Ser Gly Met Lys Arg Ala Lys Pro Ser
980 985 990
Pro Thr Ser Ala Gln Thr Pro Asp Gln Ala Ser Leu His Ala Phe Ala
995 1000 1005
Asp Ser Leu Glu Arg Asp Leu Asp Ala Pro Ser Pro Met His Glu
1010 1015 1020
Gly Asp Gln Thr Arg Ala Ser Ser Arg Lys Arg Ser Arg Ser Asp
1025 1030 1035
Arg Ala Val Thr Gly Pro Ser Thr Gln Gln Ser Phe Glu Val Arg
1040 1045 1050
Val Pro Glu Gln Arg Asp Ala Leu His Leu Pro Leu Ser Trp Arg
1055 1060 1065
Val Lys Arg Pro Arg Thr Arg Ile Gly Gly Gly Leu Pro Asp Pro
1070 1075 1080
Gly Thr Pro Ile Ala Ala Asp Leu Ala Ala Ser Ser Thr Val Met
1085 1090 1095
Trp Ser Ser Asp Thr Gly Gly Asn Ser Leu Ala Ser Gly Pro Asp
1100 1105 1110
Gly Val Lys Arg Lys Val Cys Tyr Phe Tyr Asp Pro Glu Val Gly
1115 1120 1125
Asn Tyr Tyr Tyr Gly Gln Gly His Pro Met Lys Pro His Arg Ile
1130 1135 1140
Arg Met Thr His Ala Leu Leu Ala His Tyr Gly Leu Leu Gln His
1145 1150 1155
Met Gln Val Leu Lys Pro Phe Pro Ala Arg Asp Arg Asp Leu Cys
1160 1165 1170
Arg Phe His Ala Asp Asp Tyr Val Ser Phe Leu Arg Ser Ile Thr
1175 1180 1185
Pro Glu Thr Gln Gln Asp Gln Ile Arg Gln Leu Lys Arg Phe Asn
1190 1195 1200
Val Gly Glu Asp Cys Pro Val Phe Asp Gly Leu Tyr Ser Phe Cys
1205 1210 1215
Gln Thr Tyr Ala Gly Gly Ser Val Gly Gly Ser Val Lys Leu Asn
1220 1225 1230
His Gly Leu Cys Asp Ile Ala Ile Asn Trp Ala Gly Gly Leu His
1235 1240 1245
His Ala Lys Lys Cys Glu Ala Ser Gly Phe Cys Tyr Val Asn Asp
1250 1255 1260
Ile Val Leu Ala Ile Leu Glu Leu Leu Lys Gln His Glu Arg Val
1265 1270 1275
Leu Tyr Val Asp Ile Asp Ile His His Gly Asp Gly Val Glu Glu
1280 1285 1290
Ala Phe Tyr Ala Thr Asp Arg Val Met Thr Val Ser Phe His Lys
1295 1300 1305
Phe Gly Asp Tyr Phe Pro Gly Thr Gly His Ile Gln Asp Ile Gly
1310 1315 1320
Tyr Gly Ser Gly Lys Tyr Tyr Ser Leu Asn Val Pro Leu Asp Asp
1325 1330 1335
Gly Ile Asp Asp Glu Ser Tyr His Leu Leu Phe Lys Pro Ile Met
1340 1345 1350
Gly Lys Val Met Glu Ile Phe Arg Pro Gly Ala Val Val Leu Gln
1355 1360 1365
Cys Gly Ala Asp Ser Leu Ser Gly Asp Arg Leu Gly Cys Phe Asn
1370 1375 1380
Leu Ser Ile Lys Gly His Ala Glu Cys Val Lys Phe Met Arg Ser
1385 1390 1395
Phe Asn Val Pro Leu Leu Leu Leu Gly Gly Gly Gly Tyr Thr Ile
1400 1405 1410
Arg Asn Val Ala Arg Cys Trp Cys Tyr Glu Thr Gly Val Ala Leu
1415 1420 1425
Gly Val Glu Val Glu Asp Lys Met Pro Glu His Glu Tyr Tyr Glu
1430 1435 1440
Tyr Phe Gly Pro Asp Tyr Thr Leu His Val Ala Pro Ser Asn Met
1445 1450 1455
Glu Asn Lys Asn Ser Arg Gln Met Leu Glu Glu Ile Arg Asn Asp
1460 1465 1470
Leu Leu His Asn Leu Ser Lys Leu Gln His Ala Pro Ser Val Pro
1475 1480 1485
Phe Gln Glu Arg Pro Pro Asp Thr Glu Thr Pro Glu Val Asp Glu
1490 1495 1500
Asp Gln Glu Asp Gly Asp Lys Arg Trp Asp Pro Asp Ser Asp Met
1505 1510 1515
Asp Val Asp Asp Asp Arg Lys Pro Ile Pro Ser Arg Val Lys Arg
1520 1525 1530
Glu Ala Val Glu Pro Asp Thr Lys Asp Lys Asp Gly Leu Lys Gly
1535 1540 1545
Ile Met Glu Arg Gly Lys Gly Cys Glu Val Glu Val Asp Glu Ser
1550 1555 1560
Gly Ser Thr Lys Val Lys
1565
<210> 28
<211> 2634
<212> PRT
<213> RB-vTALE-AtMET1氨基酸序列
<400> 28
Met Asp Pro Ile Arg Ser Arg Thr Pro Ser Pro Ala Arg Glu Leu Leu
1 5 10 15
Pro Gly Pro Gln Pro Asp Arg Val Gln Pro Thr Ala Asp Arg Gly Gly
20 25 30
Ala Pro Pro Ala Gly Gly Pro Leu Asp Gly Leu Pro Ala Arg Arg Thr
35 40 45
Met Ser Arg Thr Arg Leu Pro Ser Pro Pro Ala Pro Ser Pro Ala Phe
50 55 60
Ser Ala Gly Ser Phe Asn Asp Leu Leu Arg Gln Phe Asp Pro Ser Leu
65 70 75 80
Leu Asp Thr Ser Leu Leu Asp Ser Met Pro Ala Val Gly Thr Pro His
85 90 95
Thr Ala Ala Ala Pro Ala Glu Trp Asp Glu Val Gln Ser Gly Leu Arg
100 105 110
Ala Ala Asp Asp Pro Pro Pro Thr Val Arg Val Ala Val Thr Ala Ala
115 120 125
Arg Pro Pro Arg Ala Lys Pro Ala Pro Arg Arg Arg Ala Ala Gln Pro
130 135 140
Ser Asp Ala Ser Pro Ala Ala Gln Val Asp Leu Arg Thr Leu Gly Tyr
145 150 155 160
Ser Gln Gln Gln Gln Glu Lys Ile Lys Ser Lys Val Arg Ser Thr Val
