CN101128590A - 产量增加的植物及其制备方法 - Google Patents
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Abstract
本发明涉及相对于在非胁迫生长条件下生长的对应野生型植物,在相当条件下生长的植物中增加植物产量的方法,该方法包括在植物细胞的细胞溶胶中优先增加I型DnaJ样多肽或其同源物的活性。此类方法中的一种包括在植物、植物部分或植物细胞中引入和/或表达I型DnaJ样核酸或其变体。本发明还涉及在非胁迫条件下生长已经在其中引入和/或表达了I型DnaJ样核酸或其变体的转基因植物,该植物相对于在相当条件下生长的对应野生型植物具有增加的植物产量。本发明还涉及可用在本发明方法中的构建体。
Description
本发明通常涉及分子生物学领域,并且涉及相对于在非胁迫条件下生长的对应野生型植物,在相当条件下生长的植物中增加植物产量的方法。更具体的,本发明涉及在非胁迫条件下生长的植物中增加植物产量的方法,包括在植物细胞溶胶中优先增加I型DnaJ样多肽或其同源物的活性。本发明还涉及在细胞溶胶中优先增加了I型DnaJ样多肽或其同源物的活性的植物,当该植物在非胁迫条件下生长时,相对于在相当条件下生长的对应的野生型植物,具有增加的产量。本发明还提供了可用于本发明方法中的构建体。
不断增长的世界人口和农业用地面积供应的缩小激起了对增加农业效率的农业研究。作物和园艺改良的常规手段利用选育技术来鉴定具有期望特性的植物。然而,这种选育技术具有若干缺点,即这些技术一般是劳动密集型的,而且产生通常含有异质遗传成分的植物,所述植物并非总是产生自亲本植物传递过来的期望性状。分子生物学的发展已经允许人类操作动物和植物的种质。植物的遗传工程需要分离和操作遗传物质(一般是DNA或RNA的形式),随后把该遗传物质引入到植物中。此种技术能够提供具有多种改良的经济学、农艺学或园艺性状的植物。产出了具有特殊经济利益的性状。产量通常定义为作物产生的可衡量经济价值。这可以从数量和/或质量方面定义。产量直接取决于一些因素,例如,器官的数量和尺寸,植物结构(例如,枝条的数量),种子产量等等。根的发育、营养吸收和胁迫耐受性也是确定产量的重要因素。优化上述因素之一可以因此提高作物产量。另外,植物种子是人和动物营养的重要来源。作物如玉米、稻、小麦、卡诺拉油菜和大豆为超过半数的总人类的热量摄取,不管是通过种子本身的直接消耗还是通过加工的种子饲养的肉类产品的消耗。它们还是糖、油和用在工业加工中的许多类代谢物的来源。种子包含在萌发后新枝条和根的来源——胚胎,以及在萌发和幼苗的早期生长过程中胚胎生长养料的来源—胚乳。种子发育包括许多基因,并且需要将来自根、叶和茎的代谢物转移到生长的种子中。特别的,胚乳吸收糖类聚合物、油和蛋白质的代谢前体并且将它们合成为贮藏大分子以灌浆谷粒。无论是通过增加的收获指数、增加的千粒重、种子数量、种子生物量、种子发育、种子饱满或任何其他种子相关性状的增加植物种子产量的能力都在农业并且甚至是许多非农业用途(例如在物质如药物、抗体或疫苗的生物技术制备中)中具有许多应用。
已经发现在植物细胞的细胞溶胶中优先增加I型DnaJ样多肽的活性使得在非胁迫条件下生长的植物相对于在相当条件下生长的对应野生型植物具有增加的产量。
DnaJ是Hsp40(热休克蛋白40)家族的分子辅陪伴分子。Hsp40与陪伴分子热休克蛋白70(Hsp70,也称作DnaK)和辅陪伴分子核苷酸交换因子GrpE协同作用来促进细胞蛋白质代谢的不同方面,包括核糖体装配、蛋白质转运、蛋白质折叠和蛋白质解折叠、多肽集合的阻抑和细胞信号传导(Walid(2001)Curr Protein Peptide Sci 2:227-244)。DnaJ刺激Hsp70水解ATP,这是底物与Hsp70稳定结合的关键步骤。另外,由于已经表明DnaJ在体外翻译系统中结合到新生链上并且阻止变性多肽的聚集,DnaJ本身也具有分子陪伴分子功能(Laufen等人(2001)Proc Natl Acad Sci美国96:5452-5457)。已经在多种生物(在原核生物和真核生物中)中以及在多种细胞区室(如细胞溶胶、线粒体、过氧化物酶体、乙醛酸循环体、内质网和叶绿体基质)中鉴定了DnaJ家族的成员。在一种生物中,多个Hsp40可以与单个Hsp70相互作用以生成Hsp70::Hsp40对,这促进了在细胞蛋白质代谢中的多种反应。
所有的DnaJ蛋白质是通过所谓的“J”结构域的存在并且通过在J结构域中间高度保守的HPD三肽的存在来定义的,所述“J”结构域由大约70个氨基酸组成,通常位于该蛋白质的氨基末端(InterPro参考IPR001623;Zdobnov等人,(2002)18(8):1149-50);由4个α螺旋组成的“J”结构域与Hsp70蛋白质相互作用。在拟南芥(Arabidopsis thaliana)基因组中,已经鉴定了至少89种包含J-结构域的蛋白质(Miernyk(2001)Cell Stress&Chaperones)。目前已经鉴定了18种Hsp70蛋白质。
DnaJ蛋白质被进一步分成I型、II型和III型。
I型DnaJ结构域蛋白质(或DnaJ蛋白质)(目前在拟南芥属(Arabidopsis)中存在至少8种;Miernyk(2001)Cell Stress&Chaperone 6(3):209-218)包含(从氨基末端到羧基末端)在原始型DnaJ蛋白质(如在大肠杆菌(Eschriehia coli)中首先鉴定的)中鉴定的结构域:
1)大约30个氨基酸残基的G/F结构域区域,其富含甘氨酸(G)和苯丙氨酸(F),其用来调节靶标多肽特异性;
2)包含4个重复CXXCXGXG的富含Cys的锌指结构域,其中X代表了带电或极性残基;这4个重复片段成对作用来形成锌结合结构域I和II(InterPro参考IPR001305;Linke等人(2003)J Biol Chem 278(45):44457-44466);认为锌指结构域介导直接的蛋白质:蛋白质相互作用并且更特异性的结合待递送到Hsp70的非天然多肽;
3)羧基末端结构域(CTD;InterPro参考IPR002939)。
II型DnaJ结构域蛋白质(目前在拟南芥属中存在至少35种)包含位于蛋白质氨基末端的J结构域、G/F结构域或锌指结构域,以及CTD。III型DnaJ结构域蛋白质(目前在拟南芥属中存在至少45种)仅包含J结构域,其位于蛋白质中的任何位置。
在其天然形式中(可以是可溶的或膜结合的形式),DnaJ蛋白质可以靶向到多个亚细胞区室。在植物中此类亚细胞区室的实例包括线粒体、叶绿体、过氧化物酶体、核、细胞质和分泌途径。通常位于核编码的DnaJ蛋白质的氨基末端的信号序列和转运肽可用于将这些蛋白质靶向到特异性的亚细胞区室中。
在特异性环境下DnaJ蛋白质所靶向的细胞膜实例包括线粒体外膜、叶绿体外膜、过氧化物酶体膜、核膜、内质网(ER)和细胞膜本身(Miernyk(2001)Cell Stress&Chaperone 6(3):209-218)。DnaJ蛋白质的一种类型的膜结合发生在蛋白质的翻译后修饰后(即在异戊二烯化后)。类异戊二烯基团连接到位于蛋白质羧基末端末的法尼基化CaaX基序(其中C是Cys,a是脂族氨基酸残基并且X是任何氨基酸)的半胱氨酸。已经表明此法尼基化导致更高的生物学活性和DnaJ蛋白质的膜结合,特别是在升高的温度下(Zhu J-K等人,(1993)The Plant Cell 5:341-9)。
已经暗示DnaJ蛋白质对赋予植物热胁迫耐受性中发挥作用(与HSP70一起)。同时DnaJ在热胁迫的植物中起到保护性作用,不明确在非热胁迫的植物中增加DnaJ水平和/或活性是否存在其他任何的优势。
因此令人惊讶的发现I型DnaJ样多肽可以在非胁迫生长条件下使用,以使得植物相对于在相当条件下生长的对应野生型植物的产量具有增加的产量。
因此,根据本发明的一个实施方案,提供了在非胁迫条件下生长的植物中增加植物产量的方法,其包括在植物细胞的细胞溶胶中优先增加I型DnaJ样多肽或其同源物的活性,该I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序。
术语“细胞溶胶”是指用于本发明方法中的DnaJ蛋白质的亚细胞位置。DnaJ蛋白质与线粒体、叶绿体、过氧化物酶体、核、ER的外表面或与细胞膜的瞬时或长久结合涵盖在术语细胞溶胶中。
蛋白质的“羧基末端”可以由本领域的技术人员容易的鉴定出来。
本文提及的“对应野生型植物”意指任何适当的对照植物,其选择是本领域技术人员能力范围内的,并且可以包括例如对应的野生型植物或无目的基因的对应植物。本文所用的“对照植物”不仅是指整株植物,还是指植物部分,包括种子和种子部分。
本文提及的“非胁迫(生长)条件”意指在正常生长条件下生长/培育在其生活周期(从种子到成熟植物再到种子)任何阶段的植物,所述正常生长条件包括每株植物都会遇到的每天的中等胁迫,但是不包括严重胁迫。严重胁迫的实例包括热胁迫,其发生是本领域熟知的,并且取决于多种因素,如植物生长的区域并且取决于植物自身。
有利的,实施本发明方法产生了具有增加的产量,特别是增加的种子产量的植物。
本文所定义的术语“增加的产量”意指分别相对于对应的野生型植物,如下任何一个或多个方面的增加:(i)植物的一个或多个部分增加的生物量(重量),特别是地上(可收获)部分、增加的根生物量或任何其他可收获部分的增加的生物量;(ii)增加的种子产量,其包括种子的生物量(种子重量)增加,并且其可以是每株植株或单粒种子基础上的种子重量增加;(iii)增加的(饱满)种子数;(iv)增加的种子饱满率(其是饱满种子数除以总种子数并且乘以100的数量);(v)增加的种子尺寸,其还可以影响种子组成;(vi)增加的种子体积,这也可影响种子的组成;(vii)增加的收获指数,其表示为可收获部分(例如种子)的产量与总生物量的比例;和(viii)增加的千粒重(TKW),其从所计的饱满种子数和它们的总重量推论出来。增加的TKW可源于增加的种子尺寸和/或种子重量。
以谷类为例,产量增加可以表现为下列一种或多种:每公顷或英亩植物数量增加、每株植物穗数量的增加、畦数、每畦种子数、粒重、千粒重、穗的长度/直径的增加等。以稻为例,产量增加可以表现为下列一种或多种的增加:每公顷或英亩植物数量、每株植物的圆锥花序数量、每个圆锥花序的小穗数量、每个圆锥花序花的数量,种子饱满率的增加、千粒重的增加等。产量的增加还可产生改变的构造或可以因为改变的构造而发生。
根据优选的特征,实施本发明方法产生具有增加的种子产量的植物。因此,根据本发明,提供了在非胁迫条件下生长的植物中增加植物种子产量的方法,该方法包括在植物细胞的细胞溶胶中优先增加I型DnaJ样多肽或其同源物的活性,该I型DnaJ样多肽在其羧基末端包含CaaX基序。
另外优选的,增加的种子产量表现为相对于对应野生型植物的收获指数的增加。因此,根据本发明,提供了在非胁迫条件下生长的植物中增加收获指数的方法,该方法包括在植物细胞的细胞溶胶中优先增加I型DnaJ样多肽或其同源物的活性,该I型DnaJ样多肽在其羧基末端包含CaaX基序。
由于根据本发明的转基因植物具有增加的产量,因此,相对于在其生活周期对应阶段的相应野生型植物的生长速度,这些植物很可能表现出增加的生长速度(至少是在其生活周期的一部分)。生长速度增加可以是对植物的一个或多个部分(包括种子)特异性的,或者可以基本遍布整株植物。具有增加的生长速度的植物甚至可以表现出早开花。生长速度的增加可以发生在植物生活周期的一个或多个阶段或者基本遍布整个植物生活周期期间。在植物生活周期的早期阶段增加的生长速度反映了增强的活力。生长速度的增加可以改变植物的收获周期,这使得植物可以比可能的情形更晚播种和/或更早收获。如果生长速度充分增加,可以允许更多播种相同植物物种的种子(例如在播种和收获稻植物后,接着播种和收获更多的稻植物,所有这些都在一个常规生长期内)。类似的,如果生长速度充分增加,可以允许更多播种不同植物物种的种子(例如在播种和收获稻植物后,例如接着播种和任选收获大豆、马铃薯或任何其他适当的植物作物)。在一些植物的情形下,从同一根状茎收获多次也是可能的。改变植物的收获周期可以使得每英亩每年生物量的产出增加(由于任何特定植物可以生长和收获次数(指在一年中)的增加)。生长速度的增加还使得转基因植物可以比它们的野生型对应物在更广的地域栽培,这是因为生长作物的区域性限制通常决定于种植时(初期)或收获时(晚期)的不利环境条件。如果收获周期缩短,那么可以避免此类不利条件。可以通过得自生长曲线的多个参数来确定生长速度,此类参数可以是:T-Mid(植物达到其最大尺寸的50%时所用的时间)以及T-90(植物达到其最大尺寸的90%时所用的时间)等。
本发明方法的实施提供了具有增加的生长速度的植物。因此,根据本发明,提供了相对于在非胁迫条件下生长的野生型植物的生长速度,增加在相当条件下生长的植物的生长速度的方法,该方法包括在植物细胞的细胞溶胶中优先增加I型DnaJ样多肽或其同源物的活性,该I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序。
在任何植物中可以有利的修饰上述生长特性。
本文所用的术语“植物”包括整株植物、植物的祖代和后代以及植物部分(包括种子、枝条、茎、叶、根(包括块茎)、花以及组织和器官,其中前述每种包括目的基因/核酸。术语“植物”还包括植物细胞、悬浮培养物、愈伤组织、胚、分生组织区、配子体、孢子体、花粉和小孢子,同样的,其中前述每种包含目的基因/核酸。
尤其可用于本发明方法中的植物包括属于植物界(Viridiplantae)超家族的所有植物,尤其是单子叶植物和双子叶植物,包括选自下列的饲料或饲料豆科植物、观赏植物、粮食作物、乔木或灌木:金合欢属物种(Acaciaspp.)、槭树属物种(Acer spp.)、猕猴桃属物种(Actinidia spp.)、七叶树属物种(Aesculus spp.)、新西兰贝壳杉(Agathis australis)、Albizia amara、三色桫椤(Alsophila tricolor)、须芒草属物种(Andropogon spp.)、落花生属物种(Arachis spp)、槟榔(Areca catechu)、Astelia fragrans、黄芪(Astragaluscicer)、Baikiaea plurijuga、桦木属物种(Betula spp.)、芸苔属物种(Brassicaspp.)、木榄(Bruguiera gymnorrhiza)、Burkea africana、紫铆(Buteafrondosa)、Cadaba farinosa、朱缨花属物种(Calliandra spp)、茶(Camelliasinensis)、美人蕉(Canna indica)、辣椒属物种(Capsicum spp.)、决明属物种(Cassia spp.)、距瓣豆(Centroema pubescens)、木瓜属物种(Chaenomelesspp.)、肉桂(Cinnamomum cassia)、小果咖啡(Coffea arabica)、Colophospermum mopane、变异小冠花(Coronillia varia)、枸子(Cotoneaster serotina)、山楂属物种(Crataegus spp.)、香瓜属物种(Cucumisspp.)、柏木属物种(Cupressus spp.)、Cyathea dealbata、木梨(Cydoniaoblonga)、圆球柳杉(Cryptomeria japonica)、香茅属物种(Cymbopogonspp.)、Cynthea dealbata、木梨(Cydonia oblonga)、Dalbergia monetaria、大叶骨碎补(Davallia divaricata)、山马蝗属物种(Desmodium spp.)、迪卡兰(Dicksonia squarosa)、Diheteropogon amplectens、Dioclea spp、镰扁豆属物种(Dolichos spp.)、Dorycnium rectum、锥穗稗(Echinochloapyramidalis)、Ehrartia spp.、穇子(Eleusine coracana)、Eragrestis spp.、刺桐属物种(Erythrina spp.)、桉属物种(Eucalyptus spp.)、Eucleaschimperi、金茅(Eulalia villosa)、荞麦属物种(Fagopyrum spp.)、费约罗(Feijoa sellowiana)、草雷属物种(Fragaria spp.)、千斤拔属物种(Flemingiaspp)、Freycinetia banksii、Geranium thunbergii、银杏(Ginkgo biloba)、Glycine javanica、Gliricidia spp、陆地棉(Gossypium hirsutum)、银桦属物种(Grevillea spp.)、Guibourtia coleosperma、岩黄芪属物种(Hedysarumspp.)、牛鞭草(Hemarthia altissima)、扭黄茅(Heteropogon contortus)、大麦(Hordeum vulgare)、Hyparrhenia rufa、小连翅(Hypericum erectum)、Hyperthelia dissoluta、白花庭蓝(Indigo incarnata)、鸢尾属物种(Iris spp.)、Leptarrhena pyrolifolia、胡枝子属物种(Lespediza spp.)、Lettuca spp.、Leucaena leucocephala、Loudetia simplex、Lotonus bainesii、百脉根属物种(Lotus spp.)、硬皮豆(Macrotyloma axillare)、苹果属物种(Malus spp.)、Manihot esculenta、紫苜蓿(Medicago sativa)、水杉(Metasequoiaglyptostroboides)、大蕉(Musa sapientum)、烟草属物种(Nicotianum spp.)、驴食草属物种(Onobrychis spp.)、Ornithopus spp.、稻属物种(Oryza spp.)、非洲双翼豆(Peltophorum africanum)、狼尾草属物种(Pennisetum spp.)、鳄梨(Persea gratissima)、碧冬茄属物种(Petunia spp.)、菜豆属物种(Phaseolus spp.)、槟榔竹(Phoenix canariensis)、Phormium cookianum、石楠属物种(Photinia spp.)、白云杉(Picea glauca)、松属物种(Pinus spp.)、豌豆(Pisum sativum)、新西兰罗汉松(Podocarpus totara)、Pogonarthriafieckii、Pogonarthria squarrosa、杨属物种(Populus spp.)、牧豆树(Prosopiscineraria)、花旗松(Pseudotsuga menziesii)、Pterolobium stellatum、西洋梨(Pyrus communis)、栎属物种(Quercus spp.)、Rhaphiolepsis umbellata、美味棒花棕(Rhopalostylis sapida)、Rhus natalensis、欧洲醋粟(Ribesgrossularia)、茶子属物种(Ribes spp.)、洋槐(Robinia pseudoacacia)、蔷薇属物种(Rosa spp.)、悬钩子属物种(Rubus spp.)、柳属物种(Salix spp.)、Schyzachyrium sanguineum、金松(Sciadopitys verticillata)、北美红杉(Sequoia sempervirens)、巨杉(Sequoiadendron giganteum)、两色蜀黍(Sorghum bicolor)、菠菜属物种(Spmacia spp.)、Sporobolus fimbriatus、Stiburus alopecuroides、Stylosanthos humilis、葫芦茶属物种(Tadehagispp)、落羽杉(Taxodium distichum)、阿拉伯黄背草(Themeda triandra)、车轴草属物种(Trifolium spp.)、小麦属物种(Triticum spp.)、异叶铁杉(Tsuga heterophylla)、越桔属物种(Vaccinium spp.)、野豌豆属物种(Viciaspp.)、葡萄(Vitis vinifera)、锥穗沃森花(Watsonia pyramidata)、马蹄莲(Zantedeschia aethiopica)、玉米(Zea mays)、苋属(amaranth)、菊芋(artichoke)、天门冬属(asparagus)、青花椰菜(broccoli)、孢子甘蓝(Brusselssprouts)、甘蓝、卡诺拉油菜(canola)、胡萝卜、花椰菜、芹菜、羽衣甘蓝(collard greens)、亚麻、球茎甘蓝(kale)、兵豆属(lentil)、油菜(oilseed rape)、秋葵(okra)、洋葱、马铃薯、稻、大豆、草莓、甜菜、甘蔗、向日葵、西红柿、南瓜(squash)、茶和藻类等。
优选的,植物为作物植物,诸如大豆、向日葵、卡诺拉油菜、苜蓿、油菜、棉花、番茄、马铃薯或烟草的作物植物。进一步优选地,植物为单子叶植物,例如甘蔗。更加优选地,植物是谷类,例如稻、玉米、小麦、大麦、小米、黑麦、高粱或燕麦。
I型DnaJ样多肽的活性可以通过提高多肽的水平来增加。可选的,当I型DnaJ样多肽的水平没有改变,或者甚至是I型DnaJ样多肽的水平降低时,也可以增加活性。这可以发生在当多肽的内在性质改变时,例如通过制备比野生型多肽更活跃的突变体形式。本文提及的“优先”增加活性意指在非胁迫条件下生长的植物细胞溶胶中I型DnaJ样多肽或其同源物的活性的靶向增加,该活性增加超过了在相当条件下生长的野生型植物的细胞溶胶中发现的增加。
本文所定义的术语“I型DnaJ样多肽或其同源物”是指在其羧基末端包含CaaX基序的I型DnaJ样多肽。
所有DnaJ蛋白质的共同点在于J结构域的存在,其由大约70个氨基酸组成(通常位于蛋白质的氨基末端)并且在其中间包含高度保守的HPD三肽(InterPro参考IPR001623;Zdobnov等人,(2002)18(8):1149-50);由4个α螺旋组成的“J”结构域与Hsp70蛋白质相互作用。在拟南芥基因组中,已经鉴定了至少89种包含J结构域的蛋白质(Miernyk(2001)CellStress&Chaperones)。目前已经鉴定了18种Hsp70蛋白质。
I型DnaJ结构域蛋白质(或DnaJ蛋白质)通过下列(从氨基末端到羧基末端)结构域的存在来进一步表征:
1)大约30个氨基酸残基的G/F结构域区域,其富含甘氨酸(G)和苯丙氨酸(F),其用于调节靶标多肽特异性;以及
2)包含4个重复CXXCXGXG的富含Cys的锌指结构域,其中X代表了带电或极性残基;这4个重复片段成对作用来形成锌结合结构域I和II(InterPro参考IPR001305;Linke等人(2003)J Biol Chem 278(45):44457-44466);认为锌指结构域介导直接的蛋白质:蛋白质相互作用并且更特异性的结合待递送到Hsp70的非天然多肽;
3)羧基末端结构域(CTD;InterPro参考IPR002939)。
在其天然形式(可以是可溶的或膜结合的形式)中,DnaJ蛋白质可以靶向到多个亚细胞区室。用在本发明方法中的DnaJ蛋白质是没有信号序列或转运肽的那些蛋白质,并且因此主要位于植物细胞的细胞溶胶中。
用在本发明方法中的DnaJ蛋白质是在其羧基末端包含CaaX基序的那些蛋白质。此多肽因此以可溶的或膜结合的形式存在,每种形式的存在可以互换。
使用本领域熟知的常规技术可以容易的鉴定“I型DnaJ样多肽或其同源物”。例如如在Zhou等人,(2000),Protein Expression&Purification 19:253-258中描述的,可以容易的体外确定DnaJ对DnaK的ATP酶活性的刺激。DnaJ促进DnaK和非天然多肽(如变性的荧光素酶)之间的复合物形成的能力可以通过ELISA确定(Fan等人,(2004)Molec.Biol.Cell 15:761-773)。
使用序列比对可以容易的鉴定出“I型DnaJ样多肽或其同源物”。用于比较的序列比对方法是本领域熟知的,此类方法包括GAP、BESTFIT、BLAST、FASTA和TFASTA。GAP使用Needleman和Wunsch((1970)J.Mol.Biol.48:443-453)的算法来寻找两个全序列的比对(使匹配数最大并且空位数最小)。BLAST算法(Altschul等人,(1990)J Mol Biol 215:403-10)计算序列同一性百分比并且进行两个序列之间相似性的统计学分析。进行BLAST分析的软件是通过National Centre for Biotechnology Information公开可用的。I型DnaJ样多肽的同源物和它们对SEQ ID NO:2所代表的用在本发明方法中的I型DnaJ样氨基酸序列的同一性百分比可以使用例如基于改良的ClustalW算法(InforMax,Bethesda,MD,http://www.informaxinc.com)的VNTI AlignX多重对比程序(具有空位开放罚分以及空位延伸的缺省设置)来容易的鉴定。优选的,在其羧基末端包含CaaX基序,并且在本发明方法中是有用的I型DnaJ样多肽或其同源物与SEQ ID NO:2具有一定的同一性,所述序列同一性按照增加的优选顺序为至少35%、40%、45%、50%、55%、60%、65%、70%、75%、80%、85%、90%或95%。
术语“I型DnaJ样多肽或其同源物”所包括的多肽实例列在表1中,为SEQ ID NO:2、SEQ ID NO:4、SEQ ID NO:6、SEQ ID NO:8、SEQ IDNO:10、SEQ ID NO:12、SEQID NO:14、SEQ ID NO:16、SEQ ID NO:18、SEQ ID NO:20、SEQ ID NO:22、SEQ ID NO:24、SEQ ID NO:26、SEQ ID NO:28、SEQ ID NO:30、SEQ ID NO:32、SEQ ID NO:34、SEQID NO:36、SEQ ID NO:38、SEQ ID NO:40、SEQ ID NO:42、SEQ ID NO:44、SEQ ID NO:46、SEQ ID NO:48、SEQ ID NO:50、SEQ ID NO:52、SEQ ID NO:54和SEQ ID NO:56。用来例示性说明本发明的I型DnaJ样多肽序列由SEQ ID NO:2所代表。
表1:来自不同生物的编码I型DnaJ样多肽(蛋白质SEQ ID NO)的核酸(DNA SEQ ID)实例
基因名 | 登录号DNA | DNA SEQ ID | 登录号蛋白质 | 蛋白质SEQID | 来源 |
CDS1877DnaJ | AK066420.1 | SEQ ID NO:1 | VT | SEQ ID NO:2 | 稻(Oryza sativa) |
Orysa_DNAJII CAAX | AK10195 | SEQ ID NO:3 | VT | SEQ ID NO:4 | 稻 |
Orysa_DNAJHI CAAX | AK105028 | SEQ ID NO:5 | VT | SEQ ID NO:6 | 稻 |
Orysa_DNAJIV CAAX | AK104315 | SEQ ID NO:7 | VT | SEQ ID NO:8 | 稻 |
Zeama_ZMDJ1 | BT016805* | SEQ ID NO:9 | T01643 | SEQ ID NO:10 | 玉米 |
Zeama_DNAJ I CAAX | AY103727.1 | SEQ ID NO:11 | VT | SEQ ID NO:12 | 玉米 |
Zeama_DNAJ II CAAX | AY108160.1 | SEQ ID NO:13 | VT | SEQ ID NO:14 | 玉米 |
Triae_DNAJ | BT008914.1 | SEQ ID NO:15 | VT | SEQ ID NO:16 | 玉米 |
At5g22060AtJ2 CAAX | L36113 | SEQ ID NO:17 | AAB8679 | SEQ ID NO:18 | 拟南芥 |
At3g44110AtJ3 CAAX | NM_114279 | SEQ ID NO:19 | S71199 | SEQ ID NO:20 | 拟南芥 |
Atrnu_DNAJ | L09124 | SEQ ID NO:21 | VT | SEQ ID NO:22 | 大洋洲滨藜(Atriplexnummularia) |
Cucsa_DNAJ-1 | X67695 | SEQ ID NO:23 | VT | SEQ ID NO:24 | 黄瓜(Cucumissativus) |
Dauca_J1P | AF308737 | SEQ ID NO:25 | VT | SEQ ID NO:26 | 胡萝卜(Daucuscarota) |
Glyma_pm37DNAJ | AF169022 | SEQ ID NO:27 | VT | SEQ ID NO:28 | 大豆(Glycinemax) |
Hevbr_DNAJ | AF085275 | SEQ ID NO:29 | AAD1205 | SEQ ID NO:30 | 橡胶(Heveabrasiliensis) |
Lyces_DNAJ | AF124139 | SEQ ID NO:31 | AAF28382 | SEQ ID NO:32 | 番茄(Lycopersicumesculentum) |
Medsa_DNAJ | AJ000995 | SEQ ID NO:33 | CAA0444 | SEQ ID NO:34 | 紫花苜蓿(Medicagosativa) |
Nicta_DNAJ | AJ299254 | SEQ ID NO:35 | CAC1282 | SEQ ID NO:36 | 烟草(Nicotianatabacum) |
Salgi_DNAJ2 | AB003137 | SEQ ID NO:37 | BAA7688 | SEQ ID NO:38 | 河柳(Salixgilgiana |
Salgi_DNAJ | AB015601 | SEQ ID NO:39 | BAA3512 | SEQ ID NO:40 | 河柳 |
Solto_DNAJ | X94301 | SEQ ID NO:41 | CAA6396 | SEQ ID NO:42 | 马铃薯(Solanumtuberosum) |
Orysa_DNAJCASQ | AK110691 | SEQ ID NO:43 | VT | SEQ ID NO:44 | 稻 |
Triae_DNAJII CASQ | BT009366 | SEQ ID NO:45 | VT | SEQ ID NO:46 | 小麦(Triticumaestivum) |
Ceael_DNaJ | NM_072051 | SEQ ID NO:47 | NP_50445 | SEQ ID NO:48 | Caenorhabditiselegans |
Homsa_HsJ2 | D13388 | SEQ ID NO:49 | P31689 | SEQ ID NO:50 | 人(Homosapiens) |
Sacce_YDJ1 | NC_001146 | SEQ ID NO:51 | NP_01433 | SEQ ID NO:52 | 酿酒酵母(Saccharomycescereviseae) |
Homsa_DNAJA2 | NM_005880 | SEQ ID NO:53 | NP_00587 | SEQ ID NO:54 | 人 |
Musmu_mDj3 | NM_019794 | SEQ ID NO:55 | Q9QYJ0 | SEQ ID NO:56 | 小鼠(Musmusculus) |
VT=有效翻译;*具有小修正
要理解落入“I型DnaJ样多肽或其同源物”定义的序列不限于列在表1中的SEQ ID NO:2、SEQ ID NO:4、SEQ ID NO:6、SEQ ID NO:8、SEQ ID NO:10、SEQ ID NO:12、SEQ ID NO:14、SEQ ID NO:16、SEQID NO:18、SEQ ID NO:20、SEQ ID NO:22、SEQ ID NO:24、SEQ ID NO:26、SEQ ID NO:28、SEQ ID NO:30、SEQ ID NO:32、SEQ ID NO:34、SEQ ID NO:36、SEQ ID NO:38、SEQ ID NO:40、SEQ ID NO:42、SEQID NO:44、SEQ ID NO:46、SEQ ID NO:48、SEQ ID NO:50、SEQ ID NO:52、SEQ ID NO:54和SEQ ID NO:56,而是在其羧基末端包含CaaX基序的任何I型DnaJ样多肽均适合用于本发明方法中。
编码I型DnaJ样多肽或其同源物的核酸可以是任何天然或合成核酸。因此,如本文定义的术语“I型DnaJ样核酸/基因”是编码上述定义的I型DnaJ样多肽或其同源物的任何核酸/基因。I型DnaJ样核酸的实例包括列在表1中的那些,如SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:5、SEQ ID NO:7、SEQ ID NO:9、SEQ ID NO:11、SEQ ID NO:13、SEQ IDNO:15、SEQ ID NO:17、SEQ ID NO:19、SEQ ID NO:21、SEQ ID NO:23、SEQ ID NO:25、SEQ ID NO:27、SEQ ID NO:29、SEQ ID NO:31、SEQ ID NO:33、SEQ ID NO:35、SEQ ID NO:37、SEQ ID NO:39、SEQID NO:41、SEQ ID NO:43、SEQ ID NO:45、SEQ ID NO:47、SEQ ID NO:49、SEQ ID NO:51、SEQ ID NO:53和SEQ ID NO:55。I型DnaJ样核酸/基因及其变体适合用于本发明方法中。I型DnaJ样核酸/基因的变体包括I型DnaJ样核酸/基因的部分,和/或能够与I型DnaJ样核酸/基因杂交的核酸。