165 170 175
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
180 185 190
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
195 200 205
Lys Tyr Gln His Ile Ile Thr Ala Leu Pro Glu Ala Thr His Glu Asp
210 215 220
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
225 230 235 240
Leu Leu Thr Lys Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
245 250 255
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
260 265 270
Glu Ala Val His Ala Ser Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
275 280 285
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys
290 295 300
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
305 310 315 320
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly
325 330 335
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
340 345 350
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn
355 360 365
Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
370 375 380
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
385 390 395 400
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
405 410 415
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
420 425 430
Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
435 440 445
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
450 455 460
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
465 470 475 480
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
485 490 495
Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu
500 505 510
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
515 520 525
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
530 535 540
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
545 550 555 560
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
565 570 575
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
580 585 590
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly
595 600 605
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
610 615 620
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His
625 630 635 640
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
645 650 655
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
660 665 670
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
675 680 685
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
690 695 700
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
705 710 715 720
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
725 730 735
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
740 745 750
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
755 760 765
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
770 775 780
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
785 790 795 800
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
805 810 815
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
820 825 830
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
835 840 845
Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala
850 855 860
Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly
865 870 875 880
Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Pro His Ala Pro
885 890 895
Glu Leu Ile Arg Arg Ile Asn Arg Arg Ile Pro Glu Arg Thr Ser His
900 905 910
Arg Val Ala Asp Leu Ala His Val Val Arg Val Leu Gly Phe Phe Gln
915 920 925
Ser His Ser His Pro Ala Gln Ala Phe Asp Asp Ala Met Thr Gln Phe
930 935 940
Gly Met Ser Arg His Gly Leu Val Gln Leu Phe Arg Arg Val Gly Val
945 950 955 960
Thr Glu Phe Glu Ala Arg Cys Gly Thr Leu Pro Pro Ala Ser Gln Arg
965 970 975
Trp Asp Arg Ile Leu Gln Ala Ser Gly Met Lys Arg Ala Lys Pro Ser
980 985 990
Pro Thr Ser Ala Gln Thr Pro Asp Gln Ala Ser Leu His Ala Phe Ala
995 1000 1005
Asp Ser Leu Glu Arg Asp Leu Asp Ala Pro Ser Pro Met His Glu
1010 1015 1020
Gly Asp Gln Thr Arg Ala Ser Ser Arg Lys Arg Ser Arg Ser Asp
1025 1030 1035