如本文定义的术语部分是指包含至少600个核苷酸的DNA片段,该部分编码至少200个氨基酸的多肽,所述多肽从氨基末端到羧基末端包含DnaJ结构域、富含G/F结构域和富含Cys锌指结构域、CTD结构域和CaaX基序。优选的,该部分包含至少1050个核苷酸,编码至少350个氨基酸的多肽,所述多肽从氨基末端到羧基末端包含DnaJ结构域、富含G/F结构域、富含Cys锌指结构域、CTD结构域和CaaX基序。更优选的,上述定义的部分是表1中SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:5、SEQ ID NO:7、SEQ ID NO:9、SEQ ID NO:11、SEQ ID NO:13、SEQ IDNO:15、SEQ ID NO:17、SEQ ID NO:19、SEQ ID NO:21、SEQ ID NO:23、SEQ ID NO:25、SEQ ID NO:27、SEQ ID NO:29、SEQ ID NO:31、SEQ ID NO:33、SEQ ID NO:35、SEQ ID NO:37、SEQ ID NO:39、SEQID NO:41、SEQ ID NO:43、SEQ ID NO:45、SEQ ID NO:47、SEQ ID NO:49、SEQ ID NO:51、SEQ ID NO:53或SEQ ID NO:55任一个所代表的核酸的部分。
例如,通过对I型DnaJ样核酸产生一个或多个缺失来制备部分。除去多核苷酸序列的一个实例包括编码特异性亚细胞导向序列的序列,如线粒体或质体导向序列。部分可以以分离的形式使用或它们可以融合到其他编码(或非编码)序列中,从而(例如)产生组合了一些活性的蛋白质。当融合到其他编码序列时,翻译产生的所得多肽可以大于为I型DnaJ样片段所预测的。例如,可以将编码法尼基化基序的寡核苷酸融合到编码最初缺乏此基序的I型DnaJ样多肽的I型DnaJ样多核苷酸序列。可以将部分融合到I型DnaJ家族其他成员的编码序列的其他部分,从而在2个最初I型DnaJ样多肽之间互换结构域。例如,一种I型DnaJ样多肽的CTD结构域可以与另一种I型DnaJ样多肽的CTD结构域交换。
I型DnaJ样核酸/基因的另一变体是能够在降低的严格条件下(优选在严格条件下)与本文上述定义的I型DnaJ样核酸/基因杂交的核酸,其中杂交序列至少编码I型DnaJ样多肽的J结构域和富含Cys锌指结构域。优选的,此类变体包含表现了I型DnaJ样多肽特征的所有结构域,从氨基末端到羧基末端的DnaJ结构域、富含D/F结构域、富含Cys锌指结构域和CTD结构域,以及额外的CaaX基序。
优选的,杂交序列是能够与下列代表的核酸,或者如本文上述定义的任何前述序列的部分杂交的序列:表1的SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:5、SEQ ID NO:7、SEQ ID NO:9、SEQ ID NO:11、SEQ IDNO:13、SEQ ID NO:15、SEQ ID NO:17、SEQ ID NO:19、SEQ ID NO:21、SEQ ID NO:23、SEQ ID NO:25、SEQ ID NO:27、SEQ ID NO:29、SEQ ID NO:31、SEQ ID NO:33、SEQ ID NO:35、SEQ ID NO:37、SEQID NO:39、SEQ ID NO:41、SEQ ID NO:43、SEQ ID NO:45、SEQ ID NO:47、SEQ ID NO:49、SEQ ID NO:51、SEQ ID NO:53或SEQ ID NO:55种任一个。
本文所定义的术语“杂交”是基本同源的互补核苷酸序列相互退火的过程。杂交过程可以完全发生在溶液中,即互补核酸都在溶液中。杂交过程还可以如此进行,即其中互补核酸之一固定于基质如磁珠、琼脂糖珠子或任何其他树脂上。此外杂交过程还可以如此进行,即其中互补核酸之一固定于固相支持物如硝化纤维素膜或尼龙膜上,或通过例如照相平版印刷术固定到例如硅酸玻璃支持物上(后者称之为核酸阵列或微阵列,或称之为核酸芯片)。为了使得杂交发生,通常使核酸分子热变性或化学变性,以使双链解链成两条单链和/或从单链核酸中除去发夹或其他二级结构。杂交的严格性受诸如温度、盐浓度和杂交缓冲液组成等条件的影响。
在诸如Southern杂交和Northern杂交的核酸杂交实验中,“严格杂交条件”和“严格杂交洗涤条件”是序列依赖性的,并且在不同的环境参数下不同。本领域技术人员认识到在杂交和洗涤中可以改变多种参数,并且所述改变将保持或改变严格条件。
Tm是在确定的离子强度和pH下,50%的目的序列与完全匹配的探针杂交时的温度。Tm依赖于溶液条件、碱基组成以及探针的长度。例如,较长的序列在较高温度下特异杂交。杂交的最大速率是在Tm低于大约16℃到32℃下得到的。杂交溶液中一价阳离子的存在降低了两条核酸链之间的静电排斥,从而促进杂交体的形成;对于高达0.4M的钠浓度,这个作用是明显的。甲酰胺降低了DNA-DNA和DNA-RNA双链体的解链温度,每百分数甲酰胺降低0.6至0.7℃,加入50%的甲酰胺虽然使杂交速率降低,但使杂交在30至45℃进行。碱基对错配降低了杂交速率和双链体的热稳定性。一般对于大探针,每%碱基错配使Tm降低大约1℃。取决于杂交体的类型,Tm可以使用下列公式计算:
1.DNA-DNA杂交体(Meinkoth和Wahl,Anal.Biochem.,138:267-284,1984):
Tm=81.5℃+16.6×log[Na+]a+0.41×%[G/Cb]-500×[Lc]-1-0.61×%甲酰胺
2.DNA-RNA或RNA-RNA杂交体:
Tm=79.8+18.5(log10[Na+]a)+0.58(%G/Cb)+11.8(%G/Cb)2-820/Lc
3.寡-DNA或寡-RNAd杂交体:
对于<20个核苷酸:Tm=2(ln)
对于20-35个核苷酸:Tm=22+1.46(ln)
a或对于其他一价阳离子,但是仅仅在0.01-0.4M的范围精确。
b仅仅对30%至75%范围的%GC精确。
cL=双链体中碱基对的长度。
d寡,寡核苷酸;ln,引物的有效长度=2×(G/C数目)+(A/T数目)。
注解:对于每1%甲酰胺,Tm降低大约0.6至0.7℃,而6M尿素的存在降低Tm大约30℃。
杂交的特异性通常是杂交后洗涤的函数。为了除去非特异性杂交产生的背景,用稀释的盐溶液洗涤样品。此类洗涤的关键因素包括最终洗涤溶液的离子强度和温度:盐浓度越低以及洗涤温度越高,洗涤的严格性越高。洗涤条件一般在杂交严格性或低于杂交严格性下进行。一般而言,用于核酸杂交试验或基因扩增检测方法的合适的严格条件如上所述。也可以选择更高或更低的严格条件。一般而言,低严格条件选择为比在确定的离子强度和pH下特异性序列的热解链温度(Tm)低大约50℃。中等严格条件是当温度比Tm低20℃时,而高严格条件是当温度比Tm低10℃时。例如,严格条件是那些至少,例如条件A-L的严格性;并且简化的严格条件是至少,例如条件M-R的严格性。非特异性的结合可以使用一些已知技术中的任一种(例如用含有蛋白质的溶液将膜封闭,添加异源RNA、DNA和SDS到杂交缓冲液中,以及周RNA酶处理)来调控。杂交和洗涤条件的实例列在下列表2中。
表2:杂交和洗涤条件的实例
严格条件 | 多核苷酸杂交体± | 杂交体长度(bp) | 杂交温度和缓冲液 | 洗涤温度和缓冲液 |
A | DNA:DNA | >或等于50 | 65℃ 1×SSC;or 42℃,1×SSC和50%甲酰胺 | 65℃;0.3×SSC |
B | DNA:DNA | <50 | Tb*;1×SSC | Tb*;1×SSC |
C | DNA:RNA | >或等于50 | 67℃ 1×SSC;or 45℃,1×SSC和50%甲酰胺 | 67℃;0.3×SSC |
D | DNA:RNA | <50 | Td*;1×SSC | Td*;1×SSC |
E | RNA:RNA | >或等于50 | 70℃ 1×SSC;or 50℃,1×SSC和50%甲酰胺 | 70℃;0.3×SSC |
F | RNA:RNA | <50 | Tf*;1×SSC | Tf*;1×SSC |
G | DNA:DNA | >或等于50 | 65℃ 4×SSC;or 45℃,4×SSC和50%甲酰胺 | 65℃;1×SSC |
H | DNA:DNA | <50 | Th*;4×SSC | Th*;4×SSC |
I | DNA:RNA | >或等于50 | 67℃ 4×SSC;or 45℃,4×SSC和50%甲酰胺 | 67℃;1×SSC |
J | DNA:RNA | <50 | Tj*;4×SSC | Tj*;4×SSC |
K | RNA:RNA | >或等于50 | 70℃ 4×SSC;or 40℃,6×SSC和50%甲酰胺 | 67℃;1×SSC |
L | RNA:RNA | <50 | T1*;2×SSC | T1*;2×SSC |
M | DNA:DNA | >或等于50 | 50℃ 4×SSC;or 40℃,6×SSC和50%甲酰胺 | 50℃;2×SSC |
N | DNA:DNA | <50 | Tn*;6×SSC | Tn*;6×SSC |
O | DNA:RNA | >或等于50 | 55℃ 4×SSC;or 42℃,6×SSC和50%甲酰胺 | 55℃;2×SSC |
P | DNA:RNA | <50 | Tp*;6×SSC | Tp*;6×SSC |
Q | RNA:RNA | >或等于50 | 60℃ 4×SSC;or 45℃,6×SSC和50%甲酰胺 | 60℃.;2×SSC |
R | RNA:RNA | <50 | Tr*;4×SSC | Tr*;4×SSC |
“杂交体长度”是杂交核酸的预期长度。当已知序列的核酸杂交时,可以通过比对序列和鉴别本文所述的保守区来确定杂交体长度。
在杂交和洗脱缓冲液中可以用SSPE(1×SSPE是0.15 M NaCl、10mM NaH2PO4和1.25 mM EDTA,pH 7.4)代替SSC(1×SSC是0.15 MNaCl和15 mM柠檬酸钠);在杂交完成后洗脱15分钟。杂交和洗脱可以另外地包括5×Denhardt试剂、0.5-1.0%SDS、100μg/ml变性的片段化的鲑精DNA、0.5%焦磷酸钠和高达50%甲酰胺。
*Tb-Tr:对于预期长度少于50个碱基对的杂交体,杂交温度应该比杂交体的熔解温度Tm低5-10℃,根据上述公式确定Tm。
±本发明还包括用PNA或修饰的核酸取代任意一个或多个DNA或RNA杂交体配偶体。
为了定义严格性水平的目的,通常参考Sambrook等人(2001)Molecular Cloning:a laboratory manual,第3版,Cold Spring HarborLaboratory Press,CSH,New York或Current Protocols in MolecularBiology,John Wiley&Sons,N.Y.(1989)。
I型DnaJ样核酸或其变体可以来自任何天然或人工来源。可以通过考虑周到的人为操作,从在组合物和/或基因组环境中的此核酸的天然形式修饰它。核酸是原核或真核来源的,来自微生物来源如酵母或真菌,或来自植物、藻类或动物(包括人)来源。优选的,核酸是真核来源的。核酸另外优选是植物来源的,既可以来自相同的植物物种(例如与其要被引入的物种相同)又可来自不同的植物物种。核酸可以分离自单子叶植物物种,优选来自禾本科(Poaceae),更优选来自稻。更优选的,分离自稻的I型DnaJ样核酸是由SEQ ID NO:1所代表的并且I型DnaJ样氨基酸序列是由SEQIDNO:2所代表的。
I型DnaJ样多肽或其同源物的活性可以通过引入遗传修饰(优选在I型DnaJ样基因的基因座中)来增加。本文中所定义的基因的基因座意指包括目的基因和编码区上游或下游10kb的基因组区域。
例如,遗传修饰可以通过以下的任何一种(或多种)方法引入:TDNA激活、TILLING、定点诱变、定向进化和同源重组,或通过在植物细胞细胞溶胶中引入和/或表达编码I型DnaJ样多肽或其同源物的核酸,该I型DnaJ样多肽在其羧基末端包含CaaX基序。在引入遗传修饰后,接下来的步骤是选择在植物细胞溶胶中I型DnaJ样多肽增加的活性,这种活性的增加使得植物在非胁迫条件下具有增加的植物产量。
T-DNA激活标签(Hayashi等人,Science(1992),1350-1353)涉及在目的基因的基因组区域或基因编码区域上游或下游10KB中插入通常包含启动子(也可以是翻译增强子或内含子)的T-DNA,如此构型以便启动子指引靶基因表达。通常,破坏靶基因天然启动子对其的表达调控,而使该基因处于新引入的启动子的控制之下。该启动子一般嵌入到T-DNA中。例如,该T-DNA通过农杆菌属感染随机插入到植物基因组中,并导致靠近该插入T-DNA的基因过量表达。由于接近引入的启动子的基因过量表达,所得的转基因植物显示出显性表型。待引入的启动子可以是能指导基因在所需的生物体(在本案中为植物)中表达的任何启动子。例如,组成型、组织偏好的、细胞类型特异性的以及诱导型启动子都适合用于T-DNA激活。优选的,启动子是能够在植物种子组织中推动基因表达的启动子。
也可以使用TILLING技术(靶向诱导基因组局部突变技术),把遗传修饰引入到I型DnaJ样基因的基因座中。这是一种诱变技术,可用于产生和/或鉴定,并最终分离能显示DnaJ样活性的I型DnaJ样核酸诱变变体。TILLING还能够选择携带这类突变变体的植物。这些突变变体甚至可表现出比该基因的天然形式所表现出的更高的DnaJ样活性。TILLNG将高密度诱变与高通量筛选方法结合起来。TILLING一般遵循的步骤是:(a)EMS诱变(Redei GP和Koncz C(1992)In Methods in Arabidopsis Research,Koncz C,Chua NH,Schell J编著,新加坡,World Scientific Publishing Co,第16-82页;Feldmann等人,(1994)In Meyerowitz EM,Somerville CR编著,Arabidopsis.Cold Spring Harbor Laboratory Press,Cold SpringHarbor,NY,第137-172页;Lightner J和Caspar T(1998)In JMartinez-Zapater,J Salinas编著,Methods on Molecular Biology,第82卷.Humana Press,Totowa,NJ,第91-104页);(b)DNA制备和个体合并;(c)目标区域的PCR扩增;(d)变性和退火以允许形成异源双链体;(e)DHPLC,其中库中异源双链体的存在检测为色谱图中额外的峰值;(f)鉴定突变个体;和(g)突变PCR产物的测序。TILLING的方法是本领域熟知的(McCallum等人,(2000)Nat Biotechnol 18:455-457;Stemple(2004)NatRev Genet 5(2):145-50综述)。
定点诱变可用于产生I型DnaJ样核酸或其部分的变体。一些方法可用来实现定点诱变,最常用方法是基于PCR的方法(Current Protocols inMolecular Biology,Wiley编著http://www.4ulr.com/products/currentprotocols/index.html)。
由基因DNA改组的重复、随后是适当的筛选和/或选择组成的定向进化也可以用来生成下列的变体:编码具有增加的生物学活性的I型DnaJ样多肽、或其同源物、或其部分的I型DnaJ样核酸或其部分(Castle等人,(2004)Science 304(5674):1151-4;美国专利5,811,238和6,395,547)。
T-DNA激活、TILLING、定点诱变和定向进化是能够产生新的等位基因和I型DnaJ样变体的技术实例。
同源重组能够在基因组的指定选择位置引入选定的核酸。同源重组是生物学中通常使用的标准技术,用于较低等的生物如酵母或苔藓physcomitrella。用于在植物中进行同源重组的方法不仅已在模式植物中描述(Offringa等人,1990 EMBO J.9(10):3077-84),而且在作物植物例如稻中描述(Terada等人,(2002)Nat Biotech 20(10):1030-4;Iida和Terada(2004)Curr Opin Biotech 15(2):132-8)。例如,待靶向的核酸(其可以是如上文所定义的I型DnaJ样核酸或其变体)靶向到I型DnaJ样基因的基因座。待靶向的核酸可以是改良的等位基因,用于替换内源基因,或另外引入到内源基因中。
根据本发明优选的实施方案,可以通过在植物细胞溶胶中引入和/或表达编码I型DnaJ样多肽或其同源物的核酸来增加植物产量,该I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序。
引入遗传修饰(在本案中不需要在I型DnaJ样基因的基因座中)的优选的方法是在植物细胞溶胶中引入和/或表达编码I型DnaJ样多肽或其同源物的外源核酸,该外源I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序。待引入到植物中的核酸可以是如前文所定义的全长核酸,或可以是如前文所定义的部分或杂交序列。
本文定义的术语“外源”是指分离的基因/核酸,其可以是来自相同或不同的植物物种,例如根据上述定义,在稻植物中引入和/或表达的分离的稻基因/核酸是“外源的”。
蛋白质的“同源物”包含相对所讨论的未修饰蛋白质具有氨基酸取代、缺失和/或插入,并且具有与它们所衍生自的未修饰蛋白质类似的生物学和功能活性的肽、寡肽、多肽、蛋白质和酶。要产生此类同源物,蛋白质的氨基酸可以用其他具有相似性质(例如相似的疏水性、亲水性、抗原性、形成或断裂α-螺旋结构或β-片层结构的倾向性)的氨基酸代替。保守取代表为本领域众所周知(例如见Creighton(1984)Proteins,W.H.Freeman andCompany)。下表3给出了保守氨基酸取代的实例。
表3:保守性氨基酸取代的实例:
残基 | 保守性取代 | 残基 | 保守性取代 |
Ala | Ser | Leu | Ile;Val |
Arg | Lys | Lys | Arg;Gln |
Asn | Gln;His | Met | Leu;Ile |
Asp | Glu | Phe | Met;Leu;Tyr |
Gln | Asn | Ser | Thr;Gly |
Cys | Ser | Thr | Ser;Val |
Glu | Asp | Trp | Tyr |
Gly | Pro | Tyr | Trp;Phe |
His | Asn;Gln | Val | Ile;Leu |
Ile | Leu,Val |
同源物包括直系同源物和旁系同源物,它们涉及用于描述基因遗传关系的进化概念。旁系同源物是在相同物种中通过祖先基因复制产生的基因,而直系同源物是不同生物体中因物种形成所致的基因。
例如,单子叶植物物种中的直系同源物可以通过实施所谓的交互blast搜索很容易地找到。这可以通过第一次blast完成,其包括在任何序列数据库,如可在http://www.ncbi.nlm.nih.gov中找到的公众可用的NCBI数据库中,对所讨论的序列(例如,SEQ ID NO:1或SEQ ID NO:2)进行blast。当起始于核苷酸序列时可以使用BLASTN或TBLASTX(使用标准缺省值),并且当起始于蛋白质序列时可以使用BLASTP或TBLASTN(使用标准缺省值)。可选的过滤blast结果。之后将过滤结果或未过滤结果的全长序列对所讨论的序列来源的生物序列进行反向BLAST(第二次BLAST)(其中所讨论序列是SEQ ID NO:1或SEQ ID NO:2,则第二次blast应当针对稻序列)。然后对第一次和第二次BLAST的结果进行比较。如果第二次blast的高级别目标(hit)是来自与所讨论序列来源物种相同的物种,则鉴定了旁系同源物;如果高级别目标不是来自与所讨论序列来源物种相同的物种,则鉴定了直系同源物。高级别目标是具有低E值的那些。E值越低,分数越显著(或者换句话说,偶然发现目标的机会越低)。E值的计算是本领域熟知的。在大家族的情况下,使用ClustalW,接着使用相邻连接树以帮助观察相关基因的聚类并且鉴定直系同源物和旁系同源物。
同源物可以是蛋白质“取代变体”的形式,即其中氨基酸序列中的至少一个残基已经被除去,并且不同的残基插入到它的位置上。氨基酸取代一般是单残基的,但是取决于对多肽构成的功能限制可以是成串的;插入片段通常是大约1到10个氨基酸残基。优选的,氨基酸取代包括保守性氨基酸取代。
同源物还可以是蛋白质的“插入变体”的形式,即将其中一个或多个氨基酸残基引入到蛋白质中的预定位点。插入可包括氨基末端和/或羧基末端的融合,以及单个或多个氨基酸的序列内插入。通常,氨基酸序列内的插入将小于氨基末端或羧基末端的融合,约为1到10个残基的数量。氨基末端或羧基末端融合蛋白质或肽的实例包括如用于酵母双杂交系统的转录激活因子的结合结构域或激活结构域、噬菌体外壳蛋白、(组氨酸)6-标签、谷胱甘肽S-转移酶-标签、A蛋白、麦芽糖结合蛋白、二氢叶酸还原酶、Tag·100表位、c-myc表位、FLAG表位、lacZ、CMP(钙调蛋白结合肽)、HA表位、C蛋白表位和VSV表位。
蛋白质“缺失变体”形式的同源物的特征在于从该蛋白质除去一个或多个氨基酸。此类缺失变体的一个实例是从I型DnaJ样蛋白质中移除线粒体或质体的导向序列,否则其会靶向到这些器官。
使用本领域熟知的肽合成技术,如固相肽合成等等,或通过重组DNA操作,可以很容易的制备蛋白质的氨基酸变体。对DNA序列进行操作以产生蛋白质的取代、插入或缺失变体的方法是本领域熟知的。例如,在DNA的预定位点产生取代突变的技术是本领域技术人员熟知的,包括M13诱变、T7-Gen体外诱变(USB,Cleveland,OH)、快速变换的定点诱变(Stratagene,San Diego,CA)、PCR-介导的定点诱变或其他定点诱变方案。
I型DnaJ样多肽或其同源物可以是衍生物。“衍生物”包括肽、寡肽、多肽、蛋白质和酶,与蛋白质天然存在形式的氨基酸序列相比,例如,如SEQ ID NO:2所示的,其可以包括天然和非天然存在氨基酸残基的取代、缺失或添加。蛋白质的“衍生物”包括肽、寡肽、多肽、蛋白质和酶,与多肽天然存在形式的氨基酸序列相比,其可以包括天然存在的改变的、糖基化的、酰基化的、异戊二烯化的氨基酸残基或非天然存在的氨基酸残基。与其所衍生自的氨基酸序列相比,衍生物还可包括一个或多个非氨基酸取代基,例如与氨基酸序列共价或非共价结合的报道分子或其他配体,如结合以便于其检测的报道分子,以及相对于天然存在蛋白质的氨基酸序列而言非天然存在的氨基酸残基。
I型DnaJ样多肽或其同源物可以是I型DnaJ样核酸/基因的可变的剪接变体所编码的。本文所用的术语“可选择的剪接变体”涵盖这样的核酸序列变体,即其中选定的内含子和/或外显子已经切除、置换或添加,或者其中内含子已经缩短或延长。这类变体是其中蛋白质的生物学活性保持的那些变体,其可以通过选择性的保留蛋白质的功能片段来获得。此类剪接变体可以是天然发现的或可以是人造的。用于产生这种剪接变体的方法是本领域熟知的。优选的是编码作为I型DnaJ样多肽或其同源物的多肽的剪接变体,该I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序,特别是SEQ ID NO:1的剪接变体。
同源物还可以是由编码I型DnaJ样多肽或其同源物的核酸的等位变体所编码的,所述等位变体优选是SEQ ID NO:1所代表的核酸的等位变体。更优选的,等位变体编码的多肽是I型DnaJ样多肽或其同源物,该I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序。
等位变体天然存在,并且包括在本发明方法中的是这些天然等位基因的用途。等位变体包括单核苷酸多态性(SNP)以及小的插入/缺失多态性(INDEL)。INDEL的大小通常小于100bp。在大多数生物天然存在的多态性品系中SNP和INDEL形成了最大的一类序列变体。
根据本发明优选的方面,设想了I型DnaJ样核酸或其变体的增强或增加的表达。得到基因或基因产物增强或增加的表达的方法详细记录在本领域中并且包括例如适当启动子驱动的过量表达,转录增强子或翻译增强子的使用。可以将作用为启动子或增强子元件的分离的核酸引入至多核苷酸非异源形式的适当位置上(一般为上游),从而上调I型DnaJ样核酸或其变体的表达。例如,可以通过突变、缺失和/或取代来体内改变内源启动子(参见Kmiec,美国专利5,565,350号;Zarling等人,PCT/US93/03868),或者可以将分离的启动子以适当的方向和与本发明基因的距离引入植物细胞中,从而调控基因的表达。
如果希望多肽表达,通常需要在多核苷酸编码区的3’-端包含聚腺苷酸化区域。聚腺苷酸化区域可以来自天然基因、来自多种其他植物基因或来自T-DNA。待添加的3’端序列可以来自例如胭脂碱合酶或章鱼碱合酶基因,或可选的来自其他植物基因,或较小优选的来自任何其他真核基因。
还可以将内含子序列添加到部分编码序列的5’非翻译区或编码序列以增加在细胞溶胶中聚集的成熟信息的量。已经表明在植物和动物表达构建体的转录单位中包含可剪接的内含子,能在mRNA和蛋白质水平上使基因表达增加达1000倍,Buchman和Berg,Mol.Cell Biol.8,4395-4405(1988);Callis等人,Genes Dev.1,1183-1200(1987))。当将内含子置于转录单位的5’端附近时,基因表达的此类内含子增强通常是最大的。本领域已知玉米内含子Adh1-S内含子1、2和6,Bronze-1内含子的用途。总体上参见TheMaize Handbook,116章,Freeling和Walbot编著,Springer,N.Y.(1994)。
本发明还提供了遗传构建体和载体以便于引入和/或表达用在本发明方法中的核苷酸序列。
因此,提供了基因构建体,其包含:
(i)编码I型DnaJ样多肽或其同源物的核酸或其变体,该I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序;以及
(ii)一种或多种能够驱动(i)的核酸序列表达的调控序列;以及任选的
(iii)转录终止序列。
本发明还提供了如本文所定义的构建体的用途,其用于在非胁迫条件下生长的植物中增加植物产量的方法中。
可用于本发明方法的构建体可以使用本领域技术人员熟知的重组DNA技术构建。基因构建体可插入到载体中,该载体可以是商购的,适于转化到植物中并适于在转化的细胞中表达目的基因。
植物使用包含目的序列(即编码I型DnaJ样多肽或其同源物的核酸或其变体,所述多肽或其同源物在其羧基末端包含CaaX基序)的载体转化。目的序列与一个或多个调控序列(至少与启动子)有效连接。术语“调节元件”、“调控序列”和“启动子”在文中都可以互换使用,且应当取其广义,是指能影响与它们连接的序列表达的调节核酸序列。上述术语包括来源于经典真核生物基因组基因的转录调节序列(包括精确转录起始所需的TATA框,带或不带CCAAT框序列)以及根据发育和/或外界刺激,或以组织特异性方式改变基因表达的另外的调节元件(即上游激活序列、增强子和沉默子)。该术语还包括经典原核基因的转录调节序列,在这种情况下它可以包括-35框序列和/或-10框转录调节序列。术语“调节元件”也包括合成的融合分子或衍生物,其使得核酸分子能够在细胞、组织或器官中表达,激活或增强这种表达。本文所用的术语“有效连接”指启动子序列与目的基因之间的功能性连接,使得启动子序列能起始目的基因的转录。
有利的,任何类型的启动子(无论天然或合成)可以用于驱动核酸序列的表达。优选的,启动子是组织特异性启动子,即能够在某些组织(如叶、根、种子组织等)中优先启动转录的启动子。来自植物的启动子是特别优选的,特别是来自植物的组织特异性启动子。如本文定义的术语“组织特异性”是指在至少一种植物组织或器官中优势表达的启动子,但是由于渗漏启动子表达,其在植物的别处也具有残余表达。更优选的,组织特异性启动子是种子特异性启动子,更特别是分离自编码种子贮藏蛋白质的基因的启动子,特别是胚乳特异性启动子。最优选的,胚乳特异性启动子分离自谷醇溶蛋白基因,如SEQ ID NO:57所代表的稻谷醇溶蛋白RP6(Wen等人,(1993)Plant Physiol 101(3):1115-6)启动子,或类似强度的启动子和/或具有与稻谷醇溶蛋白启动子类似表达模式的启动子。例如,通过将启动子与报道基因结合并且检测报道基因在植物组织中的功能来分析类似强度和/或类似表达模式。一个熟知的报道基因是β-葡糖醛酸糖苷酶,并且比色的GUS染色用来观察植物细胞中的β-葡糖醛酸糖苷酶。要明确本发明的应用不限于SEQ ID NO:1所代表的I型DnaJ样核酸,本发明的应用也不限于当谷醇溶蛋白启动子驱动时I型DnaJ样核酸的表达。
种子特异性启动子的实例示于表4,该启动子或其衍生物可以用于实施本发明方法。要理解下列实例不是穷举的。
表4:用于本发明中的种子特异性启动子的实例
基因来源 | 表达模式 | 参考文献 |
种子-特异性基因 | 种子 | Simon,等人,Plant Mol.Biol.5:191,1985;Scofield,等人,J.Biol.Chem.262:12202,1987.;Baszczynski,等人,Plant Mol.Biol.14:633,1990. |
巴西坚果清蛋白 | 种子 | Pearson,等人,Plant Mol.Biol.18:235-245,1992. |
豆球蛋白 | 种子 | Ellis,等人,Plant Mol.Biol.10:203-214,1988. |
谷蛋白(稻) | 种子 | Takaiwa,等人,Mol.Gen.Genet.208:15-22,1986;Takaiwa,等人,FEBS Letts.221:43-47,1987. |
玉米醇溶蛋白 | 种子 | Matzke等人Plant Mol Biol,14(3):323-32 1990 |
napA | 种子 | Stalberg,等人,Planta 199:515-519,1996. |
小麦LMW和HMW谷蛋白-1 | 胚乳 | Mol Gen Genet 216:81-90,1989;NAR 17:461-2,1989 |
小麦SPA | 种子 | Albani等人,Plant Cell,9:171-184,1997 |
小麦α,β,γ-麦醇溶 | 胚乳 | EMBO 3:1409-15,1984 |
蛋白 | ||
大麦Itr1启动子 | 胚乳 | |
大麦B1,C,D,大麦醇溶蛋白 | 胚乳 | Theor Appl Gen 98:1253-62,1999;Plant J 4:343-55,1993;Mol Gen Genet 250:750-60,1996 |
大麦DOF | 胚乳 | Mena等人,The Plant Journal,116(1):53-62,1998 |
blz2 | 胚乳 | EP99106056.7 |
合成启动子 | 胚乳 | Vicente-Carbajosa等人,Plant J.13:629-640,1998. |
稻谷醇溶蛋白NRP33 | 胚乳 | Wu等人,Plant Cell Physiology 39(8)885-889,1998 |
稻α-球蛋白Glb-1 | 胚乳 | Wu等人,Plant Cell Physiology 39(8)885-889,1998 |
稻OSH1 | 胚 | Sato等人,Proc.Natl.Acad.Sci.美国,93:8117-8122,1996 |
稻α-球蛋白REB/OHP-1 | 胚乳 | Nakase等人.Plant Mol.Biol.33:513-522,1997 |
稻ADP-葡萄糖PP | 胚乳 | Trans Res 6:157-68,1997 |
玉米ESR基因家族 | 胚乳 | Plant J 12:235-46,1997 |
高梁γ-高梁醇溶蛋白 | 胚乳 | PMB 32:1029-35,1996 |
KNOX | 胚 | Postma-Haarsma等人,Plant Mol.Biol.39:257-71,1999 |
稻油质蛋白 | 胚和糊粉 | Wu et at,J.Biochem.,123:386,1998 |
向日葵油质蛋白 | 种子(胚和干种子) | Cummins,等人,Plant Mol.Biol.19:873-876,1992 |
任选的,一个或多个终止子序列也可用在引入植物的构建体中。术语“终止子”包括调控序列,其是位于转录单位末端的DNA序列,标志初级转录物的3′加工和聚腺苷酸化以及转录的终止。其他调节元件可以包括转录增强子以及翻译增强子。本领域技术人员知晓适合用于实施本发明的终止子和增强子序列。此类序列是已知的,或本领域技术人员可以很容易的获得。
本发明的遗传构建体还可以包括在特定细胞类型中维持和/或复制所需要的复制起点序列。一个实例是需要遗传构建体在细菌细胞中作为附加型基因元件(如质粒或黏粒分子)维持的情况。优选的复制起点包括但不局限于fl-ori和colE1。
遗传构建体任选地可包含选择标记基因。本文所用的术语“选择性标记基因”包括赋予表达该基因的细胞以表型,以便于鉴定和/或选择用本发明的核酸构建体转染或转化的细胞的任何基因。合适的标记可选自赋予抗生素或除草剂抗性的标记,其引入新的新陈代谢性状或允许可视选择。选择标记基因的实例包括能赋予抗生素抗性的基因(如使新霉素和卡那霉素磷酸化的nptII,或磷酸化潮霉素的hpt)、赋予除草剂抗性的基因(例如提供Basta抗性的bar;提供草甘膦抗性的aroA或gox)的基因,或提供代谢性状的基因(如允许植物利用甘露糖作为唯一碳源的manA)。可视标记基因导致颜色的形成(例如β-葡糖苷酸酶,GUS)、发光(如荧光素酶)或荧光(绿色荧光蛋白GFP及其衍生物)。
本发明还包括通过本发明方法获得的植物和植物部分。本发明因此提供了可通过本发明方法获得的植物或植物部分,该植物已经在其中引入了编码I型DnaJ样多肽或其同源物的核酸或其变体,该I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序。
本发明还提供了制备在非胁迫条件下生长时具有增加的植物产量的转基因植物的方法,其包括在植物细胞的细胞溶胶中引入和/或表达编码I型DnaJ样多肽或其同源物的核酸或其变体,该I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序。
更具体的,本发明提供了制备具有增加的产量的转基因植物的方法,该方法包括:
(i)在植物、植物部分或植物细胞的细胞溶胶中引入和/或表达编码I型DnaJ样多肽或其同源物的核酸或其变体,该I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序;以及
(ii)在促进植物生长和发育的非胁迫条件下栽培植物、植物部分或植物细胞。
核酸可以直接引入到植物细胞或植物本身中(包括引入到植物的组织、器官或任何其他部分)。根据本发明优选的特征,核酸优选通过转化引入植物中。
本文所涉及的术语“转化”包括把外源多核苷酸向宿主细胞中的转移,而不管转移所用的方法如何。无论是通过器官发生还是胚胎发生能够随后克隆繁殖的植物组织,可以用本发明的遗传构建体转化,并从其再生出整株植物。选择的特定组织将基于可用且最适于待转化的特定物种的克隆繁殖系统而不同。例示性的靶标组织包括叶圆盘、花粉、胚、子叶、下胚轴、雌配子体、胼胝体组织、已存在的分生组织(例如,顶端分生组织,腋芽和根分生组织)以及诱导的分生组织(例如,子叶分生组织和下胚轴分生组织)。多核苷酸可以瞬时或稳定的被引入到宿主细胞中,并可保持非整合状态,例如,作为质粒。可选择的,它可以整合到宿主基因组中。以本领域技术人员所知的方式,之后所得的转化植物细胞可用来再生转化植物。