Arg Ala Val Thr Gly Pro Ser Thr Gln Gln Ser Phe Glu Val Arg
1040 1045 1050
Val Pro Glu Gln Arg Asp Ala Leu His Leu Pro Leu Ser Trp Arg
1055 1060 1065
Val Lys Arg Pro Arg Thr Arg Ile Gly Gly Gly Leu Pro Asp Pro
1070 1075 1080
Gly Thr Pro Ile Ala Ala Asp Leu Ala Ala Ser Ser Thr Val Met
1085 1090 1095
Trp Ser Ser Val Glu Asn Gly Ala Lys Ala Ala Lys Arg Lys Lys
1100 1105 1110
Arg Pro Leu Pro Glu Ile Gln Glu Val Glu Asp Val Pro Arg Thr
1115 1120 1125
Arg Arg Pro Arg Arg Ala Ala Ala Cys Thr Ser Phe Lys Glu Lys
1130 1135 1140
Ser Ile Arg Val Cys Glu Lys Ser Ala Thr Ile Glu Val Lys Lys
1145 1150 1155
Gln Gln Ile Val Glu Glu Glu Phe Leu Ala Leu Arg Leu Thr Ala
1160 1165 1170
Leu Glu Thr Asp Val Glu Asp Arg Pro Thr Arg Arg Leu Asn Asp
1175 1180 1185
Phe Val Leu Phe Asp Ser Asp Gly Val Pro Gln Pro Leu Glu Met
1190 1195 1200
Leu Glu Ile His Asp Ile Phe Val Ser Gly Ala Ile Leu Pro Ser
1205 1210 1215
Asp Val Cys Thr Asp Lys Glu Lys Glu Lys Gly Val Arg Cys Thr
1220 1225 1230
Ser Phe Gly Arg Val Glu His Trp Ser Ile Ser Gly Tyr Glu Asp
1235 1240 1245
Gly Ser Pro Val Ile Trp Ile Ser Thr Glu Leu Ala Asp Tyr Asp
1250 1255 1260
Cys Arg Lys Pro Ala Ala Ser Tyr Arg Lys Val Tyr Asp Tyr Phe
1265 1270 1275
Tyr Glu Lys Ala Arg Ala Ser Val Ala Val Tyr Lys Lys Leu Ser
1280 1285 1290
Lys Ser Ser Gly Gly Asp Pro Asp Ile Gly Leu Glu Glu Leu Leu
1295 1300 1305
Ala Ala Val Val Arg Ser Met Ser Ser Gly Ser Lys Tyr Phe Ser
1310 1315 1320
Ser Gly Ala Ala Ile Ile Asp Phe Val Ile Ser Gln Gly Asp Phe
1325 1330 1335
Ile Tyr Asn Gln Leu Ala Gly Leu Asp Glu Thr Ala Lys Lys His
1340 1345 1350
Glu Ser Ser Tyr Val Glu Ile Pro Val Leu Val Ala Leu Arg Glu
1355 1360 1365
Lys Ser Ser Lys Ile Asp Lys Pro Leu Gln Arg Glu Arg Asn Pro
1370 1375 1380
Ser Asn Gly Val Arg Ile Lys Glu Val Ser Gln Val Ala Glu Ser
1385 1390 1395
Glu Ala Leu Thr Ser Asp Gln Leu Val Asp Gly Thr Asp Asp Asp
1400 1405 1410
Arg Arg Tyr Ala Ile Leu Leu Gln Asp Glu Glu Asn Arg Lys Ser
1415 1420 1425
Met Gln Gln Pro Arg Lys Asn Ser Ser Ser Gly Ser Ala Ser Asn
1430 1435 1440
Met Phe Tyr Ile Lys Ile Asn Glu Asp Glu Ile Ala Asn Asp Tyr
1445 1450 1455
Pro Leu Pro Ser Tyr Tyr Lys Thr Ser Glu Glu Glu Thr Asp Glu
1460 1465 1470
Leu Ile Leu Tyr Asp Ala Ser Tyr Glu Val Gln Ser Glu His Leu
1475 1480 1485
Pro His Arg Met Leu His Asn Trp Ala Leu Tyr Asn Ser Asp Leu
1490 1495 1500
Arg Phe Ile Ser Leu Glu Leu Leu Pro Met Lys Gln Cys Asp Asp
1505 1510 1515
Ile Asp Val Asn Ile Phe Gly Ser Gly Val Val Thr Asp Asp Asn
1520 1525 1530
Gly Ser Trp Ile Ser Leu Asn Asp Pro Asp Ser Gly Ser Gln Ser
1535 1540 1545
His Asp Pro Asp Gly Met Cys Ile Phe Leu Ser Gln Ile Lys Glu
1550 1555 1560
Trp Met Ile Glu Phe Gly Ser Asp Asp Ile Ile Ser Ile Ser Ile
1565 1570 1575
Arg Thr Asp Val Ala Trp Tyr Arg Leu Gly Lys Pro Ser Lys Leu
1580 1585 1590
Tyr Ala Pro Trp Trp Lys Pro Val Leu Lys Thr Ala Arg Val Gly
1595 1600 1605
Ile Ser Ile Leu Thr Phe Leu Arg Val Glu Ser Arg Val Ala Arg
1610 1615 1620
Leu Ser Phe Ala Asp Val Thr Lys Arg Leu Ser Gly Leu Gln Ala
1625 1630 1635
Asn Asp Lys Ala Tyr Ile Ser Ser Asp Pro Leu Ala Val Glu Arg
1640 1645 1650
Tyr Leu Val Val His Gly Gln Ile Ile Leu Gln Leu Phe Ala Val
1655 1660 1665
Tyr Pro Asp Asp Asn Val Lys Arg Cys Pro Phe Val Val Gly Leu
1670 1675 1680
Ala Ser Lys Leu Glu Asp Arg His His Thr Lys Trp Ile Ile Lys
1685 1690 1695
Lys Lys Lys Ile Ser Leu Lys Glu Leu Asn Leu Asn Pro Arg Ala
1700 1705 1710
Gly Met Ala Pro Val Ala Ser Lys Arg Lys Ala Met Gln Ala Thr
1715 1720 1725
Thr Thr Arg Leu Val Asn Arg Ile Trp Gly Glu Phe Tyr Ser Asn
1730 1735 1740
Tyr Ser Pro Glu Asp Pro Leu Gln Ala Thr Ala Ala Glu Asn Gly
1745 1750 1755
Glu Asp Glu Val Glu Glu Glu Gly Gly Asn Gly Glu Glu Glu Val
1760 1765 1770
Glu Glu Glu Gly Glu Asn Gly Leu Thr Glu Asp Thr Val Pro Glu
1775 1780 1785
Pro Val Glu Val Gln Lys Pro His Thr Pro Lys Lys Ile Arg Gly