植物物种的转化目前是一种相当常规的技术。有利的,任何转化方法都可用于将目的基因引入到合适的祖细胞中。转化方法包括使用脂质体、电穿孔、能增加游离DNA摄入的化学品、直接注射DNA到植物中、基因枪颗粒轰击、使用病毒或花粉进行转化以及显微注射。方法可选自用于原生质体的钙/聚乙二醇方法(Krens,FA等人,(1982)Nature 296,72-74;Negrutiu I等人,(1987),Plant Mol.Biol.8,363-373);原生质体的电穿孔(Shillito RD等人,1985 Bio/Technol 3,1099-1102);显微注射到植物材料中(Crossway A.等人,(1986),Mol.Gen.Genet.202,179-185);DNA或RNA包被颗粒轰击(Klein TM等人,(1987),Nature 327,70);用(非整合型)病毒感染等等。优选使用任何用于稻转化的已知方法,通过农杆菌属(Agrobacterium)介导的转化产生表达I型DnaJ样核酸/基因的转基因稻植物,所述的方法例如在任何以下文献中描述的方法:公开的欧洲专利申请EP 1198985 A1,Aldemita和Hodges(Planta 199,612-617,1996);Chan等人(Plant Mol Biol 22(3):491-506,1993),Hiei等人(Plant J.6(2):271-282,1994),其公开的内容以其全文在此引用作为参考。在谷类转化情形下,优选的方法如Ishida等人(Nat Biotechnol 14(6):745-50,1996)或Frame等人(Plant Physiol 129(1):13-22,2002)所述,其公开的内容以其全文在此引用作为参考。
通常在转化以后,选择存在有与目的基因共转移的植物可表达基因所编码的一个或多个标记的植物细胞或细胞群,接着将转化的材料再生成整株植物。
DNA转移和再生之后,可评价推定转化植物中目的基因的存在、拷贝数和/或基因组结构,例如使用Southern分析。可选的或另外的,新引入DNA的表达水平可使用Northern和/或Western分析进行监控,这两种技术都是本领域普通技术人员所熟知的。
产生的转化植物可通过多种方法进行繁殖,如通过无性繁殖或经典育种技术。例如,第一代(或T1)转化植物可以自花授精以产生纯合的第二代(或T2)转化体,并且T2植物可进一步通过经典的育种技术进行繁殖。
产生的转化生物可以是多种形式的。例如,它们可以是转化细胞与非转化细胞的嵌合体;克隆转化体(例如,所有的细胞都被转化以含有表达盒);转化与未转化组织的嫁接体(例如,在植物中,转化的根茎被嫁接到未转化的接穗中)。
本发明显然延及由文中描述的任何方法产生的任何植物细胞或植物,以及所有的植物部分及其繁殖体。本发明还延及囊括由任何上述方法产生的原代转化或转染细胞、组织、器官或整株植物的后代,唯一的要求是该后代表现出与由本发明方法在亲代中产生的基因型和/或表型特征相同的一种或多种基因型和/或表型特征。本发明还包括含有分离的I型DnaJ样核酸或其变体的宿主细胞。根据本发明优选的宿主细胞是植物细胞。本发明还延及植物的可收获部分,例如但不限于种子、叶、果实、花、茎培养物、根茎、块茎和鳞茎。本发明另外涉及(优选直接)来自此类植物的可收获部分的产物,此类产物包括干的颗粒或粉、油、脂肪和脂肪酸、淀粉或蛋白质。
本发明还涵盖I型DnaJ样核酸或其突变体的用途,以及I型DnaJ样多肽或其变体的用途,该I型DnaJ样多肽或其同源物在其羧基末端包含CaaX。一种此类用途涉及增加植物产量,特别是如上所定义的增加产量。
I型DnaJ样核酸或其变体,或I型DnaJ样多肽或其同源物可以用在育种方案中,其中鉴定了与I型DnaJ样基因或其变体遗传连锁的DNA标记。I型DnaJ样核酸/基因或其变体,或I型DnaJ样多肽或其同源物可以用来定义分子标记。之后可以将此DNA或蛋白质标记用在育种方案中以选择具有增加的植物产量的植物。I型DnaJ样基因或其变体可以是例如由表1的SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:5、SEQ ID NO:7、SEQ ID NO:9、SEQ ID NO:11、SEQ ID NO:13、SEQ ID NO:15、SEQID NO:17、SEQ ID NO:19、SEQ ID NO:21、SEQ ID NO:23、SEQ ID NO:25、SEQ ID NO:27、SEQ ID NO:29、SEQ ID NO:31、SEQ ID NO:33、SEQ ID NO:35、SEQ ID NO:37、SEQ ID NO:39、SEQ ID NO:41、SEQID NO:43、SEQ ID NO:45、SEQ ID NO:47、SEQ ID NO:49、SEQ ID NO:51、SEQ ID NO:53或SEQ ID NO:55中任一个所代表的核酸。
还发现I型DnaJ样核酸/基因的等位变体在标记辅助的育种方案中的用途。此类育种方案有时需要通过植物的诱变处理引入等位变异,例如使用EMS诱变;可选的,该方案可以开始于收集无意引起所谓“天然”来源的等位变体。之后通过例如PCR进行等位变体的鉴定。接着是选择步骤,用于选择所讨论序列的优良等位变体,并且其提供植物的增加产量。通常通过监控含有所讨论序列的不同等位变体的植物的产量特性来进行选择,所述的所讨论序列的等位变体如表1的SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:5、SEQ ID NO:7、SEQ ID NO:9、SEQ ID NO:11、SEQ IDNO:13、SEQ ID NO:15、SEQ ID NO:17、SEQ ID NO:19、SEQ ID NO:21、SEQ ID NO:23、SEQ ID NO:25、SEQ ID NO:27、SEQ ID NO:29、SEQ ID NO:31、SEQ ID NO:33、SEQ ID NO:35、SEQ ID NO:37、SEQID NO:39、SEQ ID NO:41、SEQ ID NO:43、SEQ ID NO:45、SEQ ID NO:47、SEQ ID NO:49、SEQ ID NO:51、SEQ ID NO:53或SEQ ID NO:55中任一个的不同等位变体。监控生长特性可以在温室或野外进行。此外任选的步骤包括使已鉴定优良等位变体的植物与另一种植物杂交。例如,这可用于产生目标表型特征的组合。
I型DnaJ样核酸或其变体还可用作探针,以对基因进行遗传和物理作图,这些基因是与之连锁的一部分性状和标志。此类信息可用于植物育种,以开发具有所需表型的品系。I型DnaJ样核酸或其变体的此类用途仅需要长度至少为15个核苷酸的核酸序列。I型DnaJ样核酸或其变体可用作限制性片段长度多态性(RFLP)标记。限制性消化的植物基因组DNA的Southern印迹(Sambrook J,Fritsch EF和Maniatis T(1989)MolecularCloning,A Laboratory ManuaI)可使用I型DnaJ样核酸或其变体进行探测。然后可以使用计算机程序如MapMaker(Lander等人(1987)Genomics1:174-181)对所得的带型进行遗传分析,以构建遗传图谱。此外,该核酸可用于探测Southern印迹,该Southern印迹含有代表亲本和确定的遗传杂交后代的一组个体的限制酶处理的基因组DNA。记录DNA多态性的分离,并用来计算I型DnaJ样核酸或其变体在之前使用该群体获得的遗传图谱中的位置(Botstein等人(1980)Am.J.Hum.Genet.32:314-331)。
在遗传作图中植物基因来源的探针的制备和用途在Bematzky和Tanksley(1986)Plant Mol.Biol.Reporter 4:37-41中描述。众多出版物描述了使用上述方法或其变通形式对特定的cDNA克隆进行遗传作图。例如,F2杂交群体、回交群体、随机交配群体、近亲同基因系及其他的个体组可用于作图。这类方法是本领域技术人员熟知的。
核酸探针也可用于物理作图(即,序列置于物理图上;参见Hoheisel等人,在Non-mammalian Genomic Analysis:A Practical Guide,Academicpress 1996,第319-346页,以及其中引用的参考文献)。
在另一个实施方案中,核酸探针可用于直接荧光原位杂交(FISH)作图(Trask(1991) Trends Genet.7:149-154)。虽然现在的FISH作图方法偏向于使用大的克隆(几kb到几百kb;参见Laan等人(1995)Genome Res.5:13-20),但是灵敏度的提高允许使用较短的探针进行FISH作图。
基于核酸扩增的多种遗传和物理作图方法可以使用所述核酸进行。实例包括等位基因特异性扩增(Kazazian(1989)J.Lab.Clin.Med 11:95-96)、PCR扩增片段的多态性(CAPS;Sheffield等人(1993)Genomics 16:325-332)、等位基因特异性连接(Landegren等人(1988)Science 241:1077-1080)、核苷酸延伸反应(Sokolov(1990)Nucleic Acid Res.18:3671)、放射杂交作图(Walter等人(1997)Nat.Genet.7:22-28)和Happy作图(Dear和Cook(1989)Nucleic Acid Res.17:6795-6807)。为实施这些方法,使用核酸序列设计和制备引物对,以用于扩增反应或引物延伸反应。此类引物的设计是本领域技术人员所熟知的。在采用基于PCR的遗传作图方法中,有必要鉴定跨越了对应于此核酸序列区域作图的亲代之间的DNA序列差异。但是,这对作图方法通常不是必要的。
实施本发明的方法产生如上所述,具有在非胁迫条件下生长的植物中增加植物产量的植物。此增加的植物产量还可与其他经济学上有利的性状组合,如进一步增产性状、对多种胁迫的耐受性、修饰多种结构特征和/或生化和/或生理学特征的性状。
附图说明
本发明现在将参考以下附图进行描述,其中:
图1显示了I型DnaJ样多肽的典型结构域结构。J结构域位于蛋白质的氨基末端并且编码包含高度保守的HPD三肽的HSP70结合结构域。富含甘氨酸和苯丙氨酸的G/F结构域是Hsp陪伴分子活性的特异性靶标蛋白质。四个富含半胱氨酸的结构域参与锌的配位,每个I型DnaJ单体与2个锌离子。CTD结构域是定义的四个结构域中较不保守的,并且可以包含法尼基化基序CaaX。
图2显示了表1的一些I型DnaJ样蛋白质的多重比对,其使用基于改良的ClustalW算法(InforMax,Bethesda,MD,http://www.informaxinc.com)的VNTI AlignX多重对比程序(具有空位开放罚分10以及空位延伸0.05的缺省设置)。J结构域是双下划线的,其四个螺旋是灰色框表示的并且其保守的HPD三肽是框起来的。G/F结构域是用有点的下划线的。两个锌结合结构域I和II以及它们保守的CxxCxGxG是框起来的。在CTD结构域中,法尼基化基序是框起来的。
图3显示了下表中公开的拟南芥I型DnaJ样多肽的比对。J结构域是双下划线的,G/F结构域是黑体下划线的,两个锌结合结构域I和II以及它们保守的CxxCxGxG是框起来的,并且CTD是单下划线的。蛋白质羧基末端的法尼基化基序CaaX当存在时是黑体表示的。在J结构域前的氨基酸序列(由括号分开,大约位置)代表亚细胞导向序列。
MIPS登录号 | NCBI蛋白质登录号 |
At3g44110 | S71199 |
At5g22060 | AAB86799.1 |
At1g28210 | NP849719 |
At1g80030 | AAK60328 |
At2g22360 | AAD22362 |
At3g17830 | NM112664 |
At4g39960 | AAL36077 |
At5g48030 | BAB11067 |
图4显示了用于在稻中表达受到谷醇溶蛋白启动子(内在参考PRO0090)调控的稻I型DnaJ样(内在参考CDS1877)的二元载体。
图5详述了用于实施本发明方法的多核苷酸(从起点到终点)和多肽序列的实例。
实施例
本发明现在将参考以下实施例进行描述,其仅仅是为了例证说明。
DNA操作:除非另外说明,否则重组DNA技术按照(Sambrook(2001)Molecular Cloning:a laboratory manual,第3版,Cold Spring HarborLaboratory Press,CSH,New York)或Ausubel等人(1994),CurrentProtocols in Molecular Biology,Current Protocols的卷1和2中描述的标准方案进行。用于植物分子工作的标准材料和方法描述于R.D.D Croy的Plant Molecular Biology Labfase(1993),由BIOS Scientific PublicationsLtd(UK)和Blackwell Scientific Publications(UK)出版。
实施例1:基因克隆
使用稻幼苗cDNA文库(Invitrogen,Paisley,UK)作为模板,PCR扩增稻I型DnaJ样基因(CDS1877)。反转录从幼苗提取的RNA后,把cDNA克隆到pCMV Sport 6.0中。文库的平均插入大小为1.6 kb,并且克隆的原始数为1.67×107 cfu。原始滴度确定为3.34×106 cfu/ml,并且在第一轮扩增以后变成6×1010 cfu/ml。质粒提取以后,200ng的模板用于50μlPCR混合物中。包含用于Gateway重组的AttB位点的引物prm04266(SEQ IDNO:58;有义,起始密码子为黑体,AttB1位点是斜体:5’GGGGACAAGTTTGTACAAAAAAGCAGGCTTCACAATGTACGGACGCATGCC 3’)和prm04267(SEQ ID NO:59;反向互补,终止密码子是黑体,AttB2位点为斜体:5’GGGGACCACTTTGTACAAGAAAGCTGGGTGCATCGAATTGTTCTTACTGC 3’)用于PCR扩增。使用Hifi Tag DNA聚合酶,在标准条件下进行PCR。同样使用标准方法扩增和纯化1340 bp的PCR片段(包括attB位点)。然后进行Gateway方法的第一步,即BP反应,在此期间,PCR片段与pDONR201质粒体内重组以产生“进入克隆(entry clone)”p04452(根据Gateway术语)。质粒pDONR201作为Gatewaytechnology的一部分购自Invitrogen。
实施例2:载体构建
继之在LR反应中使用进入克隆p04452和用于稻转化的指定载体p00830。该载体包含位于T-DNA边界内的以下部分作为功能性元件:植物选择性标记;可筛选标记表达盒;旨在与已克隆到进入克隆中的目的序列进行LR体内重组的Gateway盒。用于胚乳特异性表达的稻RP6谷醇溶蛋白启动子(PRO0090)(SEQ ID NO:57;Wen等人,(1993)Plant Physiol101(3):1115-6)位于该Gateway盒的上游。
LR重组步骤之后,将得到的表达载体p072(图4)转化到农杆菌属菌株LBA4404中,并且随后转化稻植株。使转化的稻植株生长,然后检验实施例3中所述的参数。
实施例3:受到稻RP6启动子调控的I型DnaJ样的评估和结果
产生了大约15到20个独立的T0稻转化体。将初级转化体从组织培养箱转移到温室中生长,并收获T1种子。5个事件得以保留,其中T1后代发生3∶1的转基因存在/缺乏分离。对于这些事件中的每一个,通过监控可视标记表达,选择了大约10个包含转基因的T1幼苗(杂合子和纯合子),以及大约10个缺乏转基因的T1幼苗(失效合子)。将4个T1事件在T2代中使用与T1代相同的评价方法进行进一步评估,但是不必是每个事件更多个体。
统计学分析:F-检验
两因子的ANOVA(可变性分析)用作统计模型,用于植物表型特征的综合评价。对用本发明的基因转化的所有事件的所有植株的所有测量参数进行F检验。进行F检验以检查基因对所有转化事件的作用,并检验基因的总效应,亦称之为整体基因效应。真实整体基因效应的显著性阈值设置为F检验的5%概率水平。显著的F检验值显示基因效应,意味着不仅仅是基因的存在或位置引起表型的差异。
由于进行了具有重叠事件的两个实验,除分析上述外还进行组合分析。这可用于检查两个实验中作用的一致性,如果在这种情形下,其可用于从两个实验中收集证据,则增加结论的可信性。使用的方法是考虑到数据的多级结构(即实验-事件-分离子)的混合模型方法。通过对比卡方分布的似然比值检验得到p值。
3.1种子相关的参数测量
收获成熟的初级圆锥花序、装袋并贴上条码标签,然后在37℃烤箱中干燥三天。随后将圆锥花序脱粒,收集所有的种子并且计数。使用鼓风装置将饱满外壳和空外壳分开。丢弃空外壳,并对保留的部份再次计数。在分析天平上称量饱满的外壳。通过计数分离步骤之后保留的饱满外壳的数目确定饱满的种子数。通过称量从植物收获的所有饱满外壳来测量总种子产量。每株植株的总种子数是通过计数收获自植物的外壳数来测量的。将本发明的收获指数定义为总种子产量和地上面积(mm2)的比乘以因子106。
3.2地上面积
植物地上面积通过计数排除背景后地上植物部分图像的像素总数测定。这个值为在同一时间点从不同角度拍摄的图像的平均数,并通过校准将其转化为以平方毫米表示的物理表面值。实验显示,这种方式测定的地上植物面积与植物的生物量相关。
下列结果的表(表5)显示了转基因与相应的失效合子的收获指数之间的百分比差异。
进行组合分析证实了两个实验作用的一致性,并且因此增加了结论的可信性。
表5:收获指数
收获指数 | |||
T1%增加 | T2%增加 | 组合的P值 | |
事件1 | 55 | 24 | 0.0024 |
事件2 | 35 | 11 | 0.0585 |
总共 | 9(5个事件) | 8(4个事件) | 0.0063 |
序列表
<110>克罗普迪塞恩股份有限公司
<120>产量增加的植物及其制备方法
<130>CD-128-PCT
<150>EP 04106985.7
<151>2004-12-24
<150>US 60/641,688
<151>2005-01-06
<160>59
<170>PatentIn版本3.3
<210>1
<211>1263
<212>DNA
<213>稻(Oryza sativa)
<400>1
atgtacggac gcatgccaaa gaagagtaac aataccaagt attatgaggt gcttggtgta 60
tctaagacag caacccagga tgagctgaag aaagcgtacc gtaaagctgc cattaaaaac 120
caccctgata agggtggaga ccctgagaag tttaaagaat tggctcaagc ttacgaggtt 180
cttaatgatc ctgaaaagag ggaaatctat gaccaatatg gcgaggatgc actcaaagaa 240
ggaatgggag gaggcagcag cagtgatttc catagtccct tcgatttatt tgagcaaatt 300
tttcagaatc gtggtggctt tgggggtaga ggacacagac aaaagcgtgg cgaagatgtg 360
gtacatacta tgaaggtttc tttagaagac ctgtataatg gtactaccaa aaaactgtct 420
ttgtcacgga atgctctgtg cacaaagtgc aagggtaaag gatccaagag tggggcagca 480
gcaacttgcc atggttgtca tggtgcagga atgagaacaa taacaagaca aattgggctt 540
ggcatgatcc aacagatgaa cactgtttgc cctgaatgca gaggatcagg tgagatgata 600
agtgacaagg ataaatgccc gagttgtaag ggaaacaaag tagtccagca gaagaaggtc 660
ttggaggttc atgttgagaa gggaatgcaa catggccaaa agattgtatt ccagggtgaa 720
gctgatgaag ctcctgatac agtgacagga gacatagttt ttgtcttgca acttaaagac 780
cacccaaaat ttaagaggaa gtttgatgac ctctttactg agcacacaat ctccctgacc 840
gaggctctgt gtggcttcca gtttgttcta acccatcttg atggtcggca actcctaatc 900
aaatctaatc caggggaggt tataaaacct ggtcaacaca aggccatcaa tgatgaaggc 960
atgccccagc atggccgccc tttcatgaaa ggtcgtcttt ttgttgaatt caacgtggag 1020
tttcctgagc ctggtgcact cactcctggc caatgccgat cgcttgagaa gattttgcca 1080
ccacgaccca ggaatcaatt gtcagacatg gagctagatc aatgtgagga gaccaccatg 1140
catgatgtca acatagaaga ggagatgagg cgcaggcagc agcacaggcg gcaggaagca 1200
tatgatgaag acgacgacga ggatgctgga gctggaccaa gggtacagtg tgcccagcag 1260
taa 1263
<210>2
<211>420
<212>PRT
<213>稻
<400>2
Met Tyr Gly Arg Met Pro Lys Lys Ser Asn Asn Thr Lys Tyr Tyr Glu
1 5 10 15
Val Leu Gly Val Ser Lys Thr Ala Thr Gln Asp Glu Leu Lys Lys Ala
20 25 30
Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Asn Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Gly Gly Ser Ser Ser Asp Phe His Ser Pro Phe Asp Leu
85 90 95
Phe Glu Gln Ile Phe Gln Asn Arg Gly Gly Phe Gly Gly Arg Gly His
100 105 110
Arg Gln Lys Arg Gly Glu Asp Val Val His Thr Met Lys Val Ser Leu
115 120 125
Glu Asp Leu Tyr Asn Gly Thr Thr Lys Lys Leu Ser Leu Ser Arg Asn
130 135 140
Ala Leu Cys Thr Lys Cys Lys Gly Lys Gly Ser Lys Ser Gly Ala Ala
145 150 155 160
Ala Thr Cys His Gly Cys His Gly Ala Gly Met Arg Thr Ile Thr Arg
165 170 175
Gln Ile Gly Leu Gly Met Ile Gln Gln Met Asn Thr Val Cys Pro Glu
180 185 190
Cys Arg Gly Ser Gly Glu Met Ile Ser Asp Lys Asp Lys Cys Pro Ser
195 200 205
Cys Lys Gly Asn Lys Val Val Gln Gln Lys Lys Val Leu Glu Val His
210 215 220
Val Glu Lys Gly Met Gln His Gly Gln Lys Ile Val Phe Gln Gly Glu
225 230 235 240
Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Val Phe Val Leu
245 250 255
Gln Leu Lys Asp His Pro Lys Phe Lys Arg Lys Phe Asp Asp Leu Phe
260 265 270
Thr Glu His Thr Ile Ser Leu Thr Glu Ala Leu Cys Gly Phe Gln Phe
275 280 285
Val Leu Thr His Leu Asp Gly Arg Gln Leu Leu Ile Lys Ser Asn Pro
290 295 300
Gly Glu Val Ile Lys Pro Gly Gln His Lys Ala Ile Asn Asp Glu Gly
305 310 315 320
Met Pro Gln His Gly Arg Pro Phe Met Lys Gly Arg Leu Phe Val Glu
325 330 335
Phe Asn Val Glu Phe Pro Glu Pro Gly Ala Leu Thr Pro Gly Gln Cys
340 345 350
Arg Ser Leu Glu Lys Ile Leu Pro Pro Arg Pro Arg Asn Gln Leu Ser
355 360 365
Asp Met Glu Leu Asp Gln Cys Glu Glu Thr Thr Met His Asp Val Asn
370 375 380
Ile Glu Glu Glu Met Arg Arg Arg Gln Gln His Arg Arg Gln Glu Ala
385 390 395 400
Tyr Asp Glu Asp Asp Asp Glu Asp Ala Gly Ala Gly Pro Arg Val Gln
405 410 415
Cys Ala Gln Gln
420
<210>3
<211>1251
<212>DNA
<213>稻
<400>3
atgtttgggc gtgtaccgag gagtaacaac accaagtact atgaggttct tggagttcct 60
aaaactgcaa gcaaggatga gctaaagaag gcataccgga aggctgccat aaaaaaccat 120
cctgacaagg gaggggatcc agagaagttt aaagaattat cacaagcgta tgaggttctc 180
actgatcctg agaagagaga catatatgac caatatgggg aggatgctct taaggatgga 240
atgggaggag gcagtgactt ccataatcca tttgacatat ttgagcagtt tttcgggggt 300
ggtgcctttg gggggagtag ctcaagagta cgcagacaga gacgtggtga agatgtggcg 360
catactttga aggtgtcttt agaagatgtg tataatggat ctatgaagaa actatcatta 420
tcacgaaata ttctgtgccc aaagtgcaaa ggaaaaggga ccaaatctga ggctccagca 480
acatgctatg gttgtcatgg tgtaggaatg aggaatataa tgcgacagat aggactaggc 540
atgattcaac atatgcagac tgtctgtcct gaatgcagag gatcaggtga gatcataagt 600
gacagggata aatgcacaaa ctgcagagct agcaaagtta ttcaggagaa aaaggtgctt 660
gaggttcata ttgagaaggg aatgcaacat ggccaaaaaa ttgtattcca aggtgaagct 720
gatgaagctc ctgatacagt gacaggagat atagtattta tcttgcaagt taaggtacat 780
ccaagattta agaggaaata tgatgacctg ttcattgagc gcacaatctc tttaactgag 840
gcattgtgtg ggttccaatt catcctcact catctggaca gtaggcagct cctaatcaag 900
gcaaatcctg gcgaaattat taaacctggt caacacaagg ccataaatga tgagggaatg 960
ccacaccatg gccggccttt catgaagggc cgtctctttg tggaattcaa tgttgagttc 1020
cctgaatctg gtgtactctc ccgtgaccaa tgccgggcac ttgagatgat cctaccacct 1080
aaacctgggc accaattatc agatatggac ctggatcaat gtgaggaaac taccatgcat 1140
gatgtgaaca tagaagagga gatgaggcgc aagcagtatc aaaggaagca ggaagcgtac 1200
gacgaagatg aggaggagga tgctccaaga gtacagtgtg ctcaacagta a 1251
<210>4
<211>416
<212>PRT
<213>稻
<400>4
Met Phe Gly Arg Val Pro Arg Ser Asn Asn Thr Lys Tyr Tyr Glu Val
1 5 10 15
Leu Gly Val Pro Lys Thr Ala Ser Lys Asp Glu Leu Lys Lys Ala Tyr
20 25 30
Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro Glu
35 40 45
Lys Phe Lys Glu Leu Ser Gln Ala Tyr Glu Val Leu Thr Asp Pro Glu
50 55 60
Lys Arg Asp Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Asp Gly
65 70 75 80
Met Gly Gly Gly Ser Asp Phe His Asn Pro Phe Asp Ile Phe Glu Gln
85 90 95
Phe Phe Gly Gly Gly Ala Phe Gly Gly Ser Ser Ser Arg Val Arg Arg
100 105 110
Gln Arg Arg Gly Glu Asp Val Ala His Thr Leu Lys Val Ser Leu Glu
115 120 125
Asp Val Tyr Asn Gly Ser Met Lys Lys Leu Ser Leu Ser Arg Asn Ile
130 135 140
Leu Cys Pro Lys Cys Lys Gly Lys Gly Thr Lys Ser Glu Ala Pro Ala
145 150 155 160
Thr Cys Tyr Gly Cys His Gly Val Gly Met Arg Asn Ile Met Arg Gln
165 170 175
Ile Gly Leu Gly Met Ile Gln His Met Gln Thr Val Cys Pro Glu Cys
180 185 190
Arg Gly Ser Gly Glu Ile Ile Ser Asp Arg Asp Lys Cys Thr Asn Cys
195 200 205
Arg Ala Ser Lys Val Ile Gln Glu Lys Lys Val Leu Glu Val His Ile
210 215 220
Glu Lys Gly Met Gln His Gly Gln Lys Ile Val Phe Gln Gly Glu Ala
225 230 235 240
Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Val Phe Ile Leu Gln
245 250 255
Val Lys Val His Pro Arg Phe Lys Arg Lys Tyr Asp Asp Leu Phe Ile
260 265 270
Glu Arg Thr Ile Ser Leu Thr Glu Ala Leu Cys Gly Phe Gln Phe Ile
275 280 285
Leu Thr His Leu Asp Ser Arg Gln Leu Leu Ile Lys Ala Asn Pro Gly
290 295 300
Glu Ile Ile Lys Pro Gly Gln His Lys Ala Ile Asn Asp Glu Gly Met
305 310 315 320
Pro His His Gly Arg Pro Phe Met Lys Gly Arg Leu Phe Val Glu Phe
325 330 335
Asn Val Glu Phe Pro Glu Ser Gly Val Leu Ser Arg Asp Gln Cys Arg
340 345 350
Ala Leu Glu Met Ile Leu Pro Pro Lys Pro Gly His Gln Leu Ser Asp
355 360 365
Met Asp Leu Asp Gln Cys Glu Glu Thr Thr Met His Asp Val Asn Ile
370 375 380
Glu Glu Glu Met Arg Arg Lys Gln Tyr Gln Arg Lys Gln Glu Ala Tyr
385 390 395 400
Asp Glu Asp Glu Glu Glu Asp Ala Pro Arg Val Gln Cys Ala Gln Gln
405 410 415
<210>5
<211>1254
<212>DNA
<213>稻
<400>5
atgttcgggc gcgcgccgaa gaagagcgac aacaccaagt actacgagat cctgggggtc 60
cccaagaccg cctcccagga cgacctcaag aaggcgtacc gcaaggccgc catcaagaac 120
caccccgaca agggcggcga ccccgagaag ttcaaggagc ttgcacaagc ttatgaggta 180
ttgagtgacc cggagaaacg tgaaatctat gaccaatatg gtgaagatgc cctcaaggaa 240
ggaatgggtg gaggcggatc ccatgttgat ccatttgaca tcttttcatc attctttgga 300
ccttcttttg gtggtggtgg cagcagcagg ggcagaaggc aaaggagggg agaggatgtg 360
atccatccgc ttaaggtttc tctagaagat ctttacaatg gtacttcaaa gaagctctct 420
ctttcccgca atgtcctctg cgccaagtgc aagggcaagg gttccaagtc tggtgcttcc 480
atgaggtgcc caggttgcca ggggtctggc atgaaaatca ccatccgcca gctggggccc 540
tccatgatac agcagatgca gcagccttgc aatgagtgta aggggactgg agagagcatt 600
aatgagaagg atcgctgccc aggctgcaag ggcgagaagg ttattcagga gaagaaggtt 660
ctggaggttc acgttgagaa ggggatgcaa cacaatcaga agatcacttt ccctggtgaa 720
gctgatgagg cgcctgatac cgttacggga gacattgtat tcgtcctcca gcagaaggac 780
cactccaagt tcaaaaggaa gggcgatgat ctcttttatg agcacacctt atctctgact 840
gaagcacttt gtggtttcca atttgtcctg acacatctgg acaacagaca gctgctcatt 900
aagtcaaacc ccggtgaagt tgttaagcct gaccaattca aggcaataaa cgatgaggga 960
atgccaatgt accagaggcc tttcatgaag gggaagctct acattcattt cacggtggag 1020
ttccctgatt ccctggcgcc tgaacaatgc aaggctctcg aggctgtgct tccaccgaag 1080
cctgcatccc agctgacaga aatggagata gatgaatgcg aggagaccac gatgcacgat 1140
gtcaacaaca ttgaggaaga gatgcgcagg aaagcccaag ctgctcagga ggcgtatgat 1200
gaggacgatg agatgcctgg aggtgcccag agagttcagt gcgcgcaaca gtaa 1254
<210>6
<211>417
<212>PRT
<213>稻
<400>6
Met Phe Gly Arg Ala Pro Lys Lys Ser Asp Asn Thr Lys Tyr Tyr Glu
1 5 10 15
Ile Leu Gly Val Pro Lys Thr Ala Ser Gln Asp Asp Leu Lys Lys Ala
20 25 30
Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Gly Gly Gly Ser His Val Asp Pro Phe Asp Ile Phe Ser
85 90 95
Ser Phe Phe Gly Pro Ser Phe Gly Gly Gly Gly Ser Ser Arg Gly Arg
100 105 110
Arg Gln Arg Arg Gly Glu Asp Val Ile His Pro Leu Lys Val Ser Leu
115 120 125
Glu Asp Leu Tyr Asn Gly Thr Ser Lys Lys Leu Ser Leu Ser Arg Asn
130 135 140
Val Leu Cys Ala Lys Cys Lys Gly Lys Gly Ser Lys Ser Gly Ala Ser
145 150 155 160
Met Arg Cys Pro Gly Cys Gln Gly Ser Gly Met Lys Ile Thr Ile Arg
165 170 175
Gln Leu Gly Pro Ser Met Ile Gln Gln Met Gln Gln Pro Cys Asn Glu
180 185 190
Cys Lys Gly Thr Gly Glu Ser Ile Asn Glu Lys Asp Arg Cys Pro Gly
195 200 205
Cys Lys Gly Glu Lys Val Ile Gln Glu Lys Lys Val Leu Glu Val His
210 215 220
Val Glu Lys Gly Met Gln His Asn Gln Lys Ile Thr Phe Pro Gly Glu
225 230 235 240
Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Val Phe Val Leu
245 250 255
Gln Gln Lys Asp His Ser Lys Phe Lys Arg Lys Gly Asp Asp Leu Phe
260 265 270
Tyr Glu His Thr Leu Ser Leu Thr Glu Ala Leu Cys Gly Phe Gln Phe
275 280 285
Val Leu Thr His Leu Asp Asn Arg Gln Leu Leu Ile Lys Ser Asn Pro
290 295 300
Gly Glu Val Val Lys Pro Asp Gln Phe Lys Ala Ile Asn Asp Glu Gly
305 310 315 320
Met Pro Met Tyr Gln Arg Pro Phe Met Lys Gly Lys Leu Tyr Ile His
325 330 335
Phe Thr Val Glu Phe Pro Asp Ser Leu Ala Pro Glu Gln Cys Lys Ala
340 345 350
Leu Glu Ala Val Leu Pro Pro Lys Pro Ala Ser Gln Leu Thr Glu Met
355 360 365
Glu Ile Asp Glu Cys Glu Glu Thr Thr Met His Asp Val Asn Asn Ile
370 375 380
Glu Glu Glu Met Arg Arg Lys Ala Gln Ala Ala Gln Glu Ala Tyr Asp
385 390 395 400
Glu Asp Asp Glu Met Pro Gly Gly Ala Gln Arg Val Gln Cys Ala Gln
405 410 415
Gln
<210>7
<211>1254
<212>DNA
<213>稻
<400>7
atgttcgggc gcgcgccgaa gaagagcgac aacacgcggt actacgaggt gcttggggtg 60
cccaaggatg cgtcccagga tgacctcaag aaggcgtacc gcaaggccgc catcaagaac 120
caccccgaca agggcggaga ccccgagaag ttcaaggaat tggctcaggc ttatgaagtc 180
ctgagtgacc ctgagaagcg tgaaatctat gatcagtacg gtgaagatgc tctcaaggag 240
gggatgggtc ctggtggtgg gatgcatgac ccatttgaca ttttttcctc attctttgga 300
ggtggctttg gaggtggtag cagtaggggc aggagacagc gtaggggaga ggatgtggtt 360
caccctctga aggtttctct ggaggaattg tacaatggca catcaaagaa gctctccctt 420
tctcgcaatg tgctctgctc caagtgcaat ggcaagggct cgaaatctgg tgcttccatg 480
aagtgctctg gttgtcaagg ttctggtatg aaggtccaaa ttcgccagtt ggggccagga 540
atgattcagc aaatgcaaca tccctgcaat gagtgcaagg gaactggtga gaccatcagc 600
gacaaggata gatgcccagg ctgcaagggt gagaaggtgg cgcaggagaa gaaggttctt 660
gaggtggtgg tcgagaaggg catgcagaat ggacagaaga tcaccttccc tggtgaggct 720
gatgaagcgc ccgatactgt cactggagac attatcttcg tcctccagca gaaggagcat 780
cccaagttca agagaaaggg agatgacctc ttctacgagc acaccctgaa cctcactgag 840
gccctttgtg gcttccagtt tgttctcact cacttggaca acaggcagct gcttatcaag 900
tccaagcccg gtgaagttgt caagcctgat tcattcaagg ctgtcaacga cgagggcatg 960
ccgatgtacc agcggccatt catgaagggg aagctctaca tccacttctc cgtggaattc 1020
cccgactctt tgaaccctga ccagtgcaag gccctggaga ccgtcctccc gccaaggccg 1080
gtgtcgcagt acaccgacat ggagctcgac gagtgcgagg agaccatgcc gtacgacgtg 1140
aacatcgagg aggagatgag gaggcggcag caacagcagc agcaggaggc atacgacgag 1200
gacgaggaca tgcacggcgg cggcgcccag cgcgtgcagt gcgcgcagca gtaa 1254
<210>8
<211>417
<212>PRT
<213>稻
<400>8
Met Phe Gly Arg Ala Pro Lys Lys Ser Asp Asn Thr Arg Tyr Tyr Glu
1 5 10 15
Val Leu Gly Val Pro Lys Asp Ala Ser Gln Asp Asp Leu Lys Lys Ala
20 25 30
Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Pro Gly Gly Gly Met His Asp Pro Phe Asp Ile Phe Ser
85 90 95
Ser Phe Phe Gly Gly Gly Phe Gly Gly Gly Ser Ser Arg Gly Arg Arg
100 105 110
Gln Arg Arg Gly Glu Asp Val Val His Pro Leu Lys Val Ser Leu Glu
115 120 125
Glu Leu Tyr Asn Gly Thr Ser Lys Lys Leu Ser Leu Ser Arg Asn Val
l30 135 140
Leu Cys Ser Lys Cys Asn Gly Lys Gly Ser Lys Ser Gly Ala Ser Met
145 150 155 160
Lys Cys Ser Gly Cys Gln Gly Ser Gly Met Lys Val Gln Ile Arg Gln
165 170 175
Leu Gly Pro Gly Met Ile Gln Gln Met Gln His Pro Cys Asn Glu Cys
180 185 190
Lys Gly Thr Gly Glu Thr Ile Ser Asp Lys Asp Arg Cys Pro Gly Cys
195 200 205
Lys Gly Glu Lys Val Ala Gln Glu Lys Lys Val Leu Glu Val Val Val
210 215 220
Glu Lys Gly Met Gln Asn Gly Gln Lys Ile Thr Phe Pro Gly Glu Ala
225 230 235 240
lu Ala Pro Asp Thr Val Thr Gly Asp Ile Ile Phe Val Leu Gln
245 250 255
Gln Lys Glu His Pro Lys Phe Lys Arg Lys Gly Asp Asp Leu Phe Tyr
260 265 270
Glu His Thr Leu Asn Leu Thr Glu Ala Leu Cys Gly Phe Gln Phe Val
275 280 285
Leu Thr His Leu Asp Asn Arg Gln Leu Leu Ile Lys Ser Lys Pro Gly
290 295 300
Glu Val Val Lys Pro Asp Ser Phe Lys Ala Val Asn Asp Glu Gly Met
305 310 315 320
Pro Met Tyr Gln Arg Pro Phe Met Lys Gly Lys Leu Tyr Ile His Phe
325 330 335
Ser Val Glu Phe Pro Asp Ser Leu Asn Pro Asp Gln Cys Lys Ala Leu
340 345 350
Glu Thr Val Leu Pro Pro Arg Pro Val Ser Gln Tyr Thr Asp Met Glu
355 360 365
Leu Asp Glu Cys Glu Glu Thr Met Pro Tyr Asp Val Asn Ile Glu Glu
370 375 380
Glu Met Arg Arg Arg Gln Gln Gln Gln Gln Gln Glu Ala Tyr Asp Glu
385 390 395 400
Asp Glu Asp Met His Gly Gly Gly Ala Gln Arg Val Gln Cys Ala Gln
405 410 415
Gln
<210>9
<211>1260
<212>DNA
<213>玉米(Zea mays)
<400>9
atgttcgggc gcgcgccgaa gaagagcgac aacaccaagt actacgagat cctcggggtg 60
cccaagtcgg cgtcccagga cgatctcaag aaggcctacc gcaaggctgc tatcaagaac 120
caccccgaca agggcggtga ccccgagaag ttcaaggagc tcgcacaagc ctatgaggtt 180
ttgagtgatc cagagaaacg tgagatttat gatcagtatg gtgaagatgc ccttaaggaa 240
ggaatgggcg gtggaggatc ccatgttgat ccatttgaca tcttctcatc attttttgga 300
ccctcttttg gaggaggtgg tggaagcagc aggggaagaa ggcaaaggag gggagaagat 360
gtagttcacc cacttaaagt ttctctggaa gatctttaca atggcacctc aaagaagctc 420
tctctttcgc gcaatgtcat ctgctccaag tgcaagggca agggctcgaa gtctggtgcc 480
tcaatgaggt gccctggttg ccagggctca ggcatgaaag tcactattcg tcagctgggc 540
ccttccatga tacagcagat gcagcagcct tgcaatgagt gcaaggggac tggagagagc 600
atcaatgaga aggaccgctg tccagggtgc aagggtgaga aggtcattca agagaagaaa 660
gttcttgagg ttcatgttga gaaggggatg caacacaacc agaagatcac cttccctggt 720
gaagctgatg aagcgcctga tactgtcact ggagacattg tattcgtcct ccaacagaag 780
gatcactcca aattcaaaag aaagggtgaa gatctgttct atgagcacac cttgtctctg 840
accgaagcac tatgtgggtt ccaatttgtt cttacacatc tggacaacag gcagcttctc 900
atcaaatcag accctggtga agttgttaaa cctgaccaat tcaaggcgat taatgatgag 960
gggatgccaa tttaccagag gcctttcatg aaggggaagc tgtacatcca tttcacggtg 1020
gagttccctg actcgttggc accagagcag tgcaaggctc tcgagacagt acttccacca 1080
aggccttcat ccaagctgac agacatggag atagatgaat gcgaggagac gactatgcat 1140
gatgtgaaca acatcgagga agagatgcgc aagaagcaag ctcacgctgc ccaggaggcg 1200
tacgaggagg acgacgagat gccgggcgga gcccagagag tgcagtgcgc gcagcagtaa 1260
<210>10
<211>419
<212>PRT
<213>玉米
<400>10
Met Phe Gly Arg Ala Pro Lys Lys Ser Asp Asn Thr Lys Tyr Tyr Glu
1 5 10 15
Ile Leu Gly Val Pro Lys Ser Ala Ser Gln Asp Asp Leu Lys Lys Ala
20 25 30
Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Gly Gly Gly Ser His Val Asp Pro Phe Asp Ile Phe Ser
85 90 95
Ser Phe Phe Gly Pro Ser Phe Gly Gly Gly Gly Gly Ser Ser Arg Gly
100 105 110
Arg Arg Gln Arg Arg Gly Glu Asp Val Val His Pro Leu Lys Val Ser
115 120 125
Leu Glu Asp Leu Tyr Asn Gly Thr Ser Lys Lys Leu Ser Leu Ser Arg
130 135 140
Asn Val Ile Cys Ser Lys Cys Lys Gly Lys Gly Ser Lys Ser Gly Ala
145 150 155 160
Ser Met Arg Cys Pro Gly Cys Gln Gly Ser Gly Met Lys Val Thr Ile
165 170 175
Arg Gln Leu Gly Pro Ser Met Ile Gln Gln Met Gln Gln Pro Cys Asn
180 185 190
Glu Cys Lys Gly Thr Gly Glu Ser Ile Asn Glu Lys Asp Arg Cys Pro
195 200 205
Gly Cys Lys Gly Glu Lys Val Ile Gln Glu Lys Lys Val Leu Glu Val
210 215 220
His Val Glu Lys Gly Met Gln His Asn Gln Lys Ile Thr Phe Pro Gly
225 230 235 240
Glu Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Val Phe Val
245 250 255
Leu Gln Gln Lys Asp His Ser Lys Phe Lys Arg Lys Gly Glu Asp Leu
260 265 270
Phe Tyr Glu His Thr Leu Ser Leu Thr Glu Ala Leu Cys Gly Phe Gln
275 280 285
Phe Val Leu Thr His Leu Asp Asn Arg Gln Leu Leu Ile Lys Ser Asp
290 295 300
Pro Gly Glu Val Val Lys Pro Asp Gln Phe Lys Ala Ile Asn Asp Glu
305 310 315 320
Gly Met Pro Ile Tyr Gln Arg Pro Phe Met Lys Gly Lys Leu Tyr Ile
325 330 335
His Phe Thr Val Glu Phe Pro Asp Ser Leu Ala Pro Glu Gln Cys Lys
340 345 350
Ala Leu Glu Thr Val Leu Pro Pro Arg Pro Ser Ser Lys Leu Thr Asp
355 360 365
Met Glu Ile Asp Glu Cys Glu Glu Thr Thr Met His Asp Val Asn Asn
370 375 380
Ile Glu Glu Glu Met Arg Arg Lys Gln Ala His Ala Ala Gln Glu Ala
385 390 395 400
Tyr Glu Glu Asp Asp Glu Met Pro Gly Gly Ala Gln Arg Val Gln Cys
405 410 415
Ala Gln Gln
<210>11
<211>1257
<212>DNA
<213>玉米
<400>11
atgttcgggc gcgcgccgaa gaagagcgac aacacacggt actacgagat cctcggggtc 60
tccaaggacg cgtcccagga tgacctcaag aaagcctacc gcaaggccgc catcaagaac 120
caccccgaca agggcggcga tcccgagaag ttcaaggagc tagctcaggc ttatgaggtc 180
ctcagtgatc ctgaaaagcg ggagatttat gatcaatatg gtgaggatgc cctcaaggag 240
ggaatgggag gtggtggagg gatgcacgat ccctttgaca tattccagtc attctttggt 300
ggtggaagcc cttttggagg tggtggcagc agtaggggca gaaggcagcg aaggggagag 360
gatgtggttc atcctctaaa ggtttctctg gaggatttgt acaatggcac atcaaagaag 420
ctctctctgt cccgcagtgt cctctgctcc aagtgcaatg gtaagggttc aaagtctgga 480
gcttcatcga ggtgtgctgg ttgccaaggt tctggcttta aggtccaaat ccggcagttg 540
tggaa tgatccagca aatgcagcat ccttgcaacg agtgcaaggg ttctggagag 600
acaatcagcg acaaggatag atgcccacag tgcaagggtg ataaagttgt gcaggagaag 660
aaggttcttg aagtgtttgt ggagaaaggc atgcagaatg ggcagaagat cacattccct 720
ggtgaagctg atgaagcgcc tgacactgtc actggagata tcatttttgt tctccagcag 780
aaggagcatc ccaagttcaa gagaaagggc gatgacctct tctacgagca caccctgacc 840
ttgactgaat ctctgtgtgg cttccagttt gttgtgactc acttggataa caggcagctg 900
ctgatcaaat caaatccggg cgaagttgtg aagcctgatt ctttcaaggc gatcaacgac 960
gaaggcatgc ccatgtacca gaggccgttc atgaagggca agctgtacat ccacttctcg 1020
gtggagttcc cggactcgct gagcccggag cagtgcaagg ccctggaggc tgtgctcccg 1080
cccaagccgg tgtcgcagta caccgacatg gagctggacg agtgcgagga gacgatgccc 1140
tatgacgtga acatcgaagc ggagatgcgg aggcggcagc agcagcacca ggaggcctac 1200
gacgaggatg aggacatgcc gggcggcgcg cagagggtgc agtgcgccca gcagtag 1257
<210>12
<211>418
<212>PRT
<213>玉米
<400>12
Met Phe Gly Arg Ala Pro Lys Lys Ser Asp Asn Thr Arg Tyr Tyr Glu
1 5 10 15
Ile Leu Gly Val Ser Lys Asp Ala Ser Gln Asp Asp Leu Lys Lys Ala
20 25 30
Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Gly Gly Gly Gly Met His Asp Pro Phe Asp Ile Phe Gln
85 90 95
Ser Phe Phe Gly Gly Gly Ser Pro Phe Gly Gly Gly Gly Ser Ser Arg
100 105 110
Gly Arg Arg Gln Arg Arg Gly Glu Asp Val Val His Pro Leu Lys Val
115 120 125
Ser Leu Glu Asp Leu Tyr Asn Gly Thr Ser Lys Lys Leu Ser Leu Ser
130 135 140
Arg Ser Val Leu Cys Ser Lys Cys Asn Gly Lys Gly Ser Lys Ser Gly
145 150 155 160
Ala Ser Ser Arg Cys Ala Gly Cys Gln Gly Ser Gly Phe Lys Val Gln
165 170 175
Ile Arg Gln Leu Gly Pro Gly Met Ile Gln Gln Met Gln His Pro Cys
180 185 190
Asn Glu Cys Lys Gly Ser Gly Glu Thr Ile Ser Asp Lys Asp Arg Cys
195 200 205
Pro Gln Cys Lys Gly Asp Lys Val Val Gln Glu Lys Lys Val Leu Glu
210 215 220
Val Phe Val Glu Lys Gly Met Gln Asn Gly Gln Lys Ile Thr Phe Pro
225 230 235 240
Gly Glu Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Ile Phe
245 250 255
Val Leu Gln Gln Lys Glu His Pro Lys Phe Lys Arg Lys Gly Asp Asp
260 265 270
Leu Phe Tyr Glu His Thr Leu Thr Leu Thr Glu Ser Leu Cys Gly Phe
275 280 285
Gln Phe Val Val Thr His Leu Asp Asn Arg Gln Leu Leu Ile Lys Ser
290 295 300
Asn Pro Gly Glu Val Val Lys Pro Asp Ser Phe Lys Ala Ile Asn Asp
305 310 315 320
Glu Gly Met Pro Met Tyr Gln Arg Pro Phe Met Lys Gly Lys Leu Tyr
325 330 335
Ile His Phe Ser Val Glu Phe Pro Asp Ser Leu Ser Pro Glu Gln Cys
340 345 350
Lys Ala Leu Glu Ala Val Leu Pro Pro Lys Pro Val Ser Gln Tyr Thr
355 360 365
Asp Met Glu Leu Asp Glu Cys Glu Glu Thr Met Pro Tyr Asp Val Asn
370 375 380
Ile Glu Ala Glu Met Arg Arg Arg Gln Gln Gln His Gln Glu Ala Tyr
385 390 395 400
Asp Glu Asp Glu Asp Met Pro Gly Gly Ala Gln Arg Val Gln Cys Ala
405 410 415
Gln Gln
<210>13
<211>1269
<212>DNA
<213>玉米
<400>13
atgtttggac gcatgccaag gaagagtagt aacaatacca agtattacga ggttcttggt 60
gtgtctaaga ccgcaagtca ggatgagctt aagaaagcat acagaaaagc tgccataaaa 120
aaccatcctg ataagggtgg agaccctgag aagtttaaag agctgtctca agcttatgat 180
gttcttagtg acccggagaa gagggagatc tatgaccagt atggagaaga tgcccttaag 240
gaaggaatgg gaggaggcag cagcagtgat ttccatagcc ctttcgacat ttttgagcaa 300
ctttttccgg gttctagcac ctttgggggt ggtagctcaa gaggacgcag acaaaagcgt 360
ggtgaagatg tggtgcatac tatgaaggtt tccttagacg atctgtacaa tgggacaacc 420
aagaaactat ctttatcgcg gagtgctttg tgctccaagt gcaaggggaa aggatccaag 480
agtggggcat caggaacatg ccatggttgt cgtggtgctg gaatgagaac aatcacaaga 540
cagataggcc ttggcatgat ccaacagatg aacactgttt gccctgaatg caaaggatca 600
ggtgagatca taagtgacaa ggacaaatgc ccaagctgta aaggaaacaa ggtagtccag 660
gagaagaagg tgttagaggt tcatgtggag aaaggaatgc aacataacca aaagattgta 720
ttccagggtc aagctgatga agctcctgat acggttacag gagacattgt ttttgtcttg 780
caacttaaag accatccaaa atttaagagg atgtacgatg acttatatgt tgagcacaca 840
atctctctca ccgaagcatt gtgtggcttc cagtttgttc ttactcatct tgatgggcga 900
cagcttctga tcaaatctga ccccggggag gttattaaac caggtcaaca caaggccatt 960
aacgatgaag gtatgcctca gcatggccgt cctttcatga agggccgtct gtttgttgaa 1020
ttcaacgtgg tgtttcccga gcctggtgcg ctctcccctg cccagtgccg atcgttggag 1080
aagatccttc cgccgaaacc agggagccaa ctgtcggaca tggagctgga ccagtgcgag 1140
gagaccaccc ttcacgatgt caacattgaa gaggagatga ggcgcaggca gcagcagaag 1200
aagcaggaag cctacgatga agacgaggag gaggatgctc aaccaagggt gcaatgtgcc 1260
cagcagtaa 1269
<210>14
<211>422
<212>PRT
<213>玉米
<400>14
Met Phe Gly Arg Met Pro Arg Lys Ser Ser Asn Asn Thr Lys Tyr Tyr
1 5 10 15
Glu Val Leu Gly Val Ser Lys Thr Ala Ser Gln Asp Glu Leu Lys Lys
20 25 30