1790 1795 1800
Ser Ser Gly Lys Arg Glu Ile Lys Trp Asp Gly Glu Ser Leu Gly
1805 1810 1815
Lys Thr Ser Ala Gly Glu Pro Leu Tyr Gln Gln Ala Leu Val Gly
1820 1825 1830
Gly Glu Met Val Ala Val Gly Gly Ala Val Thr Leu Glu Val Asp
1835 1840 1845
Asp Pro Asp Glu Met Pro Ala Ile Tyr Phe Val Glu Tyr Met Phe
1850 1855 1860
Glu Ser Thr Asp His Cys Lys Met Leu His Gly Arg Phe Leu Gln
1865 1870 1875
Arg Gly Ser Met Thr Val Leu Gly Asn Ala Ala Asn Glu Arg Glu
1880 1885 1890
Leu Phe Leu Thr Asn Glu Cys Met Thr Thr Gln Leu Lys Asp Ile
1895 1900 1905
Lys Gly Val Ala Ser Phe Glu Ile Arg Ser Arg Pro Trp Gly His
1910 1915 1920
Gln Tyr Arg Lys Lys Asn Ile Thr Ala Asp Lys Leu Asp Trp Ala
1925 1930 1935
Arg Ala Leu Glu Arg Lys Val Lys Asp Leu Pro Thr Glu Tyr Tyr
1940 1945 1950
Cys Lys Ser Leu Tyr Ser Pro Glu Arg Gly Gly Phe Phe Ser Leu
1955 1960 1965
Pro Leu Ser Asp Ile Gly Arg Ser Ser Gly Phe Cys Thr Ser Cys
1970 1975 1980
Lys Ile Arg Glu Asp Glu Glu Lys Arg Ser Thr Ile Lys Leu Asn
1985 1990 1995
Val Ser Lys Thr Gly Phe Phe Ile Asn Gly Ile Glu Tyr Ser Val
2000 2005 2010
Glu Asp Phe Val Tyr Val Asn Pro Asp Ser Ile Gly Gly Leu Lys
2015 2020 2025
Glu Gly Ser Lys Thr Ser Phe Lys Ser Gly Arg Asn Ile Gly Leu
2030 2035 2040
Arg Ala Tyr Val Val Cys Gln Leu Leu Glu Ile Val Pro Lys Glu
2045 2050 2055
Ser Arg Lys Ala Asp Leu Gly Ser Phe Asp Val Lys Val Arg Arg
2060 2065 2070
Phe Tyr Arg Pro Glu Asp Val Ser Ala Glu Lys Ala Tyr Ala Ser
2075 2080 2085
Asp Ile Gln Glu Leu Tyr Phe Ser Gln Asp Thr Val Val Leu Pro
2090 2095 2100
Pro Gly Ala Leu Glu Gly Lys Cys Glu Val Arg Lys Lys Ser Asp
2105 2110 2115
Met Pro Leu Ser Arg Glu Tyr Pro Ile Ser Asp His Ile Phe Phe
2120 2125 2130
Cys Asp Leu Phe Phe Asp Thr Ser Lys Gly Ser Leu Lys Gln Leu
2135 2140 2145
Pro Ala Asn Met Lys Pro Lys Phe Ser Thr Ile Lys Asp Asp Thr
2150 2155 2160
Leu Leu Arg Lys Lys Lys Gly Lys Gly Val Glu Ser Glu Ile Glu
2165 2170 2175
Ser Glu Ile Val Lys Pro Val Glu Pro Pro Lys Glu Ile Arg Leu
2180 2185 2190
Ala Thr Leu Asp Ile Phe Ala Gly Cys Gly Gly Leu Ser His Gly
2195 2200 2205
Leu Lys Lys Ala Gly Val Ser Asp Ala Lys Trp Ala Ile Glu Tyr
2210 2215 2220
Glu Glu Pro Ala Gly Gln Ala Phe Lys Gln Asn His Pro Glu Ser
2225 2230 2235
Thr Val Phe Val Asp Asn Cys Asn Val Ile Leu Arg Ala Ile Met
2240 2245 2250
Glu Lys Gly Gly Asp Gln Asp Asp Cys Val Ser Thr Thr Glu Ala
2255 2260 2265
Asn Glu Leu Ala Ala Lys Leu Thr Glu Glu Gln Lys Ser Thr Leu
2270 2275 2280
Pro Leu Pro Gly Gln Val Asp Phe Ile Asn Gly Gly Pro Pro Cys
2285 2290 2295
Gln Gly Phe Ser Gly Met Asn Arg Phe Asn Gln Ser Ser Trp Ser
2300 2305 2310
Lys Val Gln Cys Glu Met Ile Leu Ala Phe Leu Ser Phe Ala Asp
2315 2320 2325
Tyr Phe Arg Pro Arg Tyr Phe Leu Leu Glu Asn Val Arg Thr Phe
2330 2335 2340
Val Ser Phe Asn Lys Gly Gln Thr Phe Gln Leu Thr Leu Ala Ser
2345 2350 2355
Leu Leu Glu Met Gly Tyr Gln Val Arg Phe Gly Ile Leu Glu Ala
2360 2365 2370
Gly Ala Tyr Gly Val Ser Gln Ser Arg Lys Arg Ala Phe Ile Trp
2375 2380 2385
Ala Ala Ala Pro Glu Glu Val Leu Pro Glu Trp Pro Glu Pro Met
2390 2395 2400
His Val Phe Gly Val Pro Lys Leu Lys Ile Ser Leu Ser Gln Gly
2405 2410 2415
Leu His Tyr Ala Ala Val Arg Ser Thr Ala Leu Gly Ala Pro Phe
2420 2425 2430
Arg Pro Ile Thr Val Arg Asp Thr Ile Gly Asp Leu Pro Ser Val
2435 2440 2445
Glu Asn Gly Asp Ser Arg Thr Asn Lys Glu Tyr Lys Glu Val Ala
2450 2455 2460
Val Ser Trp Phe Gln Lys Glu Ile Arg Gly Asn Thr Ile Ala Leu
2465 2470 2475
Thr Asp His Ile Cys Lys Ala Met Asn Glu Leu Asn Leu Ile Arg
2480 2485 2490
Cys Lys Leu Ile Pro Thr Arg Pro Gly Ala Asp Trp His Asp Leu
2495 2500 2505
Pro Lys Arg Lys Val Thr Leu Ser Asp Gly Arg Val Glu Glu Met
2510 2515 2520
Ile Pro Phe Cys Leu Pro Asn Thr Ala Glu Arg His Asn Gly Trp
2525 2530 2535
Lys Gly Leu Tyr Gly Arg Leu Asp Trp Gln Gly Asn Phe Pro Thr
2540 2545 2550