Ala Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp
35 40 45
Pro Glu Lys Phe Lys Glu Leu Ser Gln Ala Tyr Asp Val Leu Ser Asp
50 55 60
Pro Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys
65 70 75 80
Glu Gly Met Gly Gly Gly Ser Ser Ser Asp Phe His Ser Pro Phe Asp
85 90 95
Ile Phe Glu Gln Leu Phe Pro Gly Ser Ser Thr Phe Gly Gly Gly Ser
100 105 110
Ser Arg Gly Arg Arg Gln Lys Arg Gly Glu Asp Val Val His Thr Met
115 120 125
Lys Val Ser Leu Asp Asp Leu Tyr Asn Gly Thr Thr Lys Lys Leu Ser
130 135 140
Leu Ser Arg Ser Ala Leu Cys Ser Lys Cys Lys Gly Lys Gly Ser Lys
145 150 155 160
Ser Gly Ala Ser Gly Thr Cys His Gly Cys Arg Gly Ala Gly Met Arg
165 170 175
Thr Ile Thr Arg Gln Ile Gly Leu Gly Met Ile Gln Gln Met Asn Thr
180 185 190
Val Cys Pro Glu Cys Lys Gly Ser Gly Glu Ile Ile Ser Asp Lys Asp
195 200 205
Lys Cys Pro Ser Cys Lys Gly Asn Lys Val Val Gln Glu Lys Lys Val
210 215 220
Leu Glu Val His Val Glu Lys Gly Met Gln His Asn Gln Lys Ile Val
225 230 235 240
Phe Gln Gly Gln Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile
245 250 255
Val Phe Val Leu Gln Leu Lys Asp His Pro Lys Phe Lys Arg Met Tyr
260 265 270
Asp Asp Leu Tyr Val Glu His Thr Ile Ser Leu Thr Glu Ala Leu Cys
275 280 285
Gly Phe Gln Phe Val Leu Thr His Leu Asp Gly Arg Gln Leu Leu Ile
290 295 300
Lys Ser Asp Pro Gly Glu Val Ile Lys Pro Gly Gln His Lys Ala Ile
305 310 315 320
Asn Asp Glu Gly Met Pro Gln His Gly Arg Pro Phe Met Lys Gly Arg
325 330 335
Leu Phe Val Glu Phe Asn Val Val Phe Pro Glu Pro Gly Ala Leu Ser
340 345 350
Pro Ala Gln Cys Arg Ser Leu Glu Lys Ile Leu Pro Pro Lys Pro Gly
355 360 365
Ser Gln Leu Ser Asp Met Glu Leu Asp Gln Cys Glu Glu Thr Thr Leu
370 375 380
His Asp Val Asn Ile Glu Glu Glu Met Arg Arg Arg Gln Gln Gln Lys
385 390 395 400
Lys Gln Glu Ala Tyr Asp Glu Asp Glu Glu Glu Asp Ala Gln Pro Arg
405 410 415
Val Gln Cys Ala Gln Gln
420
<210>15
<211>1266
<212>DNA
<213>小麦(Triticum aestivum)
<400>15
atgttcgggc gcgggccgcc gaagaagagc gacagcacgc gctactacga gatcctgggc 60
gtgcccaagg acgcgtccca ggacgacctc aagaaggcct accgcaaggc cgccatcaag 120
aaccaccccg acaagggagg cgacccagag aagttcaagg agctagctca ggcttatgag 180
gttctgagtg atcctgagaa gcgagagatc tatgaccagt atggtgagga tgccctcaag 240
gagggaatgg gaggtggagg aatgcatgat ccttttgaca tcttccagtc attctttggt 300
ggtggcggca accccttcgg aggtggcggg agcagtaggg gcaggcggca gcgcaggggt 360
gaggatgtgg ttcatcctct gaaggttagc cttgaggaac tgtacaacgg aacatcaaag 420
aagctctctc ttgcccgcaa tgtgctctgc tcgaagtgca atggcaaggg gtcaaagtcc 480
ggggcttcga tgaagtgtgc cggctgccaa ggtgctggtt acaaggtgca gataaggcag 540
ctgggaccag gaatgattca gcaaatgcag cagccttgca atgagtgcag gggaagtggg 600
gagaccatca gcgacaagga tcgctgtggg cagtgcaaag gcgagaaggt ggtgcacgag 660
aagaaagtcc tggaggtggt ggtcgagaag ggaatgcagc atgggcagaa gatcaccttc 720
cccggcgagg cggatgaagc gcctgatact gttactggag acataatctt cgtcctccag 780
cagaaggagc accccaaatt caagcggaag ggcgatgacc tcttctacga gcacaccctg 840
accctgaccg aggcactgtg tggcttccag tatgtcctgg ctcatttgga cggcaggcag 900
ctgctcatca agtccaaccc tggcgaagtc gtcaagcctg attcgttcaa ggcgatcaac 960
gacgagggca tgcccatgta ccagaggccg ttcatgaagg gcaagctgta catccacttc 1020
acggttgatt ttcccgactc gctgagcctg gaccagtgca aggcgctcga gactgtcctg 1080
ccgcccaagc cggcgtcgca gtacacggac atggagctgg acgagtgcga ggagacgatg 1140
gcctacgaca ttgacatcga ggaggagatg cggaggcgac agcagcagca ggcacaggag 1200
gcctacgacg aggacgagga catgcccggt ggcggcggcc agcgggtgca gtgcgcccag 1260
cagtag 1266
<210>16
<211>421
<212>PRT
<213>小麦
<400>16
Met Phe Gly Arg Gly Pro Pro Lys Lys Ser Asp Ser Thr Arg Tyr Tyr
1 5 10 15
Glu Ile Leu Gly Val Pro Lys Asp Ala Ser Gln Asp Asp Leu Lys Lys
20 25 30
Ala Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp
35 40 45
Pro Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp
50 55 60
Pro Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys
65 70 75 80
Glu Gly Met Gly Gly Gly Gly Met His Asp Pro Phe Asp Ile Phe Gln
85 90 95
Ser Phe Phe Gly Gly Gly Gly Asn Pro Phe Gly Gly Gly Gly Ser Ser
100 105 110
Arg Gly Arg Arg Gln Arg Arg Gly Glu Asp Val Val His Pro Leu Lys
115 120 125
Val Ser Leu Glu Glu Leu Tyr Asn Gly Thr Ser Lys Lys Leu Ser Leu
130 135 140
Ala Arg Asn Val Leu Cys Ser Lys Cys Asn Gly Lys Gly Ser Lys Ser
145 150 155 160
Gly Ala Ser Met Lys Cys Ala Gly Cys Gln Gly Ala Gly Tyr Lys Val
165 170 175
Gln Ile Arg Gln Leu Gly Pro Gly Met Ile Gln Gln Met Gln Gln Pro
180 185 190
Cys Asn Glu Cys Arg Gly Ser Gly Glu Thr Ile Ser Asp Lys Asp Arg
195 200 205
Cys Gly Gln Cys Lys Gly Glu Lys Val Val His Glu Lys Lys Val Leu
210 215 220
Glu Val Val Val Glu Lys Gly Met Gln His Gly Gln Lys Ile Thr Phe
225 230 235 240
Pro Gly Glu Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Ile
245 250 255
Phe Val Leu Gln Gln Lys Glu His Pro Lys Phe Lys Arg Lys Gly Asp
260 265 270
Asp Leu Phe Tyr Glu His Thr Leu Thr Leu Thr Glu Ala Leu Cys Gly
275 280 285
Phe Gln Tyr Val Leu Ala His Leu Asp Gly Arg Gln Leu Leu Ile Lys
290 295 300
Ser Asn Pro Gly Glu Val Val Lys Pro Asp Ser Phe Lys Ala Ile Asn
305 310 315 320
Asp Glu Gly Met Pro Met Tyr Gln Arg Pro Phe Met Lys Gly Lys Leu
325 330 335
Tyr Ile His Phe Thr Val Asp Phe Pro Asp Ser Leu Ser Leu Asp Gln
340 345 350
Cys Lys Ala Leu Glu Thr Val Leu Pro Pro Lys Pro Ala Ser Gln Tyr
355 350 365
Thr Asp Met Glu Leu Asp Glu Cys Glu Glu Thr Met Ala Tyr Asp Ile
370 375 380
Asp Ile Glu Glu Glu Met Arg Arg Arg Gln Gln Gln Gln Ala Gln Glu
385 390 395 400
Ala Tyr Asp Glu Asp Glu Asp Met Pro Gly Gly Gly Gly Gln Arg Val
405 410 415
Gln Cys Ala Gln Gln
420
<210>17
<211>1260
<212>DNA
<213>拟南芥(Arabidopsis thaliana)
<400>17
atgtttggaa gaggaccttc aaggaagagc gataacacaa agttctacga gatccttggt 60
gttcctaaga ccgcagcacc agaagatctc aagaaagctt ataagaaagc cgctatcaaa 120
aaccatcctg ataagggtgg tgatcccgaa aagtttaaag agttagcaca ggcttatgaa 180
gttttaagtg atcctgagaa gcgtgagatc tatgatcaat atggggaaga tgcactcaag 240
gaaggaatgg gtggtggagg tggtggacac gatccatttg atatcttctc ttccttcttt 300
ggtagtggtg gacacccatt cggaagtcat agccggggaa ggaggcagag gcgtggtgaa 360
gatgttgttc atcccttgaa ggtttcctta gaggatgttt atctcggaac aacaaagaag 420
ctctcacttt ctaggaaggc tttgtgctca aagtgtaacg gcaagggttc aaagtctgga 480
gcttcactga aatgtggtgg ctgtcaaggc tcgggaatga agatctcgat caggcagttt 540
ggacctggaa tgatgcagca ggtgcagcat gcttgtaatg attccaaagg cacaggagag 600
accatcaatg atcgggacag gtgtccacaa tgcaaaggag agaaggttgt ctctgagaag 660
aaggtgcttg aagtaaatgt ggagaaggga atgcaacaca atcagaagat cacattcagt 720
ggacaagccg atgaagcgcc tgatactgtc accggagata tagtgtttgt cattcagcag 780
aaggagcacc caaagttcaa aagaaagggt gaggatctct ttgtggagca caccatctct 840
ctaaccgagg ccttgtgtgg cttccagttt gtcttgaccc atttggacaa aagacagctt 900
ctcatcaaat ccaagcccgg agaggtcgtc aaacctgatt catacaaggc gataagtgat 960
gagggaatgc caatatacca aagtccgttc atgaagggta agctatacat tcacttcacg 1020
gttgaattcc cggaatcgct gagcccggat cagacaaagg ccattgaagc agttttgcca 1080
aagccaacca aggcagctat aagcgatatg gaaatagacg actgcgaaga gacgactctg 1140
caagatgtga acattgagga tgagatgaaa aggaaggcgc aagctcaaag agaggcttat 1200
gatgtcgatg aggaagatca cccaggcggt gctcaccgtg tgcaatgtgc ccagcagtga 1260
<210>18
<211>419
<212>PRT
<213>拟南芥
<400>18
Met Phe Gly Arg Gly Pro Ser Arg Lys Ser Asp Asn Thr Lys Phe Tyr
1 5 10 15
Glu Ile Leu Gly Val Pro Lys Thr Ala Ala Pro Glu Asp Leu Lys Lys
20 25 30
Ala Tyr Lys Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp
35 40 45
Pro Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp
50 55 60
Pro Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys
65 70 75 80
Glu Gly Met Gly Gly Gly Gly Gly Gly His Asp Pro Phe Asp Ile Phe
85 90 95
Ser Ser Phe Phe Gly Ser Gly Gly His Pro Phe Gly Ser His Ser Arg
100 105 110
Gly Arg Arg Gln Arg Arg Gly Glu Asp Val Val His Pro Leu Lys Val
115 120 125
Ser Leu Glu Asp Val Tyr Leu Gly Thr Thr Lys Lys Leu Ser Leu Ser
130 135 140
Arg Lys Ala Leu Cys Ser Lys Cys Asn Gly Lys Gly Ser Lys Ser Gly
145 150 155 160
Ala Ser Met Lys Cys Gly Gly Cys Gln Gly Ser Gly Met Lys Ile Ser
165 170 175
Ile Arg Gln Phe Gly Pro Gly Met Met Gln Gln Val Gln His Ala Cys
180 185 190
Asn Asp Cys Lys Gly Thr Gly Glu Thr Ile Asn Asp Arg Asp Arg Cys
195 200 205
Pro Gln Cys Lys Gly Glu Lys Val Val Ser Glu Lys Lys Val Leu Glu
210 215 220
Val Asn Val Glu Lys Gly Met Gln His Asn Gln Lys Ile Thr Phe Ser
225 230 235 240
Gly Gln Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Val Phe
245 250 255
Val Ile Gln Gln Lys Glu His Pro Lys Phe Lys Arg Lys Gly Glu Asp
260 265 270
Leu Phe Val Glu His Thr Ile Ser Leu Thr Glu Ala Leu Cys Gly Phe
275 280 285
Gln Phe Val Leu Thr His Leu Asp Lys Arg Gln Leu Leu Ile Lys Ser
290 295 300
Lys Pro Gly Glu Val Val Lys Pro Asp Ser Tyr Lys Ala Ile Ser Asp
305 310 315 320
Glu Gly Met Pro Ile Tyr Gln Arg Pro Phe Met Lys Gly Lys Leu Tyr
325 330 335
Ile His Phe Thr Val Glu Phe Pro Glu Ser Leu Ser Pro Asp Gln Thr
340 345 350
Lys Ala Ile Glu Ala Val Leu Pro Lys Pro Thr Lys Ala Ala Ile Ser
355 360 365
Asp Met Glu Ile Asp Asp Cys Glu Glu Thr Thr Leu His Asp Val Asn
370 375 380
Ile Glu Asp Glu Met Lys Arg Lys Ala Gln Ala Gln Arg Glu Ala Tyr
385 390 395 400
Asp Asp Asp Glu Glu Asp His Pro Gly Gly Ala Gln Arg Val Gln Cys
405 410 415
Ala Gln Gln
<210>19
<211>1263
<212>DNA
<213>拟南芥
<400>19
atgttcggta gaggaccctc gaagaagagc gacaacacta agttctacga gatcttaggt 60
gttcctaaga gcgcttcacc agaagatctc aagaaagctt acaaaaaagc cgctatcaag 120
aatcatcctg ataagggtgg agatcccgag aaggtgaata atttcttaga tccgtatgaa 180
gtgcttagtg acccggagaa gcgtgagatt tatgaccagt atggagagga tgcactcaag 240
gaaggaatgg gtggtggagg aggtggacat gatccatttg atattttctc atccttcttt 300
ggtggaggcc cctttggagg tgagtctcct tggacactgt ggcagaggcg tggtgaggat 360
gttgttcatc ccttgaaggt atctcttgag gatgtgtacc ttggtacaat gaagaagctt 420
tcactttcta ggaatgctct ctgctctaag tgtaacgggt tagtacattc gactcgatcc 480
tccttgaaat gtggagggtg tcagggatct ggtatgaagg tgtctattag gcagcttgga 540
cctggaatga tccagcagat gcagcatgca tgtaatgaat gcaaagggac aggtgagacc 600
atcaatgatc gggacaggtg tccacaatgc aaaggagaca aggtcattcc tgagaagaag 660
gtgcttgaag tgaatgtgga gaagggaatg caacacagtc agaagatcac atttgaagga 720
caagcagatg aagcggtatc tactctcata catttaatag tgtttgtcct tcagcagaaa 780
gagcacccaa agttcaagag aaagggagaa gacctctttg tggagcacac actttctcta 840
accgaagctt tgtgtggctt ccaatttgtt ctgactcact tggatggcag aagtcttctc 900
attaaatcta atcctgggga ggtcgtgaaa cctggtacgt attcagatgc atcgtatgaa 960
ggaatgccga tataccagag gccattcatg aagggtaagc tctacatcca cttcacagtg 1020
gagttcccgg actcgttgag cccagatcag accaaagcac tggaagctgt tctacctaag 1080
ccgtcaacag ctcagttgag tgacatggag atagatgaat gcgaggagac cacgctccac 1140
gatgtcaaca ttgaggatga gatgaggagg aaggcacaag ctcaaagaga ggcttatgat 1200
gatgacgatg aagatgatga ccatccgggt ggtgctcaaa gggtgcaatg tgcccagcag 1260
taa 1263
<210>20
<211>420
<212>PRT
<213>拟南芥
<400>20
Met Phe Gly Arg Gly Pro Ser Lys Lys Ser Asp Asn Thr Lys Phe Tyr
1 5 10 15
Glu Ile Leu Gly Val Pro Lys Ser Ala Ser Pro Glu Asp Leu Lys Lys
20 25 30
Ala Tyr Lys Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp
35 40 45
Pro Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp
50 55 60
Pro Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys
65 70 75 80
Glu Gly Met Gly Gly Gly Gly Gly Gly His Asp Pro Phe Asp Ile Phe
85 90 95
Ser Ser Phe Phe Gly Gly Gly Pro Phe Gly Gly Asn Thr Ser Arg Gln
100 105 110
Arg Arg Gln Arg Arg Gly Glu Asp Val Val His Pro Leu Lys Val Ser
115 120 125
Leu Glu Asp Val Tyr Leu Gly Thr Met Lys Lys Leu Ser Leu Ser Arg
130 135 140
Asn Ala Leu Cys Ser Lys Cys Asn Gly Lys Gly Ser Lys Ser Gly Ala
145 150 155 160
Ser Leu Lys Cys Gly Gly Cys Gln Gly Ser Gly Met Lys Val Ser Ile
165 170 175
Arg Gln Leu Gly Pro Gly Met Ile Gln Gln Met Gln His Ala Cys Asn
180 185 190
Glu Cys Lys Gly Thr Gly Glu Thr Ile Asn Asp Arg Asp Arg Cys Pro
195 200 205
Gln Cys Lys Gly Asp Lys Val Ile Pro Glu Lys Lys Val Leu Glu Val
210 215 220
Asn Val Glu Lys Gly Met Gln His Ser Gln Lys Ile Thr Phe Glu Gly
225 230 235 240
Gln Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Val Phe Val
245 250 255
Leu Gln Gln Lys Glu His Pro Lys Phe Lys Arg Lys Gly Glu Asp Leu
260 265 270
Phe Val Glu His Thr Leu Ser Leu Thr Glu Ala Leu Cys Gly Phe Gln
275 280 285
Phe Val Leu Thr His Leu Asp Gly Arg Ser Leu Leu Ile Lys Ser Asn
290 295 300
Pro Gly Glu Val Val Lys Pro Asp Set Tyr Lys Ala Ile Ser Asp Glu
305 310 315 320
Gly Met Pro Ile Tyr Gln Arg Pro Phe Met Lys Gly Lys Leu Tyr Ile
325 330 335
His Phe Thr Val Glu Phe Pro Asp Ser Leu Ser Pro Asp Gln Thr Lys
340 345 350
Ala Leu Glu Ala Val Leu Pro Lys Pro Ser Thr Ala Gln Leu Ser Asp
355 360 365
Met Glu Ile Asp Glu Cys Glu Glu Thr Thr Leu His Asp Val Asn Ile
370 375 380
Glu Asp Glu Met Arg Arg Lys Ala Gln Ala Gln Arg Glu Ala Tyr Asp
385 390 395 400
Asp Asp Asp Glu Asp Asp Asp His Pro Gly Gly Ala Gln Arg Val Gln
405 410 415
Cys Ala Gln Gln
420
<210>21
<211>1254
<212>DNA
<213>大洋洲滨藜(Atriplex nummularia)
<400>21
atgtttggaa gagcaccaaa gaagagtgat agcaccagat attacgagat cttaggcgta 60
ccaaaagatg catctcctga agatttgaag aaggcttata aaaaagctgc cattaaaaat 120
catcctgaca agggaggtga tcccgagaag tttaaagagc tagctcatgc ttatgaggtc 180
ctcagtgatc ccgaaaagcg tgagatctat gatcaatatg gtgaggatgc acttaaggaa 240
gaatgggtg gaggtggcgg tatgcatgat ccattcgaca tcttccaatc cttctttgga 300
ggaagtccat ttggtggtgt tggttctagc cgaggaagaa ggcaaaggcg gggagaagat 360
gtagttcatc ctcttaaggt ttcactcgag gatctcttta ccggtacaac aaagaagctc 420
tcactctctc gcaatgtaat ttgttcaaag tgtactggca aaggatcaaa atcgggagct 480
tctatgaagt gttctggatg tcaaggtact ggtatgaagg tttctatcag acatctggga 540
ccctcaatga tccagcagat gcagcaccct tgtaatgaat gcaaaggaac tggagagacg 600
attaatgaca aagatcgttg ccctcagtgc aaaggtgaga aggttgtgca ggagaagaag 660
gtcttagagg ttgttgtgga gaagggcatg caacatggac agaaaattac tttccctgga 720
gaggctgatg aagctcctga tactgtcact ggagatatag tctttgtcct gcagcagaaa 780
gagcacccta agttcaagag aaagggtgaa gatctcttct acgagcacac tctaagcctg 840
actgaagctc tttgcggctt tagatttgtg ctgactcacc ttgatggaag gcaacttctt 900
atcaaatcaa acctgggaga agttgtcaag cctgatcaat tcaaggcaat tgaggatgag 960
ggtatgccta tataccaaag gccgttcatg aagggcaaga tgtacatcca tttcacagtg 1020
gagttccccg attcgttaaa ccctgatcaa gttaaatcct tggaagcgat ccttcctcct 1080
aagccatcaa tgtctctcac atacatggag ttagatgaat gtgaagagac aacactgcat 1140
aatgtcaaca ttgaagaaga gatgaaaagg aagcagacac aagcacagca ggaggcatac 1200
gatgaagatg acgaacctgc cggtggtcag agggtccaat gtgctcaaca gtga 1254
<210>22
<211>417
<212>PRT
<213>大洋洲滨藜
<400>22
Met Phe Gly Arg Ala Pro Lys Lys Ser Asp Ser Thr Arg Tyr Tyr Glu
1 5 10 15
Ile Leu Gly Val Pro Lys Asp Ala Ser Pro Glu Asp Leu Lys Lys Ala
20 25 30
Tyr Lys Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala His Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Gly Gly Gly Gly Met His Asp Pro Phe Asp Ile Phe Gln
85 90 95
Ser Phe Phe Gly Gly Ser Pro Phe Gly Gly Val Gly Ser Ser Arg Gly
100 105 110
Arg Arg Gln Arg Arg Gly Glu Asp Val Val His Pro Leu Lys Val Ser
115 120 125
Leu Glu Asp Leu Phe Thr Gly Thr Thr Lys Lys Leu Ser Leu Ser Arg
130 135 140
Asn Val Ile Cys Ser Lys Cys Thr Gly Lys Gly Ser Lys Ser Gly Ala
145 150 155 160
Ser Met Lys Cys Ser Gly Cys Gln Gly Thr Gly Met Lys Val Ser Ile
165 170 175
Arg His Leu Gly Pro Ser Met Ile Gln Gln Met Gln His Pro Cys Asn
180 185 190
Glu Cys Lys Gly Thr Gly Glu Thr Ile Asn Asp Lys Asp Arg Cys Pro
195 200 205
Gln Cys Lys Gly Glu Lys Val Val Gln Glu Lys Lys Val Leu Glu Val
210 215 220
Val Val Glu Lys Gly Met Gln His Gly Gln Lys Ile Thr Phe Pro Gly
225 230 235 240
Glu Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Val Phe Val
245 250 255
Leu Gln Gln Lys Glu His Pro Lys Phe Lys Arg Lys Gly Glu Asp Leu
260 265 270
Phe Tyr Glu His Thr Leu Ser Leu Thr Glu Ala Leu Cys Gly Phe Arg
275 280 285
Phe Val Leu Thr His Leu Asp Gly Arg Gln Leu Leu Ile Lys Ser Asn
290 295 300
Leu Gly Glu Val Val Lys Pro Asp Gln Phe Lys Ala Ile Glu Asp Glu
305 310 315 320
Gly Met Pro Ile Tyr Gln Arg Pro Phe Met Lys Gly Lys Met Tyr Ile
325 330 335
His Phe Thr Val Glu Phe Pro Asp Ser Leu Asn Pro Asp Gln Val Lys
340 345 350
Ser Leu Glu Ala Ile Leu Pro Pro Lys Pro Ser Met Ser Leu Thr Tyr
355 360 365
Met Glu Leu Asp Glu Cys Glu Glu Thr Thr Leu His Asn Val Asn Ile
370 375 380
Glu Glu Glu Met Lys Arg Lys Gln Thr Gln Ala Gln Gln Glu Ala Tyr
385 390 395 400
Asp Glu Asp Asp Glu Pro Ala Gly Gly Gln Arg Val Gln Cys Ala Gln
405 410 415
Gln
<210>23
<211>1242
<212>DNA
<213>黄瓜(Cucumis sativus)
<400>23
atgtttggaa ggccgaagaa gagcgataat accaaatatt atgagattct tggagtctcg 60
aagaatgcgt cgcaagacga tctaaagaag gcttatagaa aggccgccat caagaaccat 120
cctgataaag gtggcgaccc tgaaaaattc aaggagttag cacaagccta cgaggtgctg 180
agtgatccag agaaacgtga gatatatgat caatatggcg aggatgccct caaggaagga 240
atgggaggtg gcggtggtca tgatccattt gacatattcc agtctttctt tggtggaagc 300
ccgtttggtg gtggtggaag cagcagaggc cgaaggcaga gaaggggaga ggatgttatc 360
catcctctca aggtctcgtt ggaagatctc tacaacggta cttcaaagaa gctctctctt 420
tcacgtaatg taatttgctc aaagtgcaag ggtaagggtt ctaaatctgg tgcttcaatg 480
aagtgtcctg gctgtcaagg ttctggtatg aaagtttcca tcagacacct tggcccctct 540
atgattcagc aaatgcagca tccttgcaat gaatgtaaag gaactggtga gaccatcaat 600
gataaagatc gctgctcaca atgcaagggt gaaaaggttg ttcaggagaa aaaagttttg 660
gaagttattg tggagaaggg tatgcaaaat gcacaaaaga ttacattccc tggtgaagca 720
gatgaagcgc ccgacactgt tactggggac attgtctttg tcctacaaca aaaagagcac 780
cccaagttta agagaaaggg cgatgacctc tttgtagagc ataccttgtc tctcgtcgag 840
tctctgtgtg gtttccaatt tattctgact catttggatg gccgacagct actcatcaaa 900
tcacttcccg gtgaagtagt gaagcctgac caattcaagg ccataaacga tgagggtatg 960
cctatgtacc agaggccatt catgaagggc aaactttaca tccacttcag tgttgagttc 1020
ccagactcct tgaaccccga acagtgcaag gcgctggagg gcgttctgcc tcccaggacc 1080
tcagtgcagc tctcagatat ggaattggat gaatgtgaag agaccactct ccacgatgtc 1140
aacattgaag aggagatgcg caggaagcaa gcacaagagg catacgatga agatgaggat 1200
atgcacggtg gtgcacagag agtgcagtgt gctcaacaat ga 1242
<210>24
<211>413
<212>PRT
<213>黄瓜
<400>24
Met Phe Gly Arg Pro Lys Lys Ser Asp Asn Thr Lys Tyr Tyr Glu Ile
1 5 10 15
Leu Gly Val Ser Lys Asn Ala Ser Gln Asp Asp Leu Lys Lys Ala Tyr
20 25 30
Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro Glu
35 40 45
Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Pro Glu
50 55 60
Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu Gly
65 70 75 80
Met Gly Gly Gly Gly Gly His Asp Pro Phe Asp Ile Phe Gln Ser Phe
85 90 95
Phe Gly Gly Ser Pro Phe Gly Gly Gly Gly Ser Ser Arg Gly Arg Arg
100 105 110
Gln Arg Arg Gly Glu Asp Val Ile His Pro Leu Lys Val Ser Leu Glu
115 120 125
Asp Leu Tyr Asn Gly Thr Ser Lys Lys Leu Ser Leu Ser Arg Asn Val
130 135 140
Ile Cys Ser Lys Cys Lys Gly Lys Gly Ser Lys Ser Gly Ala Ser Met
145 150 155 160
Lys Cys Pro Gly Cys Gln Gly Ser Gly Met Lys Val Ser Ile Arg His
165 170 175
Leu Gly Pro Ser Met Ile Gln Gln Met Gln His Pro Cys Asn Glu Cys
180 185 190
Lys Gly Thr Gly Glu Thr Ile Asn Asp Lys Asp Arg Cys Ser Gln Cys
195 200 205
Lys Gly Glu Lys Val Val Gln Glu Lys Lys Val Leu Glu Val Ile Val
210 215 220
Glu Lys Gly Met Gln Asn Ala Gln Lys Ile Thr Phe Pro Gly Glu Ala
225 230 235 240
Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Val Phe Val Leu Gln
245 250 255
Gln Lys Glu His Pro Lys Phe Lys Arg Lys Gly Asp Asp Leu Phe Val
260 265 270
Glu His Thr Leu Ser Leu Val Glu Ser Leu Cys Gly Phe Gln Phe Ile
275 280 285
Leu Thr His Leu Asp Gly Arg Gln Leu Leu Ile Lys Ser Leu Pro Gly
290 295 300
Glu Val Val Lys Pro Asp Gln Phe Lys Ala Ile Asn Asp Glu Gly Met
305 310 315 320
Pro Met Tyr Gln Arg Pro Phe Met Lys Gly Lys Leu Tyr Ile His Phe
325 330 335
Ser Val Glu Phe Pro Asp Ser Leu Asn Pro Glu Gln Cys Lys Ala Leu
340 345 350
Glu Gly Val Leu Pro Pro Arg Thr Ser Val Gln Leu Ser Asp Met Glu
355 360 365
Leu Asp Glu Cys Glu Glu Thr Thr Leu His Asp Val Asn Ile Glu Glu
370 375 380
Glu Met Arg Arg Lys Gln Ala Gln Glu Ala Tyr Asp Glu Asp Glu Asp
385 390 395 400
Met His Gly Gly Ala Gln Arg Val Gln Cys Ala Gln Gln
405 410
<210>25
<211>1257
<212>DNA
<213>胡萝卜(Daucus carota)
<400>25
atgtttggga gagcaccaaa gaagagtgac aatacaaagt actatgaaat tcttggtgtc 60
ccaaaaacag catcacctga tgatctgaag aaagcttaca ggaaggctgc tatcaagaat 120
catcctgata agggtggcga tcctgaaaag tttaaagagc ttgcgcaagc atatgaggtt 180
ctgagtgacc cagagaagcg tgaaatctat gatcagtatg gagaggatgc tctcaaggag 240
ggaatgggtg gtggtggagg tggtggccat gacccatttg acattttcca atccttcttt 300
ggtggcagcc cgtttggtgg aggtggcagc agcagaggac gaaggcaaag aaggggggag 360
gatgtcattc atccccttaa ggtttcactg gaagatcttt gcaatgggac ttccaagaag 420
ctttcccttt cacgtaatgt aatttgttct aaatgcaagg gaaaggggtc caagtcgggt 480
gcttcaatga catgtcctgg ctgccagggt tctggaatga aggtttctat caggcacttg 540
ggcccatcta tgatccagca gatgcagcat ccctgcaatg actgcaaggg tactggagaa 600
acaatcaacg acaaggatcg ctgccctcaa tgcaaaggtc aaaaggttgt gcaggagaag 660
aaagcaatag aagttattgt ggagaagggt atgcaaaacg gacagaagat tacattccct 720
ggagaagctg atgaagcgcc tgacacggtt actggggaca tagtgtttgt gttgcaacaa 780
aaggagcacc ccaagtttaa gaggaagggt gatgatcttt ttgttgaaca ttcattaact 840
ctcagtgaag cactttgtgg cttccaattt actttgactc acctggacgg caggcagctt 900
cttattaaat cccagccagg agaagttatc aagccagatc aatttaaggg gataaatgat 960
gaaggaatgc caatgtatca gaggccattt atgcgaggaa agctttacat tcactttagt 1020
gtagatttcc cagagtcctt gacccctgag cagtgcaaag ctcttgaagc tgtgttacct 1080
ccgaggcctt caattcagat gacagacatg gaactggatg aatgtgaaga aacaacactg 1140
catgatgtga atattgaaga ggagatgcgt cggaaacagc aagctgccca agaggcatat 1200
gacgaagacg aagatatgca tggcggtgct cagagggtgc agtgtgctca acaatga 1257
<210>26
<211>418
<212>PRT
<213>胡萝卜
<400>26
Met Phe Gly Arg Ala Pro Lys Lys Ser Asp Asn Thr Lys Tyr Tyr Glu
1 5 10 15
Ile Leu Gly Val Pro Lys Thr Ala Ser Pro Asp Asp Leu Lys Lys Ala
20 25 30
Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Gly Gly Gly Gly Gly Gly His Asp Pro Phe Asp Ile Phe
85 90 95
Gln Ser Phe Phe Gly Gly Ser Pro Phe Gly Gly Gly Gly Ser Ser Arg
100 105 110
Gly Arg Arg Gln Arg Arg Gly Glu Asp Val Ile His Pro Leu Lys Val
115 120 125
Ser Leu Glu Asp Leu Cys Asn Gly Thr Ser Lys Lys Leu Ser Leu Ser
130 135 140
Arg Asn Val Ile Cys Ser Lys Cys Lys Gly Lys Gly Ser Lys Ser Gly
145 150 155 160
Ala Ser Met Thr Cys Pro Gly Cys Gln Gly Ser Gly Met Lys Val Ser
165 170 175
Ile Arg His Leu Gly Pro Ser Met Ile Gln Gln Met Gln His Pro Cys
180 185 190
Asn Asp Cys Lys Gly Thr Gly Glu Thr Ile Asn Asp Lys Asp Arg Cys
195 200 205
Pro Gln Cys Lys Gly Gln Lys Val Val Gln Glu Lys Lys Ala Ile Glu
210 215 220
Val Ile Val Glu Lys Gly Met Gln Asn Gly Gln Lys Ile Thr Phe Pro
225 230 235 240
Gly Glu Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Val Phe
245 250 255
Val Leu Gln Gln Lys Glu His Pro Lys Phe Lys Arg Lys Gly Asp Asp
260 265 270
Leu Phe Val Glu His Ser Leu Thr Leu Ser Glu Ala Leu Cys Gly Phe
275 280 285
Gln Phe Thr Leu Thr His Leu Asp Gly Arg Gln Leu Leu Ile Lys Ser
290 295 300
Gln Pro Gly Glu Val Ile Lys Pro Asp Gln Phe Lys Gly Ile Asn Asp
305 310 315 320
Glu Gly Met Pro Met Tyr Gln Arg Pro Phe Met Arg Gly Lys Leu Tyr
325 330 335
Ile His Phe Ser Val Asp Phe Pro Glu Ser Leu Thr Pro Glu Gln Cys
340 345 350
Lys Ala Leu Glu Ala Val Leu Pro Pro Arg Pro Ser Ile Gln Met Thr
355 360 365
Asp Met Glu Leu Asp Glu Cys Glu Glu Thr Thr Leu His Asp Val Asn
370 375 380
Ile Glu Glu Glu Met Arg Arg Lys Gln Gln Ala Ala Gln Glu Ala Tyr
385 390 395 400
Asp Glu Asp Glu Asp Met His Gly Gly Ala Gln Arg Val Gln Cys Ala
405 410 415
Gln Gln
<210>27
<211>1254
<212>DNA
<213>大豆(Glycine max)
<400>27
atgtttggga gggcaccgaa gaagagcgat aatacgaggt actacgaaat cctcggcgtc 60
tccaagaacg cttcgcagga tgatctgaag aaggcttaca agaaagccgc cattaagaat 120
caccccgaca agggcggtga tcccgagaag tttaaagagc tggcgcaagc ttatgaggtt 180
ctgagtgacc ctgagaagcg tgagatatat gatcagtatg gtgaagatgc gcttaaggaa 240
ggaatgggtg gtggcggtgg ccatgatcca tttgatatct tttcatcttt ctttggcggt 300
gggagtccct ttggatcagg tggaagtagt cgaggtagga ggcagaggcg cggagaagac 360
gtggttcacc ctctcaaggt ctctttggag gacctttatc ttggaacttc caagaagctc 420
tccctctcca gaaatgttat atgctccaag tgcagtggca agggttctaa gtctggtgct 480
tcgatgaagt gtgctggttg tcaaggaact ggtatgaagg tttctataag acatcttggc 540
ccatccatga ttcagcagat gcagcatgcc tgcaatgaat gtaagggtac tggagaaact 600
atcaatgaca gagatcgctg cccacagtgc aagggagaga aggttgtgca ggagaagaaa 660
gtccttgaag ttattgtaga aaaggggatg cagaatgggc agaagataac attccctggc 720
gaagctgatg aagcgccgga cacaattact ggggatatcg tctttgtcct tcagcagaag 780
gaacatccca aattcaaaag aaaggctgaa gatctttttg tagagcacac tttgtccctt 840
accgaggcct tgtgtggctt ccaatttgtg ctgactcact tggatagccg tcagcttctt 900
attaaatcaa atcccgggga agttgtgaag cctgattcat acaaggctat aaatgatgag 960
ggaatgccca tgtatcagag gccattcatg aaggggaaac tttacattca cttcactgtg 1020
gagtttccag attctctaaa ccctgatcaa gttaaggcct tggaggctgt tctgccacca 1080
aagccttctt cacaattgac agacatggag ctggatgaat gtgaggaaac tacactccat 1140
gatgtcaaca tggaggagga gactaggagg aagcagcaac aagctcagga ggcatatgat 1200
gaggatgatg acatgcctgg tggtgcacag agggtacagt gcgcccagca gtaa 1254
<210>28
<211>417
<212>PRT
<213>大豆
<400>28
Met Phe Gly Arg Ala Pro Lys Lys Ser Asp Asn Thr Arg Tyr Tyr Glu
1 5 10 15
Ile Leu Gly Val Ser Lys Asn Ala Ser Gln Asp Asp Leu Lys Lys Ala
20 25 30
Tyr Lys Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Gly Gly Gly Gly His Asp Pro Phe Asp Ile Phe Ser Ser
85 90 95
Phe Phe Gly Gly Gly Ser Pro Phe Gly Ser Gly Gly Ser Ser Arg Gly
100 105 110
Arg Arg Gln Arg Arg Gly Glu Asp Val Val His Pro Leu Lys Val Ser
115 120 125
Leu Glu Asp Leu Tyr Leu Gly Thr Ser Lys Lys Leu Ser Leu Ser Arg
130 135 140
Asn Val Ile Cys Ser Lys Cys Ser Gly Lys Gly Ser Lys Ser Gly Ala
145 150 155 160
Ser Met Lys Cys Ala Gly Cys Gln Gly Thr Gly Met Lys Val Ser Ile
165 170 175
Arg His Leu Gly Pro Ser Met Ile Gln Gln Met Gln His Ala Cys Asn
180 185 190
Glu Cys Lys Gly Thr Gly Glu Thr Ile Asn Asp Arg Asp Arg Cys Pro
195 200 205
Gln Cys Lys Gly Glu Lys Val Val Gln Glu Lys Lys Val Leu Glu Val
210 215 220
Ile Val Glu Lys Gly Met Gln Asn Gly Gln Lys Ile Thr Phe Pro Gly
225 230 235 240
Glu Ala Asp Glu Ala Pro Asp Thr Ile Thr Gly Asp Ile Val Phe Val
245 250 255
Leu Gln Gln Lys Glu His Pro Lys Phe Lys Arg Lys Ala Glu Asp Leu
260 265 270
Phe Val Glu His Thr Leu Ser Leu Thr Glu Ala Leu Cys Gly Phe Gln
275 280 285
Phe Val Leu Thr His Leu Asp Ser Arg Gln Leu Leu Ile Lys Ser Asn
290 295 300
Pro Gly Glu Val Val Lys Pro Asp Ser Tyr Lys Ala Ile Asn Asp Glu
305 310 315 320
Gly Met Pro Met Tyr Gln Arg Pro Phe Met Lys Gly Lys Leu Tyr Ile
325 330 335
His Phe Thr Val Glu Phe Pro Asp Ser Leu Asn Pro Asp Gln Val Lys
340 345 350
Ala Leu Glu Ala Val Leu Pro Pro Lys Pro Ser Ser Gln Leu Thr Asp
355 360 365
Met Glu Leu Asp Glu Cys Glu Glu Thr Thr Leu His Asp Val Asn Met
370 375 380
Glu Glu Glu Thr Arg Arg Lys Gln Gln Gln Ala Gln Glu Ala Tyr Asp
385 390 395 400
Glu Asp Asp Asp Met Pro Gly Gly Ala Gln Arg Val Gln Cys Ala Gln
405 410 415
Gln
<210>29
<211>1248
<212>DNA
<213>橡胶(Hevea brasiliensis)
<400>29
atgtttggaa gagcacccaa aaaaagcgat aacaccaagt actatgagat tcttggggtc 60
tcaaagaacg cttcacagga tgatctaaag aaggcttata gaaaagctgc catcaagaac 120
catcctgaca agggtggtga tcctgaaaag tttaaagagt tggcccaagc ttatgaggtt 180
ttgagtgatc cagagaaacg tgagatatat gatcaatatg gagaggacgc cctcaaggag 240
ggaatgggca gtggaggtgg tgctcatgac ccatttgaca ttttccaatc cttctttggt 300
ggcaacccat ttggtggtgg tggtagcagc agaggccgta ggaaggaggg agaggatgtt 360
atccatcctc tcaaggtttc tttggaagat ctctacaatg gcacctcaaa gaagctgtct 420
ctttcccgta atgttatctg ctcaaagtgc aaaggtaaag ggtccaaatc aggtgcatca 480
atgaaatgtt cgggttgcca aggttctgga atgaaggtct ccataagaca acttggtccc 540
tctatgatcc agcaaatgca gcatccttgt aatgaatgta agggtactgg tgagaccatt 600
aatgataagg atcgttgccc tcaatgtaaa ggtgaaaagg ttgttcagga gaagaaagtg 660
ctggaagtta ttgttgagaa gggtatgcaa aatggacaga ggattacttt ccctggagaa 720
gctgatgaag ctcctgatac tattacaggg gacattgttt ttgtccttca gcaaaaggag 780
catcctaagt tcaagcgaaa gggtgatgac ctaattgttg atcacacttt atctcttaca 840
gaggcacttt gtgcctccca gtttatatta acccatctag atggagacct cctcataaaa 900
tcccaacctg gggaggtagt gaagcctgat caattcaagg ccataaatga tgaagggatg 960
ccaatgtatc agaggccatt catgaggggg aaactgtaca ttcatttcag tgttgatttc 1020
ccagactctc tgccccctga tcagtgcaaa gccctagagg cagttcttcc ctcaagaaca 1080
tcagtccagc tgtctgacat ggagctggat gaatgtgagg agacaacttt acacgatgtg 1140
aactttgacg aggagatgcg aaggaagcaa caacaggccc aagaggcata tgatgaagat 1200
gatgatatgc atggtggtgg ccagagggtg caatgtgctc agcaataa 1248
<210>30
<211>415
<212>PRT
<213>橡胶
<400>30
Met Phe Gly Arg Ala Pro Lys Lys Ser Asp Asn Thr Lys Tyr Tyr Glu
1 5 10 15
Ile Leu Gly Val Ser Lys Asn Ala Ser Gln Asp Asp Leu Lys Lys Ala
20 25 30
Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Ser Gly Gly Gly Ala His Asp Pro Phe Asp Ile Phe Gln
85 90 95
Ser Phe Phe Gly Gly Asn Pro Phe Gly Gly Gly Gly Ser Ser Arg Gly
100 105 110
Arg Arg Lys Glu Gly Glu Asp Val Ile His Pro Leu Lys Val Ser Leu
115 120 125
Glu Asp Leu Tyr Asn Gly Thr Ser Lys Lys Leu Ser Leu Ser Arg Asn
130 135 140
Val Ile Cys Ser Lys Cys Lys Gly Lys Gly Ser Lys Ser Gly Ala Ser
145 150 155 160
Met Lys Cys Ser Gly Cys Gln Gly Ser Gly Met Lys Val Ser Ile Arg
165 170 175
Gln Leu Gly Pro Ser Met Ile Gln Gln Met Gln His Pro Cys Asn Glu
180 185 190
Cys Lys Gly Thr Gly Glu Thr Ile Asn Asp Lys Asp Arg Cys Pro Gln
195 200 205
Cys Lys Gly Glu Lys Val Val Gln Glu Lys Lys Val Leu Glu Val Ile
210 215 220
Val Glu Lys Gly Met Gln Asn Gly Gln Arg Ile Thr Phe Pro Gly Glu
225 230 235 240
Ala Asp Glu Ala Pro Asp Thr Ile Thr Gly Asp Ile Val Phe Val Leu
245 250 255
Gln Gln Lys Glu His Pro Lys Phe Lys Arg Lys Gly Asp Asp Leu Ile
260 265 270
Val Asp His Thr Leu Ser Leu Thr Glu Ala Leu Cys Ala Ser Gln Phe
275 280 285
Ile Leu Thr His Leu Asp Gly Asp Leu Leu Ile Lys Ser Gln Pro Gly
290 295 300
Glu Val Val Lys Pro Asp Gln Phe Lys Ala Ile Asn Asp Glu Gly Met
305 310 315 320
Pro Met Tyr Gln Arg Pro Phe Met Arg Gly Lys Leu Tyr Ile His Phe
325 330 335
Ser Val Asp Phe Pro Asp Ser Leu Pro Pro Asp Gln Cys Lys Ala Leu
340 345 350
Glu Ala Val Leu Pro Ser Arg Thr Ser Val Gln Leu Ser Asp Met Glu
355 360 365
Leu Asp Glu Cys Glu Glu Thr Thr Leu His Asp Val Asn Phe Asp Glu
370 375 380
Glu Met Arg Arg Lys Gln Gln Gln Ala Gln Glu Ala Tyr Asp Glu Asp
385 390 395 400
Asp Asp Met His Gly Gly Gly Gln Arg Val Gln Cys Ala Gln Gln
405 410 415
<210>31
<211>1260
<212>DNA
<213>番茄(Lycopersicum esculentum)
<400>31
atgtttggaa gagcaccgaa gaagagcgat aatacaaagt attatgagat cttaggagtt 60
cctaaggctg cttctcagga agatctcaaa aaggcttatc gtaaagctgc tatcaaaaat 120
caccctgata agggaggcga tcctgagaag tttaaagagc ttgctcaagc ttatgaggtt 180
ctgagtgacc cggagaagcg tgagatatat gatcagtatg gagaggatgc tctaaaggaa 240
ggaatgggtg gtggaggtgg tggacatgaa ccatttgata tatttcaatc attcttcggt 300
ggtggtggaa acccctttgg tggtggtgga agcagcagag tccgaagaca gagaagagga 360
gaggatgtta tccacccgct caaggtttct ttagaggatc tttacaatgg gacatcaaag 420