Ser Val Thr Asp Pro Gln Pro Met Gly Lys Val Gly Met Cys Phe
2555 2560 2565
His Pro Glu Gln His Arg Ile Leu Thr Val Arg Glu Cys Ala Arg
2570 2575 2580
Ser Gln Gly Phe Pro Asp Ser Tyr Glu Phe Ala Gly Asn Ile Asn
2585 2590 2595
His Lys His Arg Gln Ile Gly Asn Ala Val Pro Pro Pro Leu Ala
2600 2605 2610
Phe Ala Leu Gly Arg Lys Leu Lys Glu Ala Leu His Leu Lys Lys
2615 2620 2625
Ser Pro Gln His Gln Pro
2630
<210> 29
<211> 1113
<212> PRT
<213> RB-vTALE-AtSRDX氨基酸序列
<400> 29
Met Asp Pro Ile Arg Ser Arg Thr Pro Ser Pro Ala Arg Glu Leu Leu
1 5 10 15
Pro Gly Pro Gln Pro Asp Arg Val Gln Pro Thr Ala Asp Arg Gly Gly
20 25 30
Ala Pro Pro Ala Gly Gly Pro Leu Asp Gly Leu Pro Ala Arg Arg Thr
35 40 45
Met Ser Arg Thr Arg Leu Pro Ser Pro Pro Ala Pro Ser Pro Ala Phe
50 55 60
Ser Ala Gly Ser Phe Asn Asp Leu Leu Arg Gln Phe Asp Pro Ser Leu
65 70 75 80
Leu Asp Thr Ser Leu Leu Asp Ser Met Pro Ala Val Gly Thr Pro His
85 90 95
Thr Ala Ala Ala Pro Ala Glu Trp Asp Glu Val Gln Ser Gly Leu Arg
100 105 110
Ala Ala Asp Asp Pro Pro Pro Thr Val Arg Val Ala Val Thr Ala Ala
115 120 125
Arg Pro Pro Arg Ala Lys Pro Ala Pro Arg Arg Arg Ala Ala Gln Pro
130 135 140
Ser Asp Ala Ser Pro Ala Ala Gln Val Asp Leu Arg Thr Leu Gly Tyr
145 150 155 160
Ser Gln Gln Gln Gln Glu Lys Ile Lys Ser Lys Val Arg Ser Thr Val
165 170 175
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
180 185 190
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
195 200 205
Lys Tyr Gln His Ile Ile Thr Ala Leu Pro Glu Ala Thr His Glu Asp
210 215 220
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
225 230 235 240
Leu Leu Thr Lys Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
245 250 255
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
260 265 270
Glu Ala Val His Ala Ser Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
275 280 285
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys
290 295 300
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
305 310 315 320
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly
325 330 335
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
340 345 350
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn
355 360 365
Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
370 375 380
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
385 390 395 400
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
405 410 415
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
420 425 430
Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
435 440 445
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
450 455 460
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
465 470 475 480
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
485 490 495
Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu
500 505 510
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
515 520 525
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
530 535 540
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
545 550 555 560
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
565 570 575
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
580 585 590
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly
595 600 605
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
610 615 620
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His
625 630 635 640
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
645 650 655
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
660 665 670
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
675 680 685
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
690 695 700
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
705 710 715 720
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
725 730 735
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
740 745 750
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