aagctttcac tatctcgcaa tgtgttgtgc tcaaagtgca agggcaaagg ttccaagtca 480
ggtgcttcaa tgaaatgttc tggctgtcaa gggtctggaa tgaaagtttc tatcagacag 540
ctcggtccat ccatgatcca gcagatgcag cacccttgca atgagtgcaa gggtactgga 600
gagacgatca gtgacaaaga taggtgccct cagtgcaagg gtgagaaggt tgtgcaggag 660
aagaaggtgt tggaagttca cgtggagaag ggtatgcaga atgggcaaaa gataacattt 720
ccaggcgagg cagatgaagc gccagatacc atcactggag acattgtttt tgtcttgcaa 780
caaaaggaac atcctaagtt caagcgaaag ggagatgatc tttttgttga gcacacattg 840
agccttgacg agtctctatg tggtttccag tttgttctga ctcacctaga caacagacag 900
ctgctcatta agtcccaacc tggcgaagtt gtcaagcctg atcagtttaa ggctatcaac 960
gatgaaggaa tgccgatgta ccaaaggccg ttcatgaagg gcaaaatgta cattcacttc 1020
actgttgatt tccccgagtc attacacgca gagcagtgca agaaccttga ggctgtgctg 1080
cctcccaaaa ccaaattgca gatatcagat atggaattgg acgagtggga ggagactact 1140
ttgcacgatg tcaacattga ggaggagatg cgaaggaagc agcaagctgc ccaagaggca 1200
caggacgaag atgacgatat gcctggtggt gcacagagag tccaatgtgc acagcagtaa 1260
<210>32
<211>419
<212>PRT
<213>番茄
<400>32
Met Phe Gly Arg Ala Pro Lys Lys Ser Asp Asn Thr Lys Tyr Tyr Glu
1 5 10 15
Ile Leu Gly Val Pro Lys Ala Ala Ser Gln Glu Asp Leu Lys Lys Ala
20 25 30
Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Gly Gly Gly Gly Gly His Glu Pro Phe Asp Ile Phe Gln
85 90 95
Ser Phe Phe Gly Gly Gly Gly Asn Pro Phe Gly Gly Gly Gly Ser Ser
100 105 110
Arg Val Arg Arg Gln Arg Arg Gly Glu Asp Val Ile His Pro Leu Lys
115 120 125
Val Ser Leu Glu Asp Leu Tyr Asn Gly Thr Ser Lys Lys Leu Ser Leu
130 135 140
Ser Arg Asn Val Leu Cys Ser Lys Cys Lys Gly Lys Gly Ser Lys Ser
145 150 155 160
Gly Ala Ser Met Lys Cys Ser Gly Cys Gln Gly Ser Gly Met Lys Val
165 170 175
Ser Ile Arg Gln Leu Gly Pro Ser Met Ile Gln Gln Met Gln His Pro
180 185 190
Cys Asn Glu Cys Lys Gly Thr Gly Glu Thr Ile Ser Asp Lys Asp Arg
195 200 205
Cys Pro Gln Cys Lys Gly Glu Lys Val Val Gln Glu Lys Lys Val Leu
210 215 220
Glu Val His Val Glu Lys Gly Met Gln Asn Gly Gln Lys Ile Thr Phe
225 230 235 240
Pro Gly Glu Ala Asp Glu Ala Pro Asp Thr Ile Thr Gly Asp Ile Val
245 250 255
Phe Val Leu Gln Gln Lys Glu His Pro Lys Phe Lys Arg Lys Gly Asp
260 265 270
Asp Leu Phe Val Glu His Thr Leu Ser Leu Asp Glu Ser Leu Cys Gly
275 280 285
Phe Gln Phe Val Leu Thr His Leu Asp Asn Arg Gln Leu Leu Ile Lys
290 295 300
Ser Gln Pro Gly Glu Val Val Lys Pro Asp Gln Phe Lys Ala Ile Asn
305 310 315 320
Asp Glu Gly Met Pro Met Tyr Gln Arg Pro Phe Met Lys Gly Lys Met
325 330 335
Tyr Ile His Phe Thr Val Asp Phe Pro Glu Ser Leu His Ala Glu Gln
340 345 350
Cys Lys Asn Leu Glu Ala Val Leu Pro Pro Lys Thr Lys Leu Gln Ile
355 360 365
Ser Asp Met Glu Leu Asp Glu Trp Glu Glu Thr Thr Leu His Asp Val
370 375 380
Asn Ile Glu Glu Glu Met Arg Arg Lys Gln Gln Ala Ala Gln Glu Ala
385 390 395 400
Gln Asp Glu Asp Asp Asp Met Pro Gly Gly Ala Gln Arg Val Gln Cys
405 410 415
Ala Gln Gln
<210>33
<211>1272
<212>DNA
<213>紫花苜蓿(Medicago sativa)
<400>33
atgtttgggc gcggaccaac aaggaagagt gataacacca aatattacga tattcttggt 60
gtttcaaaaa gtgctagtga agatgaaatc aagaaagcct atagaaaggc agcgatgaag 120
aaccatccag ataagggtgg ggatcctgag aagttcaagg agttgggcca agcatatgaa 180
gtgttgagcg atcctgaaaa gaaagaactg tatgatcaat atggtgaaga tgcccttaaa 240
gaaggaatgg ggggaggcgc aggaagctca tttcataatc cgtttgatat tttcgaatca 300
ttttttggtg caggctttgg tggtggtggt ccttcacgcg caagaagaca gaagcaagga 360
gaagatgtgg tgcattctat aaaggtttcc ttggaggatg tgtataacgg cactacaaag 420
aagctatcac tttctaggaa tgcactgtgc tcaaaatgta aagggaaagg ttcaaaaagt 480
ggaactgctg gaaggtgttt tggatgccag ggcacaggta tgaagattac cagaaggcaa 540
attggactgg gcatgattca acaaatgcaa cacgtctgtc ctgactgcaa aggaacaggc 600
gaggtcatta gtgagagaga tagatgccct caatgcaagg gaaacaagat tactcaagaa 660
aagaaggtgc tggaggtgca tgtggaaaag gggatgcagc agggtcacaa gattgtattc 720
gaaggacaag ctgatgaact ccctgataca atcacaggag acatagtttt tgtcttgcaa 780
gtaaagggac atccgaagtt tcggagggag cgtgatgacc ttcacattga acacaatttg 840
agcttaactg atgctctctg tggcttccag tttaatgtca cacatcttga tggaaggcaa 900
ctattggtca aatcgaaccc cggcgaagtc atcaagccag gtcaacataa agctataaat 960
gatgagggaa tgccacaaca tggtaggccg ttcatgaagg gacgcctata catcaagttt 1020
agtgttgatt tcccggattc gggttttctt tccccaagcc aaagcctgga attagaaaag 1080
atattacctc aaaagacaag caagaacttg tcccaaaagg aggtagatga ttgtgaggag 1140
accaccctgc atgatgtcaa tattgcagag gagatgagtc gaaagaagca acaataccgt 1200
gaggcatatg atgacgatga tgatgaagat gatgagcact cgcagcctcg ggtgcaatgc 1260
gctcaacagt ag 1272
<210>34
<211>423
<212>PRT
<213>紫花苜蓿
<400>34
Met Phe Gly Arg Gly Pro Thr Arg Lys Ser Asp ASh Thr Lys Tyr Tyr
1 5 10 15
Asp Ile Leu Gly Val Ser Lys Ser Ala Ser Glu Asp Glu Ile Lys Lys
20 25 30
Ala Tyr Arg Lys Ala Ala Met Lys Asn His Pro Asp Lys Gly Gly Asp
35 40 45
Pro Glu Lys Phe Lys Glu Leu Gly Gln Ala Tyr Glu Val Leu Ser Asp
50 55 60
Pro Glu Lys Lys Glu Leu Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys
65 70 75 80
Glu Gly Met Gly Gly Gly Ala Gly Ser Ser Phe His Asn Pro Phe Asp
85 90 95
Ile Phe Glu Ser Phe Phe Gly Ala Gly Phe Gly Gly Gly Gly Pro Ser
100 105 110
Arg Ala Arg Arg Gln Lys Gln Gly Glu Asp Val Val His Ser Ile Lys
115 120 125
Val Ser Leu Glu Asp Val Tyr Asn Gly Thr Thr Lys Lys Leu Ser Leu
130 135 140
Ser Arg Asn Ala Leu Cys Ser Lys Cys Lys Gly Lys Gly Ser Lys Ser
145 150 155 160
Gly Thr Ala Gly Arg Cys Phe Gly Cys Gln Gly Thr Gly Met Lys Ile
165 170 175
Thr Arg Arg Gln Ile Gly Leu Gly Met Ile Gln Gln Met Gln His Val
180 185 190
Cys Pro Asp Cys Lys Gly Thr Gly Glu Val Ile Ser Glu Arg Asp Arg
195 200 205
Cys Pro Gln Cys Lys Gly Asn Lys Ile Thr Gln Glu Lys Lys Val Leu
210 215 220
Glu Val His Val Glu Lys Gly Met Gln Gln Gly His Lys Ile Val Phe
225 230 235 240
Glu Gly Gln Ala Asp Glu Leu Pro Asp Thr Ile Thr Gly Asp Ile Val
245 250 255
Phe Val Leu Gln Val Lys Gly His Pro Lys Phe Arg Arg Glu Arg Asp
260 265 270
Asp Leu His Ile Glu His Asn Leu Ser Leu Thr Asp Ala Leu Cys Gly
275 280 285
Phe Gln Phe Asn Val Thr His Leu Asp Gly Arg Gln Leu Leu Val Lys
290 295 300
Ser Asn Pro Gly Glu Val Ile Lys Pro Gly Gln His Lys Ala Ile Asn
305 310 315 320
Asp Glu Gly Met Pro Gln His Gly Arg Pro Phe Met Lys Gly Arg Leu
325 330 335
Tyr Ile Lys Phe Ser Val Asp Phe Pro Asp Ser Gly Phe Leu Ser Pro
340 345 350
Ser Gln Ser Leu Glu Leu Glu Lys Ile Leu Pro Gln Lys Thr Ser Lys
355 360 365
Asn Leu Ser Gln Lys Glu Val Asp Asp Cys Glu Glu Thr Thr Leu His
370 375 380
Asp Val Asn Ile Ala Glu Glu Met Ser Arg Lys Lys Gln Gln Tyr Arg
385 390 395 400
Glu Ala Tyr Asp Asp Asp Asp Asp Glu Asp Asp Glu His Ser Gln Pro
405 410 415
Arg Val Gln Cys Ala Gln Gln
420
<210>35
<211>1257
<212>DNA
<213>烟草(Nicotiana tabacum)
<400>35
atgtttggga gaggaccaaa gaagagtgat aatacgaggt actatgaaat attgggtgtg 60
tcaaagaatg catcagatga tgaaatcaag aaagcttata gaaaagctgc tatgaagaat 120
caccctgata agggtggtga ccctgaaaag tttaaggagc ttgctcaagc ttatgaggtg 180
ttgagtgact cacagaagcg tgagatttat gatcagtatg gagaagatgc attaaaagaa 240
ggaatgggtg gcggcggcgg aatgcatgat ccatttgaca tctttgaatc tttctttggt 300
ggcaatccat ttggaggtgg tggtagcagc agaggaagaa gacagagaag gggtgaggat 360
gtagtgcatc cactgaaggt ctctctcgag gacctttaca gtgggataac caaaaaactc 420
tccctttcgc gcaatgtcat ttgctccaag tgcagtggga aaggatcgaa gtctggtgct 480
tcaatgaagt gttctggttg taaaggtagt ggtatgaagg tttcaattag acaacttggc 540
ccttcaatga tccagcaaat gcagcacgct tgtaatgaat gcaagggtac tggagagact 600
attgacgata aggatcggtg ccctcggtgc aaaggtgaaa aagtggttca ggagaagaaa 660
gtccttgaag ttcatgttga gaaaggcatg caaaatggac agaaaattac attccctgga 720
aaggctgatg aaacccctga tgcaattact ggagatatag tttttgtgct ccagcagaaa 780
gacacccgag gttccaagag aaagggcgac gatctgtttg tagatcacac attgagtcta 840
actgaggctt tatgtggctt ccagttcata atgacacact tggatggcag acaactcctc 900
ataaaatcaa atctcgggga agttgttaaa cctgatcaat tcaaggcaat caatgatgag 960
ggaacgccaa tgtatcagag gccatttatg aggggcaaat tgtacattcg tttcgtcgtt 1020
gaattcccag attcattgaa cacagaacag gtgaaggctc tggaggcaat cttaccacca 1080
agacctcagt cacagtacac agacatggaa ttggatgagt gtgaggagac ttctttacat 1140
gatgtgaata ttgaggagga aatgagaagg aaacaggcag ctcaacaaga ggcatatgat 1200
gaggatgatg agatgcatgg tggtggagga cagagagtac aatgtgcaca gcagtaa 1257
<210>36
<211>418
<212>PRT
<213>烟草
<400>36
Met Phe Gly Arg Gly Pro Lys Lys Ser Asp Asn Thr Arg Tyr Tyr Glu
1 5 10 15
Ile Leu Gly Val Ser Lys Asn Ala Ser Asp Asp Glu Ile Lys Lys Ala
20 25 30
Tyr Arg Lys Ala Ala Met Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Ser
50 55 60
Gln Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Gly Gly Gly Gly Met His Asp Pro Phe Asp Ile Phe Glu
85 90 95
Ser Phe Phe Gly Gly Asn Pro Phe Gly Gly Gly Gly Ser Ser Arg Gly
100 105 110
Arg Arg Gln Arg Arg Gly Glu Asp Val Val His Pro Leu Lys Val Ser
115 120 125
Leu Glu Asp Leu Tyr Ser Gly Ile Thr Lys Lys Leu Ser Leu Ser Arg
130 135 140
Asn Val Ile Cys Ser Lys Cys Ser Gly Lys Gly Ser Lys Ser Gly Ala
145 150 155 160
Ser Met Lys Cys Ser Gly Cys Lys Gly Ser Gly Met Lys Val Ser Ile
165 170 175
Arg Gln Leu Gly Pro Ser Met Ile Gln Gln Met Gln His Ala Cys Asn
180 185 190
Glu Cys Lys Gly Thr Gly Glu Thr Ile Asp Asp Lys Asp Arg Cys Pro
195 200 205
Arg Cys Lys Gly Glu Lys Val Val Gln Glu Lys Lys Val Leu Glu Val
210 215 220
His Val Glu Lys Gly Met Gln Asn Gly Gln Lys Ile Thr Phe Pro Gly
225 230 235 240
Lys Ala Asp Glu Thr Pro Asp Ala Ile Thr Gly Asp Ile Val Phe Val
245 250 255
Leu Gln Gln Lys Asp Thr Arg Gly Ser Lys Arg Lys Gly Asp Asp Leu
260 265 270
Phe Val Asp His Thr Leu Ser Leu Thr Glu Ala Leu Cys Gly Phe Gln
275 280 285
Phe Ile Met Thr His Leu Asp Gly Arg Gln Leu Leu Ile Lys Ser Asn
290 295 300
Leu Gly Glu Val Val Lys Pro Asp Gln Phe Lys Ala Ile Asn Asp Glu
305 310 315 320
Gly Thr Pro Met Tyr Gln Arg Pro Phe Met Arg Gly Lys Leu Tyr Ile
325 330 335
Arg Phe Val Val Glu Phe Pro Asp Ser Leu Asn Thr Glu Gln Val Lys
340 345 350
Ala Leu Glu Ala Ile Leu Pro Pro Arg Pro Gln Ser Gln Tyr Thr Asp
355 360 365
Met Glu Leu Asp Glu Cys Glu Glu Thr Ser Leu His Asp Val Asn Ile
370 375 380
Glu Glu Glu Met Arg Arg Lys Gln Ala Ala Gln Gln Glu Ala Tyr Asp
385 390 395 400
Glu Asp Asp Glu Met His Gly Gly Gly Gly Gln Arg Val Gln Cys Ala
405 410 415
Gln Gln
<210>37
<211>1272
<212>DNA
<213>河柳(Salix gilgiana)
<400>37
atgtttgggc gtgctccgag gaggagtgac aacaccaagt attatgaggt tttggctgtg 60
tcaaaaggtg caagtcaaga tgaactgaag aaggcttata agaaagctgc cataaagaat 120
catcctgata aaggtggaga tcctgaaaag ttcaaggagt tgtctcaagc ttatgaagtc 180
cttagtgatc cagataaaag agaaatttat gatcaatatg gggaagatgc acttaaggag 240
gggatgggac ctggtggtgg tgggggtggt cacaatccat ttgatatatt cgaatcattt 300
tttggtggag gtggttttgg tggtggtagc agctcaagag gaagaaggca gaagcaaggt 360
gaagatgtag cgcaccctct gaaggtttcc ttagaggatt tgtacaatgg aacttcaaag 420
aaactctctc tttccagaaa cattttgtgt gccaaatgta aagggaaagg ttcaaagagt 480
ggagcctttg ggaaatgtcg tggctgccaa ggtactggaa tgaaagtttc aatccgacaa 540
attggattgg gcatgatgca acaaatgcaa catgtgtgtc ctgaatgcag gggctcaggt 600
gagctaatta gtgagaagga taaatgccct cattgcagag ggaacaaggt aacgcaggaa 660
aagagggtgc tggaagtgca tgttgaaagg ggaatgcagc atggccagaa gatagttttc 720
gaaggtcaag ctgatgaagc tcctgacaca attacagggg atgttgtttt tgtattgcaa 780
gaaaaagc actccaagtt tgaacggaaa atggatgatc tctttgtgga acactctctc 840
agtttaacag aggctctttg cgggtatcag tttgccctta cccatcttga tggtcggcag 900
cttcttatca aatcaaatcc ttacgagatt gtaaaacctg gtcaatacaa agcaattaac 960
gatgaaggaa tgccacatca tcacaggccc ttcatgaggg gcaagctcta tatccatttt 1020
aatgtggtgt tccctgactc gggcactcta tcccctgagc agtgccgtac tttagagact 1080
atactacccc caaggcaaag caaaaacttg tcagagatgg agattgataa ctgcgaagag 1140
acaattatgc atgatgtcaa tatggaggag gagaaaaggc ggaaacagca gcagcgccac 1200
cagcatgaag catatgatga ggatgaggag gaggaatcat ccatgccccg ggtgcagtgt 1260
gcccagcagt aa 1272
<210>38
<211>423
<212>PRT
<213>河柳
<400>38
Met Phe Gly Arg Ala Pro Arg Arg Ser Asp Asn Thr Lys Tyr Tyr Glu
1 5 10 15
Val Leu Ala Val Ser Lys Gly Ala Ser Gln Asp Glu Leu Lys Lys Ala
20 25 30
Tyr Lys Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ser Gln Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Asp Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Pro Gly Gly Gly Gly Gly Gly His Asn Pro Phe Asp Ile
85 90 95
Phe Glu Ser Phe Phe Gly Gly Gly Gly Phe Gly Gly Gly Ser Ser Ser
100 105 110
Arg Gly Arg Arg Gln Lys Gln Gly Glu Asp Val Ala His Pro Leu Lys
115 120 125
Val Ser Leu Glu Asp Leu Tyr Asn Gly Thr Ser Lys Lys Leu Ser Leu
130 135 140
Ser Arg Ash Ile Leu Cys Ala Lys Cys Lys Gly Lys Gly Ser Lys Ser
145 150 155 160
Gly Ala Phe Gly Lys Cys Arg Gly Cys Gln Gly Thr Gly Met Lys Val
165 170 175
Ser Ile Arg Gln Ile Gly Leu Gly Met Met Gln Gln Met Gln His Val
180 185 190
Cys Pro Glu Cys Arg Gly Ser Gly Glu Leu Ile Ser Glu Lys Asp Lys
195 200 205
Cys Pro His Cys Arg Gly Asn Lys Val Thr Gln Glu Lys Arg Val Leu
210 215 220
Glu Val His Val Glu Arg Gly Met Gln His Gly Gln Lys Ile Val Phe
225 230 235 240
Glu Gly Gln Ala Asp Glu Ala Pro Asp Thr Ile Thr Gly Asp Val Val
245 250 255
Phe Val Leu Gln Leu Lys Lys His Ser Lys Phe Glu Arg Lys Met Asp
260 265 270
Asp Leu Phe Val Glu His Ser Leu Ser Leu Thr Glu Ala Leu Cys Gly
275 280 285
Tyr Gln Phe Ala Leu Thr His Leu Asp Gly Arg Gln Leu Leu Ile Lys
290 295 300
Ser Asn Pro Tyr Glu Ile Val Lys Pro Gly Gln Tyr Lys Ala Ile Asn
305 310 315 320
Asp Glu Gly Met Pro His His His Arg Pro Phe Met Arg Gly Lys Leu
325 330 335
Tyr Ile His Phe Asn Val Val Phe Pro Asp Ser Gly Thr Leu Ser Pro
340 345 350
Glu Gln Cys Arg Thr Leu Glu Thr Ile Leu Pro Pro Arg Gln Ser Lys
355 360 365
Asn Leu Ser Glu Met Glu Ile Asp Asn Cys Glu Glu Thr Ile Met His
370 375 380
Asp Val Asn Met Glu Glu Glu Lys Arg Arg Lys Gln Gln Gln Arg His
385 390 395 400
Gln His Glu Ala Tyr Asp Glu Asp Glu Glu Glu Glu Ser Ser Met Pro
405 410 415
Arg Val Gln Cys Ala Gln Gln
420
<210>39
<211>1263
<212>DNA
<213>河柳
<400>39
atgtttggga gagcaccaaa gaaaagcgac aacaccaagt actatgaggt tcttggagtc 60
tcaaagagtg cttcacagga tgatctaaag aaggcttata ggaaagcagc tatcaagaac 120
catcctgata agggcggtga tcctgaaaag ttcaaggagt tggcgcaagc atatgaggtt 180
ctgagtgacc ctgagaagcg tgagatatat gatcagtatg gagaggatgc cctcaaggaa 240
ggaatgggta gcggcggcag cggcgctcac gatccattcg atatcttcca atccttcttt 300
ggtggtggca atccattcgg tggtggaggt agcagcaggg gccgaaggca aagaaggggc 360
gaggatgtga tccaccctct gaaagtttct tttgaagacc tttataatgg cacatccaag 420
aagctttctc tttcacgaaa tgtaatctgc tccaagtgca agggcaaagg ttccaaatcc 480
ggagcatcat caaaatgtgc tggttgccaa ggttctggaa tgaaggtctc cataagacac 540
ctcggtcctt ctatgatcca gcaaatgcag catgcctgca atgaatgcaa gggcactggc 600
gagacaatta acgataagga ccgatgccct caatgcaagg gtgagaaggt tgtccaggag 660
aagaaagtgt tggaagtagt tgttgagaag ggcatgcaaa atgggcagaa ggtaacattt 720
cctggagaag ctgatgaggc gcctgacact gttacagggg acatagtctt cgtcctgcag 780
caaaaggatc accctaagtt taagagaaag ggtgatgacc tatttgttga gcacacacta 840
tctcttactg aggcactatg tggcttccaa ttcgtcttga cccatttgga tggaaggcag 900
ctcctgataa aatctcaacc cggggaagta gtcaagcctg atcaattcaa ggctataaat 960
gatgaaggaa tgccgatgta ccaaaggcca tttatgagag ggaaactcta cattcatttc 1020
agtgttgaat tcccagactc cctgtcccct gatatgtgca aggcgttgga ggccgtgctt 1080
cctccgcgag cctctgttca gctgactgac atggagcttg atgaatgcga ggaaactact 1140
ttacatgatg tgaacatcga tgaggagatg aggaggaaac agcaacagca ggcccaagaa 1200
gcgtatgatg aagatgatga gatgcctggt ggtgcccaga gggtgcagtg tgctcagcaa 1260
taa 1263
<210>40
<211>420
<212>PRT
<213>河柳
<400>40