755 760 765
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
770 775 780
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
785 790 795 800
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
805 810 815
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
820 825 830
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
835 840 845
Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala
850 855 860
Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly
865 870 875 880
Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Pro His Ala Pro
885 890 895
Glu Leu Ile Arg Arg Ile Asn Arg Arg Ile Pro Glu Arg Thr Ser His
900 905 910
Arg Val Ala Asp Leu Ala His Val Val Arg Val Leu Gly Phe Phe Gln
915 920 925
Ser His Ser His Pro Ala Gln Ala Phe Asp Asp Ala Met Thr Gln Phe
930 935 940
Gly Met Ser Arg His Gly Leu Val Gln Leu Phe Arg Arg Val Gly Val
945 950 955 960
Thr Glu Phe Glu Ala Arg Cys Gly Thr Leu Pro Pro Ala Ser Gln Arg
965 970 975
Trp Asp Arg Ile Leu Gln Ala Ser Gly Met Lys Arg Ala Lys Pro Ser
980 985 990
Pro Thr Ser Ala Gln Thr Pro Asp Gln Ala Ser Leu His Ala Phe Ala
995 1000 1005
Asp Ser Leu Glu Arg Asp Leu Asp Ala Pro Ser Pro Met His Glu
1010 1015 1020
Gly Asp Gln Thr Arg Ala Ser Ser Arg Lys Arg Ser Arg Ser Asp
1025 1030 1035
Arg Ala Val Thr Gly Pro Ser Thr Gln Gln Ser Phe Glu Val Arg
1040 1045 1050
Val Pro Glu Gln Arg Asp Ala Leu His Leu Pro Leu Ser Trp Arg
1055 1060 1065
Val Lys Arg Pro Arg Thr Arg Ile Gly Gly Gly Leu Pro Asp Pro
1070 1075 1080
Gly Thr Pro Ile Ala Ala Asp Leu Ala Ala Ser Ser Thr Val Met
1085 1090 1095
Trp Ser Ser Leu Asp Leu Asp Leu Glu Leu Arg Leu Gly Phe Ala
1100 1105 1110
<210> 30
<211> 1751
<212> PRT
<213> RB-vTALE-AtSUVH2氨基酸序列
<400> 30
Met Asp Pro Ile Arg Ser Arg Thr Pro Ser Pro Ala Arg Glu Leu Leu
1 5 10 15
Pro Gly Pro Gln Pro Asp Arg Val Gln Pro Thr Ala Asp Arg Gly Gly
20 25 30
Ala Pro Pro Ala Gly Gly Pro Leu Asp Gly Leu Pro Ala Arg Arg Thr
35 40 45
Met Ser Arg Thr Arg Leu Pro Ser Pro Pro Ala Pro Ser Pro Ala Phe
50 55 60
Ser Ala Gly Ser Phe Asn Asp Leu Leu Arg Gln Phe Asp Pro Ser Leu
65 70 75 80
Leu Asp Thr Ser Leu Leu Asp Ser Met Pro Ala Val Gly Thr Pro His
85 90 95
Thr Ala Ala Ala Pro Ala Glu Trp Asp Glu Val Gln Ser Gly Leu Arg
100 105 110
Ala Ala Asp Asp Pro Pro Pro Thr Val Arg Val Ala Val Thr Ala Ala
115 120 125
Arg Pro Pro Arg Ala Lys Pro Ala Pro Arg Arg Arg Ala Ala Gln Pro
130 135 140
Ser Asp Ala Ser Pro Ala Ala Gln Val Asp Leu Arg Thr Leu Gly Tyr
145 150 155 160
Ser Gln Gln Gln Gln Glu Lys Ile Lys Ser Lys Val Arg Ser Thr Val
165 170 175
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
180 185 190
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
195 200 205
Lys Tyr Gln His Ile Ile Thr Ala Leu Pro Glu Ala Thr His Glu Asp
210 215 220
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
225 230 235 240
Leu Leu Thr Lys Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
245 250 255
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
260 265 270
Glu Ala Val His Ala Ser Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
275 280 285
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys
290 295 300
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
305 310 315 320
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly
325 330 335
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
340 345 350
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn
355 360 365
Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
370 375 380
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
385 390 395 400
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
405 410 415
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
420 425 430
Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
435 440 445
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
450 455 460
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
465 470 475 480
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
485 490 495
Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu
500 505 510
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
515 520 525
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
530 535 540
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
545 550 555 560
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
565 570 575
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
580 585 590
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly
595 600 605
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
610 615 620
Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His
625 630 635 640
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
645 650 655
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
660 665 670
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
675 680 685
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
690 695 700
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
705 710 715 720
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
725 730 735
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
740 745 750
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp
755 760 765
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
770 775 780
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
785 790 795 800
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
805 810 815
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
820 825 830
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
835 840 845
Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala
850 855 860
Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly
865 870 875 880
Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Pro His Ala Pro
885 890 895
Glu Leu Ile Arg Arg Ile Asn Arg Arg Ile Pro Glu Arg Thr Ser His
900 905 910
Arg Val Ala Asp Leu Ala His Val Val Arg Val Leu Gly Phe Phe Gln
915 920 925
Ser His Ser His Pro Ala Gln Ala Phe Asp Asp Ala Met Thr Gln Phe
930 935 940
Gly Met Ser Arg His Gly Leu Val Gln Leu Phe Arg Arg Val Gly Val
945 950 955 960
Thr Glu Phe Glu Ala Arg Cys Gly Thr Leu Pro Pro Ala Ser Gln Arg
965 970 975
Trp Asp Arg Ile Leu Gln Ala Ser Gly Met Lys Arg Ala Lys Pro Ser
980 985 990
Pro Thr Ser Ala Gln Thr Pro Asp Gln Ala Ser Leu His Ala Phe Ala
995 1000 1005
Asp Ser Leu Glu Arg Asp Leu Asp Ala Pro Ser Pro Met His Glu
1010 1015 1020
Gly Asp Gln Thr Arg Ala Ser Ser Arg Lys Arg Ser Arg Ser Asp
1025 1030 1035
Arg Ala Val Thr Gly Pro Ser Thr Gln Gln Ser Phe Glu Val Arg
1040 1045 1050
Val Pro Glu Gln Arg Asp Ala Leu His Leu Pro Leu Ser Trp Arg
1055 1060 1065
Val Lys Arg Pro Arg Thr Arg Ile Gly Gly Gly Leu Pro Asp Pro
1070 1075 1080
Gly Thr Pro Ile Ala Ala Asp Leu Ala Ala Ser Ser Thr Val Met
1085 1090 1095
Trp Ser Ser Ser Thr Leu Leu Pro Phe Pro Asp Leu Asn Leu Met
1100 1105 1110
Pro Asp Ser Gln Ser Ser Thr Ala Gly Thr Thr Ala Gly Asp Thr
1115 1120 1125
Val Val Thr Gly Lys Leu Glu Val Lys Ser Glu Pro Ile Glu Glu
1130 1135 1140
Trp Gln Thr Pro Pro Ser Ser Thr Ser Asp Gln Ser Ala Asn Thr
1145 1150 1155
Asp Leu Ile Ala Glu Phe Ile Arg Ile Ser Glu Leu Phe Arg Ser
1160 1165 1170
Ala Phe Lys Pro Leu Gln Val Lys Gly Leu Asp Gly Val Ser Val
1175 1180 1185
Tyr Gly Leu Asp Ser Gly Ala Ile Val Ala Val Pro Glu Lys Glu
1190 1195 1200
Asn Arg Glu Leu Ile Glu Pro Pro Pro Gly Phe Lys Asp Asn Arg
1205 1210 1215
Val Ser Thr Val Val Val Ser Pro Lys Phe Glu Arg Pro Arg Glu
1220 1225 1230
Leu Ala Arg Ile Ala Ile Leu Gly His Glu Gln Arg Lys Glu Leu
1235 1240 1245
Arg Gln Val Met Lys Arg Thr Arg Met Thr Tyr Glu Ser Leu Arg
1250 1255 1260
Ile His Leu Met Ala Glu Ser Met Lys Asn His Val Leu Gly Gln
1265 1270 1275
Gly Arg Arg Arg Arg Ser Asp Met Ala Ala Ala Tyr Ile Met Arg
1280 1285 1290
Asp Arg Gly Leu Trp Leu Asn Tyr Asp Lys His Ile Val Gly Pro
1295 1300 1305
Val Thr Gly Val Glu Val Gly Asp Ile Phe Phe Tyr Arg Met Glu
1310 1315 1320
Leu Cys Val Leu Gly Leu His Gly Gln Thr Gln Ala Gly Ile Asp
1325 1330 1335
Cys Leu Thr Ala Glu Arg Ser Ala Thr Gly Glu Pro Ile Ala Thr
1340 1345 1350
Ser Ile Val Val Ser Gly Gly Tyr Glu Asp Asp Glu Asp Thr Gly
1355 1360 1365
Asp Val Leu Val Tyr Thr Gly His Gly Gly Gln Asp His Gln His
1370 1375 1380
Lys Gln Cys Asp Asn Gln Arg Leu Val Gly Gly Asn Leu Gly Met
1385 1390 1395
Glu Arg Ser Met His Tyr Gly Ile Glu Val Arg Val Ile Arg Gly
1400 1405 1410
Ile Lys Tyr Glu Asn Ser Ile Ser Ser Lys Val Tyr Val Tyr Asp
1415 1420 1425
Gly Leu Tyr Lys Ile Val Asp Trp Trp Phe Ala Val Gly Lys Ser
1430 1435 1440
Gly Phe Gly Val Phe Lys Phe Arg Leu Val Arg Ile Glu Gly Gln
1445 1450 1455
Pro Met Met Gly Ser Ala Val Met Arg Phe Ala Gln Thr Leu Arg
1460 1465 1470
Asn Lys Pro Ser Met Val Arg Pro Thr Gly Tyr Val Ser Phe Asp
1475 1480 1485
Leu Ser Asn Lys Lys Glu Asn Val Pro Val Phe Leu Tyr Asn Asp
1490 1495 1500
Val Asp Gly Asp Gln Glu Pro Arg His Tyr Glu Tyr Ile Ala Lys
1505 1510 1515
Ala Val Phe Pro Pro Gly Ile Phe Gly Gln Gly Gly Ile Ser Arg
1520 1525 1530
Thr Gly Cys Glu Cys Lys Leu Ser Cys Thr Asp Asp Cys Leu Cys
1535 1540 1545
Ala Arg Lys Asn Gly Gly Glu Phe Ala Tyr Asp Asp Asn Gly His
1550 1555 1560
Leu Leu Lys Gly Lys His Val Val Phe Glu Cys Gly Glu Phe Cys
1565 1570 1575
Thr Cys Gly Pro Ser Cys Lys Ser Arg Val Thr Gln Lys Gly Leu
1580 1585 1590
Arg Asn Arg Leu Glu Val Phe Arg Ser Lys Glu Thr Gly Trp Gly
1595 1600 1605
Val Arg Thr Leu Asp Leu Ile Glu Ala Gly Ala Phe Ile Cys Glu
1610 1615 1620
Tyr Ala Gly Val Val Val Thr Arg Leu Gln Ala Glu Ile Leu Ser
1625 1630 1635
Met Asn Gly Asp Val Met Val Tyr Pro Gly Arg Phe Thr Asp Gln
1640 1645 1650
Trp Arg Asn Trp Gly Asp Leu Ser Gln Val Tyr Pro Asp Phe Val
1655 1660 1665
Arg Pro Asn Tyr Pro Ser Leu Pro Pro Leu Asp Phe Ser Met Asp
1670 1675 1680
Val Ser Arg Met Arg Asn Val Ala Cys Tyr Ile Ser His Ser Lys
1685 1690 1695
Glu Pro Asn Val Met Val Gln Phe Val Leu His Asp His Asn His
1700 1705 1710
Leu Met Phe Pro Arg Val Met Leu Phe Ala Leu Glu Asn Ile Ser
1715 1720 1725
Pro Leu Ala Glu Leu Ser Leu Asp Tyr Gly Leu Ala Asp Glu Val
1730 1735 1740
Asn Gly Lys Leu Ala Ile Cys Asn
1745 1750

Claims (10)

1.一种改造的抗双生病毒的vTALE分子，其以植物病原细菌水稻黄单胞杆菌(Xanthomonas oryzae pv.oryzae)三型分泌系统效应因子AvrXa27基因序列为骨架序列，保留AvrXa27基因的N端和C端序列，然后对AvrXa27基因的DNA识别/结合结构域序列和功能输出结构域序列进行改造而获得，其中所述改造后的DNA识别/结合结构域序列靶向SEQ IDNO：1所示的IR-vTALE靶点序列或SEQ ID NO：2所示的RB-vTALE靶点序列，并且所述改造后的功能输出结构域序列选自由编码SEQ ID NOs：8-12所示的氨基酸序列的核苷酸序列组成的组中的任一种。
2.权利要求1所述的改造的抗双生病毒的vTALE分子，其中所述改造后的DNA识别/结合结构域序列靶向SEQ ID NO：1所示的IR-vTALE靶点序列。
3.权利要求1所述的改造的抗双生病毒的vTALE分子，其中所述改造后的DNA识别/结合结构域序列靶向SEQ ID NO：2所示的RB-vTALE靶点序列。
4.权利要求1所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列编码SEQ ID NO：8所示的AtKYP蛋白。
5.权利要求1所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列编码SEQ ID NO：9所示的AtHDAC19蛋白。
6.权利要求1所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列编码SEQ ID NO：10所示的AtMET1蛋白。
7.权利要求1所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列编码SEQ ID NO：11所示的AtSRDX蛋白。
8.权利要求1所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列编码SEQ ID NO：12所示的AtSUVH2蛋白。
9.权利要求1所述的改造的抗双生病毒的vTALE分子，其中所述改造后的功能输出结构域序列是选自由SEQ ID NOs：3-7所示的核苷酸序列组成的组的任一种核苷酸序列。
10.一种培育具有稳定的广谱双生病毒抗性的植物的方法，所述方法包括下述步骤：
(1)vTALE分子的构建：所述vTALE分子以植物病原细菌水稻黄单胞杆菌(Xanthomonasoryzae pv.oryzae)三型分泌系统效应因子AvrXa27基因序列为骨架序列，保留AvrXa27基因的N端和C端序列，然后对AvrXa27基因的DNA识别/结合结构域序列和功能输出结构域序列进行改造而获得，其中所述改造后的DNA识别/结合结构域序列靶向SEQ ID NO：1所示的IR-vTALE靶点序列或SEQ ID NO：2所示的RB-vTALE靶点序列；并且所述改造后的功能输出结构域序列选自由编码SEQ ID NOs：8-12所示的氨基酸序列的核苷酸序列组成的组中的任一种；
(2)植物转化：将步骤(1)构建的vTALE分子，转化到所述植物中进行稳定表达。
CN201611222101.9A 2016-12-26 2016-12-26 一种提高植物对双生病毒抗性的方法 Pending CN108277228A (zh)

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