Met Phe Gly Arg Ala Pro Lys Lys Ser Asp Asn Thr Lys Tyr Tyr Glu
1 5 10 15
Val Leu Gly Val Ser Lys Ser Ala Ser Gln Asp Asp Leu Lys Lys Ala
20 25 30
Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Ser Gly Gly Ser Gly Ala His Asp Pro Phe Asp Ile Phe
85 90 95
Gln Ser Phe Phe Gly Gly Gly Asn Pro Phe Gly Gly Gly Gly Ser Ser
100 105 110
Arg Gly Arg Arg Gln Arg Arg Gly Glu Asp Val Ile His Pro Leu Lys
115 120 125
Val Ser Phe Glu Asp Leu Tyr Asn Gly Thr Ser Lys Lys Leu Ser Leu
130 135 140
Ser Arg Asn Val Ile Cys Ser Lys Cys Lys Gly Lys Gly Ser Lys Ser
145 150 155 160
Gly Ala Ser Ser Lys Cys Ala Gly Cys Gln Gly Ser Gly Met Lys Val
165 170 175
Ser Ile Arg His Leu Gly Pro Ser Met Ile Gln Gln Met Gln His Ala
180 185 190
Cys Asn Glu Cys Lys Gly Thr Gly Glu Thr Ile Asn Asp Lys Asp Arg
195 200 205
Cys Pro Gln Cys Lys Gly Glu Lys Val Val Gln Glu Lys Lys Val Leu
210 215 220
Glu Val Val Val Glu Lys Gly Met Gln Asn Gly Gln Lys Val Thr Phe
225 230 235 240
Pro Gly Glu Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Val
245 250 255
Phe Val Leu Gln Gln Lys Asp His Pro Lys Phe Lys Arg Lys Gly Asp
260 265 270
Asp Leu Phe Val Glu His Thr Leu Ser Leu Thr Glu Ala Leu Cys Gly
275 280 285
Phe Gln Phe Val Leu Thr His Leu Asp Gly Arg Gln Leu Leu Ile Lys
290 295 300
Ser Gln Pro Gly Glu Val Val Lys Pro Asp Gln Phe Lys Ala Ile Asn
305 310 315 320
Asp Glu Gly Met Pro Met Tyr Gln Arg Pro Phe Met Arg Gly Lys Leu
325 330 335
Tyr Ile His Phe Ser Val Glu Phe Pro Asp Ser Leu Ser Pro Asp Met
340 345 350
Cys Lys Ala Leu Glu Ala Val Leu Pro Pro Arg Ala Ser Val Gln Leu
355 360 365
Thr Asp Met Glu Leu Asp Glu Cys Glu Glu Thr Thr Leu His Asp Val
370 375 380
Asn Ile Asp Glu Glu Met Arg Arg Lys Gln Gln Gln Gln Ala Gln Glu
385 390 395 400
Ala Tyr Asp Glu Asp Asp Glu Met Pro Gly Gly Ala Gln Arg Val Gln
405 410 415
Cys Ala Gln Gln
420
<210>41
<211>1260
<212>DNA
<213>马铃薯(Solanum tuberosum)
<400>41
atgtttggga gggcaccaga gaagagcgac aacacgaagt actatgagat cttaggtgtc 60
cctaagactg ctgcacagga agatctcaag aaagcttacc gtaaagctgc tattaagaat 120
catcctgata agggaggtga tcctgaaaag tttaaagagc ttgcacaagc ttatgaggtt 180
ctgagtgatc ccgagaagcg tgagatatat gatcagtatg gagaagatgc tctcaaggaa 240
ggaatgggtg gtggaggtgg tggacacgac ccatttgata ttttctcatc tttctttggt 300
ggcagcccat ttggtggagg tggtggaagc agcagaggaa gaagacaaag aagaggagag 360
gatgttgtcc atcctctcaa agtttctctg gaggatctgt acaatggaac atcaaagaag 420
ctgtcactat ctcgcaatgt attgtgctcg aagtgcaagg ggaaaggatc taaatcaggt 480
gcttcaatga agtgttctgg ctgtcaaggg tctgggatga aagtcactat tagacaactt 540
ggcccatcca tgatccagca gatgcagcac ccttgcaacg agtgtaaggg tactggtgag 600
atgatcaatg ataaagatag gtgtgggcag tgtaaaggtg agaaagttgt gcaggagaag 660
aaggtgttgg aagttgttgt cgagaagggt atgcagaacg gacagaagat aacattcccg 720
ggcgaagctg atgaagcacc tgataccgtc actggggaca tagtttttgt cttgcaacag 780
aaggaacatc ccaagtttaa gcgaaaggga gatgatctct ttgtagagca caccttgagc 840
ttaaccgagg ccctgtgtgg tttccagttc atcttgactc acctagataa taggcagctg 900
atcatcaagc cccaagccgg agaagttgtc aagcctgatc aatttaaagc cataaatgat 960
gaaggaatgc ctatgtacca aaggccattt atgagaggaa aactatacat tcactttact 1020
gtagaattcc ccgacacatt atcccccgag caatgcaaga accttgaagc agtattgcca 1080
ccaaaaccga aaacacaaat gactgatatg gaattggacg agtgcgagga gaccacctta 1140
catgatgtta acatcgaaga ggagatgcgg aggaagcagc aacaggccca agaggcatat 1200
gacgaagatg atgaagacat gcatggaggt gcacagagag ttcagtgtgc acaacagtaa 1260
<210>42
<211>419
<212>PRT
<213>马铃薯
<400>42
Met Phe Gly Arg Ala Pro Glu Lys Ser Asp Asn Thr Lys Tyr Tyr Glu
1 5 10 15
Ile Leu Gly Val Pro Lys Thr Ala Ala Gln Glu Asp Leu Lys Lys Ala
20 25 30
Tyr Arg Lys Ala Ala Ile Lys Asn His Pro Asp Lys Gly Gly Asp Pro
35 40 45
Glu Lys Phe Lys Glu Leu Ala Gln Ala Tyr Glu Val Leu Ser Asp Pro
50 55 60
Glu Lys Arg Glu Ile Tyr Asp Gln Tyr Gly Glu Asp Ala Leu Lys Glu
65 70 75 80
Gly Met Gly Gly Gly Gly Gly Gly His Asp Pro Phe Asp Ile Phe Ser
85 90 95
Ser Phe Phe Gly Gly Ser Pro Phe Gly Gly Gly GIy Gly Ser Ser Arg
100 105 110
Gly Arg Arg Gln Arg Arg Gly Glu Asp Val Val His Pro Leu Lys Val
115 120 125
Ser Leu Glu Asp Leu Tyr Asn Gly Thr Ser Lys Lys Leu Ser Leu Ser
130 135 140
Arg Asn Val Leu Cys Ser Lys Cys Lys Gly Lys Gly Ser Lys Ser Gly
145 150 155 160
Ala Ser Met Lys Cys Ser Gly Cys Gln Gly Ser Gly Met Lys Val Thr
165 170 175
Ile Arg Gln Leu Gly Pro Ser Met Ile Gln Gln Met Gln His Pro Cys
180 185 190
Asn Glu Cys Lys Gly Thr Gly Glu Met Ile Asn Asp Lys Asp Arg Cys
195 200 205
Gly Gln Cys Lys Gly Glu Lys Val Val Gln Glu Lys Lys Val Leu Glu
210 215 220
Val Val Val Glu Lys Gly Met Gln Asn Gly Gln Lys Ile Thr Phe Pro
225 230 235 240
Gly Glu Ala Asp Glu Ala Pro Asp Thr Val Thr Gly Asp Ile Val Phe
245 250 255
Val Leu Gln Gln Lys Glu His Pro Lys Phe Lys Arg Lys Gly Asp Asp
260 265 270
Leu Phe Val Glu His Thr Leu Ser Leu Thr Glu Ala Leu Cys Gly Phe
275 280 285
Gln Phe Ile Leu Thr His Leu Asp Asn Arg Gln Leu Ile Ile Lys Pro
290 295 300
Gln Ala Gly Glu Val Val Lys Pro Asp Gln Phe Lys Ala Ile Asn Asp
305 310 315 320
Glu Gly Met Pro Met Tyr Gln Arg Pro Phe Met Arg Gly Lys Leu Tyr
325 330 335
Ile His Phe Thr Val Glu Phe Pro Asp Thr Leu Ser Pro Glu Gln Cys
340 345 350
Lys Asn Leu Glu Ala Val Leu Pro Pro Lys Pro Lys Thr Gln Met Thr
355 360 365
Asp Met Glu Leu Asp Glu Cys Glu Glu Thr Thr Leu His Asp Val Asn
370 375 380
Ile Glu Glu Glu Met Arg Arg Lys Gln Gln Gln Ala Gln Glu Ala Tyr
385 390 395 400
Asp Glu Asp Asp Glu Asp Met His Gly Gly Ala Gln Arg Val Gln Cys
405 410 415
Ala Gln Gln
<210>43
<211>1293
<212>DNA
<213>稻
<400>43
atggtcaagg acaccaaatt ctacgacacc ctcggctgcg cgcccgacgc taccgagtct 60
cagctcaaga ccgcataccg caagggcgcc ctcaagcacc accccgacaa gaacgcacac 120
tcgcccgaat ccgaggagaa gttcaaggag atctcacacg catacgaagt cctctcagac 180
ccccaaaagc gccaaatcta cgaccagtat ggtgaggagg gtctcgagca gggtggtgga 240
atgggcggcg gcggaggcat ggctgccgag gacttgttcg cacagttctt cggcggcggc 300
ggtggaggcg gaggcttcgg tggcatgggc ggcatgttcg gcggtcgcga gcccggcccc 360
aagaaggctc gcaccatcca ccacgttcac aaggtctctc tcgaggacat ctaccgcggc 420
aaggtttcca agcttgccct gcagaagagc gtcatctgct ccaagtgtga tggccgcggt 480
ggtaaggagg gtgctgtgaa gacttgccag ggctgccagg gccagggtat gaagaccatg 540
atgcgccaga tgggtcccat gatccagcga ttccagaccg tctgccccga ctgcaacggt 600
gagggggagc aggtccgcga gaaggacaag tgcaagcagt gctccggaaa gaagaccatc 660
atcgagcgca aggtgctcca cgtccacgtc gacaagggtg tgcaaagcgg caccaagatc 720
gacttcagag gcgagggtga ccagatgcct ggcgttgagc ccggtgatgt gcagttcgag 780
atcgagcaga agcctcaccc tcgcttccag cgcaagggtg acgacctcta ctaccacgcc 840
gagatcgacc ttcttactgc gctcgccggc ggtgccatct acgttgagca ccttgacgag 900
cgctggttga ccgtcgagat cctgcccggc gaggttatcg caccaggcga ggtcaaggtc 960
atccgcggcc agggtatgcc ctcataccgc caccacgacc acggcaacct ttacatccag 1020
ttcgacgtca agttccccac atccatccaa ggccctgccg acaaggacgg ccagtccacc 1080
tccatgtccg cacaacagat caaggccctc gaatccgtcc ttcctcctcg caagccccaa 1140
tcgatccctc ctcccgatgc tatgaccgag gacttccagc tcgagcgcgt agaccccatg 1200
gagggctccc gctccaaggg cgcccacagc atggacgagg acgatgacga gatgggcggc 1260
ggtggcgagc gcgtgcagtg cgcgtcgcag taa 1293
<210>44
<211>430
<212>PRT
<213>稻
<400>44
Met Val Lys Asp Thr Lys Phe Tyr Asp Thr Leu Gly Cys Ala Pro Asp
1 5 10 15
Ala Thr Glu Ser Gln Leu Lys Thr Ala Tyr Arg Lys Gly Ala Leu Lys
20 25 30
His His Pro Asp Lys Asn Ala His Ser Pro Glu Ser Glu Glu Lys Phe
35 40 45
Lys Glu Ile Ser His Ala Tyr Glu Val Leu Ser Asp Pro Gln Lys Arg
50 55 60
Gln Ile Tyr Asp Gln Tyr Gly Glu Glu Gly Leu Glu Gln Gly Gly Gly
65 70 75 80
Met Gly Gly Gly Gly Gly Met Ala Ala Glu Asp Leu Phe Ala Gln Phe
85 90 95
Phe Gly Gly Gly Gly Gly Gly Gly Gly Phe Gly Gly Met Gly Gly Met
100 105 110
Phe Gly Gly Arg Glu Pro Gly Pro Lys Lys Ala Arg Thr Ile His His
115 120 125
Val His Lys Val Ser Leu Glu Asp Ile Tyr Arg Gly Lys Val Ser Lys
130 135 140
Leu Ala Leu Gln Lys Ser Val Ile Cys Ser Lys Cys Asp Gly Arg Gly
145 150 155 160
Gly Lys Glu Gly Ala Val Lys Thr Cys Gln Gly Cys Gln Gly Gln Gly
165 170 175
Met Lys Thr Met Met Arg Gln Met Gly Pro Met Ile Gln Arg Phe Gln
180 185 190
Thr Val Cys Pro Asp Cys Asn Gly Glu Gly Glu Gln Val Arg Glu Lys
195 200 205
Asp Lys Cys Lys Gln Cys Ser Gly Lys Lys Thr Ile Ile Glu Arg Lys
210 215 220
Val Leu His Val His Val Asp Lys Gly Val Gln Ser Gly Thr Lys Ile
225 230 235 240
Asp Phe Arg Gly Glu Gly Asp Gln Met Pro Gly Val Glu Pro Gly Asp
245 250 255
Val Gln Phe Glu Ile Glu Gln Lys Pro His Pro Arg Phe Gln Arg Lys
260 265 270
Gly Asp Asp Leu Tyr Tyr His Ala Glu Ile Asp Leu Leu Thr Ala Leu
275 280 285
Ala Gly Gly Ala Ile Tyr Val Glu His Leu Asp Glu Arg Trp Leu Thr
290 295 300
Val Glu Ile Leu Pro Gly Glu Val Ile Ala Pro Gly Glu Val Lys Val
305 310 315 320
Ile Arg Gly Gln Gly Met Pro Ser Tyr Arg His His Asp His Gly Asn
325 330 335
Leu Tyr Ile Gln Phe Asp Val Lys Phe Pro Thr Ser Ile Gln Gly Pro
340 345 350
Ala Asp Lys Asp Gly Gln Ser Thr Ser Met Ser Ala Gln Gln Ile Lys
355 360 365
Ala Leu Glu Ser Val Leu Pro Pro Arg Lys Pro Gln Ser Ile Pro Pro
370 375 380
Pro Asp Ala Met Thr Glu Asp Phe Gln Leu Glu Arg Val Asp Pro Met
385 390 395 400
Glu Gly Ser Arg Ser Lys Gly Ala His Ser Met Asp Glu Asp Asp Asp
405 410 415
Glu Met Gly Gly Gly Gly Glu Arg Val Gln Cys Ala Ser Gln
420 425 430
<210>45
<211>1263
<212>DNA
<213>小麦
<400>45
atggtaaaag ataccaaact atatgatact ctgggtattt ccccgacctg tactgaagcc 60
gagttaaaaa aagcatacaa aatcggagca cttaaacacc atcctgataa aaacgcctca 120
aatccagccg ccgcagaaaa atttaaagaa atatcgcacg catatgaagt actatctgac 180
cctcaaaaaa gacacatata cgaccaatat ggcgaagagg gccttgaggg aggtggtggt 240
gctgcgggag ggatgaacgc agaagattta ttctctcaat tcttcagcgg tggctctgcc 300
ttcggaggtg gaggattggg tggcatgttc gggggagggc cacagcaacg tggcccccca 360
aaagcccgca ccattcatca cgttcacaag gtatctctag aagatatcta ccgcggtaaa 420
atctcaaaac tggcactaca aaagtcagtc atatgccaca agtgtgaggg acggggtggc 480
aaagatggtg cagtaaaaaa atgtgccggc tgtgatggac atggaatgaa aacaatgatg 540
cgtcaaatgg gtcctatgat tcagcggttt caaactcact gccccgactg caatggtgag 600
ggagaagtca tccgagagaa agataaatgt aagacgtgta acggtaaaaa gaccaacgtg 660
gaacgcaaag tactccacgt tcatgtggac agaggtgttc gatcggggca ccggattgaa 720
tttaaaggtg aaggagacca aacccccgga gttcaacctg gagatgttat ctttgaaatt 780
gagcagaaac cacatccaag attccaacga aaagacgatg accttattta ccacgcagag 840
atcgaccttg ttactgcctt agcgggcggg tcaatcttca ttgagcactt agacgaaaga 900
tggctgagtg tggagatact tcctggagag gttatctcac ctggatccgt taagatgata 960
cgcggtcagg gtatgccatc ccatcgtcac cacgactatg gaaatatgtt tgtacagttt 1020
gatgtcaaat tccccgaaag taactttgct gcaaattccg aggcatacgc agctctgaag 1080
agtattattc cgccgactgt ggtacctatc actccaccca ctgataccat gactgaaact 1140
gtatacttcg aagacattga ccctactcaa caagctcgtg cacagggtgc gacagcaatg 1200
gatgaagacg atgaagatgg ccatccagcc ggcgccgaac gggttcaatg tgcgtcacag 1260
taa 1263
<210>46
<211>420
<212>PRT
<213>小麦
<400>46
Met Val Lys Asp Thr Lys Leu Tyr Asp Thr Leu Gly Ile Ser Pro Thr
1 5 10 15
Cys Thr Glu Ala Glu Leu Lys Lys Ala Tyr Lys Ile Gly Ala Leu Lys
20 25 30
His His Pro Asp Lys Asn Ala Ser Asn Pro Ala Ala Ala Glu Lys Phe
35 40 45
Lys Glu Ile Ser His Ala Tyr Glu Val Leu Ser Asp Pro Gln Lys Arg
50 55 60
His Ile Tyr Asp Gln Tyr Gly Glu Glu Gly Leu Glu Gly Gly Gly Gly
65 70 75 80
Ala Ala Gly Gly Met Asn Ala Glu Asp Leu Phe Ser Gln Phe Phe Ser
85 90 95
Gly Gly Ser Ala Phe Gly Gly Gly Gly Leu Gly Gly Met Phe Gly Gly
100 105 110
Gly Pro Gln Gln Arg Gly Pro Pro Lys Ala Arg Thr Ile His His Val
115 120 125
His Lys Val Ser Leu Glu Asp Ile Tyr Arg Gly Lys Ile Ser Lys Leu
130 135 140
Ala Leu Gln Lys Ser Val Ile Cys His Lys Cys Glu Gly Arg Gly Gly
145 150 155 160
Lys Asp Gly Ala Val Lys Lys Cys Ala Gly Cys Asp Gly His Gly Met
165 170 175
Lys Thr Met Met Arg Gln Met Gly Pro Met Ile Gln Arg Phe Gln Thr
180 185 190
His Cys Pro Asp Cys Asn Gly Glu Gly Glu Val Ile Arg Glu Lys Asp
195 200 205
Lys Cys Lys Thr Cys Asn Gly Lys Lys Thr Asn Val Glu Arg Lys Val
210 215 220
Leu His Val His Val Asp Arg Gly Val Arg Ser Gly His Arg Ile Glu
225 230 235 240
Phe Lys Gly Glu Gly Asp Gln Thr Pro Gly Val Gln Pro Gly Asp Val
245 250 255
Ile Phe Glu Ile Glu Gln Lys Pro His Pro Arg Phe Gln Arg Lys Asp
260 265 270
Asp Asp Leu Ile Tyr His Ala Glu Ile Asp Leu Val Thr Ala Leu Ala
275 280 285
Gly Gly Ser Ile Phe Ile Glu His Leu Asp Glu Arg Trp Leu Ser Val
290 295 300
Glu Ile Leu Pro Gly Glu Val Ile Ser Pro Gly Ser Val Lys Met Ile
305 310 315 320
Arg Gly Gln Gly Met Pro Ser His Arg His His Asp Tyr Gly Asn Met
325 330 335
Phe Val Gln Phe Asp Val Lys Phe Pro Glu Ser Asn Phe Ala Ala Asn
340 345 350
Ser Glu Ala Tyr Ala Ala Leu Lys Ser Ile Ile Pro Pro Thr Val Val
355 360 365
Pro Ile Thr Pro Pro Thr Asp Thr Met Thr Glu Thr Val Tyr Phe Glu
370 375 380
Asp Ile Asp Pro Thr Gln Gln Ala Arg Ala Gln Gly Ala Thr Ala Met
385 390 395 400
Asp Glu Asp Asp Glu Asp Gly His Pro Ala Gly Ala Glu Arg Val Gln
405 410 415
Cys Ala Ser Gln
420
<210>47
<211>1320
<212>DNA
<213>秀丽新小杆线虫(Caenorhabditis elegans)
<400>47
atgtttggag gtggaagtag tggtcccgtg gacaccactt tatacacaac actcaatgtg 60
agaccagacg cttcgcaggc cgacattaag aaatcttact tcaaacttgc taaagaatac 120
catccagata aaaacccgga ccatggagat aaattcaaag agatcagttt tgcctatgaa 180
gttctttcga gccctgaaaa acgacgcttg tatgacgcca gaggtttgga aggagttcaa 240
ggaggaggag ctggtggtgg tggaggaggc tttcctggag gtctgttctc tcacttcttc 300
ggcggtgctg gcggtgatga cgatgacgac gatgatgata tgggtggtca tccatttggt 360
ggcttgttcg gtggaatggg tggaatggga cgaggtggcc cacgtcggcg gaaattccaa 420
gatactgttc atcccctcaa tgttacactc gaagagcttt acgtcggaaa aacatcaaag 480
ctgaagcttt ccaaaaaggc actctgtaaa acttgcgaag ggtcaggtgg aaagaaggga 540
gaaaaatata agtgtgatgc atgccgtggt cgtggagtga agacgatcgt tcagcaaatt 600
ggccccggaa tgctccaaca aatgcaggtt cactgtgatg cttgtaaggg ttctggaggc 660
aaagttccag caggtgataa gtgcaaagga tgccatggag aaaagtacga aaacgtttcg 720
aaaatattgg aggttcacgt tcttcctggc atgaaacata acgataaaat tacattcaaa 780
ggagatggag accaatctga cccagatggt gagccaggag atgttgtcat tgttattcaa 840
cagaaagatc atgatatttt caagagagat ggagatgatc ttcacatgac caagaaacta 900
tcactgaatg aggcactttg cggctataat ttccttatca aacatcttga tggccatcct 960
ttggttcttt ctagtaaaca aggagatgtt atcaagccag gagtcatcag aggagttctt 1020
ggaaaaggaa tgccaaataa gaaataccca gaactcaaag gaaacttgtt cgttgaattt 1080
gaagtcgaat ttccaaagga gcatttcctc gatgatgaaa aagcttatgc cgttctgaaa 1140
agctgcttcc ctacctcaaa agttgtcaat gtaaccccag ctgccgcaga agtttctctt 1200
atggaatatg acgagaagaa gtacagccga ggacgtggcg gagacgctta caatgaagat 1260
tcggacgaag aacaacacgg aggacatcac ggacaaggcg tcagatgcca acaccaatag 1320
<210>48
<211>439
<212>PRT
<213>秀丽新小杆线虫
<400>48
Met Phe Gly Gly Gly Ser Ser Gly Pro Val Asp Thr Thr Leu Tyr Thr
1 5 10 15
Thr Leu Asn Val Arg Pro Asp Ala Ser Gln Ala Asp Ile Lys Lys Ser
20 25 30
Tyr Phe Lys Leu Ala Lys Glu Tyr His Pro Asp Lys Asn Pro Asp His
35 40 45
Gly Asp Lys Phe Lys Glu Ile Ser Phe Ala Tyr Glu Val Leu Ser Ser
50 55 60
Pro Glu Lys Arg Arg Leu Tyr Asp Ala Arg Gly Leu Glu Gly Val Gln
65 70 75 80
Gly Gly Gly Ala Gly Gly Gly Gly Gly Gly Phe Pro Gly Gly Leu Phe
85 90 95
Ser His Phe Phe Gly Gly Ala Gly Gly Asp Asp Asp Asp Asp Asp Asp
100 105 110
Asp Met Gly Gly His Pro Phe Gly Gly Leu Phe Gly Gly Met Gly Gly
115 120 125
Met Gly Arg Gly Gly Pro Arg Arg Arg Lys Phe Gln Asp Thr Val His
130 135 140
Pro Leu Asn Val Thr Leu Glu Glu Leu Tyr Val Gly Lys Thr Ser Lys
145 150 155 160
Leu Lys Leu Ser Lys Lys Ala Leu Cys Lys Thr Cys Glu Gly Ser Gly
165 170 175
Gly Lys Lys Gly Glu Lys Tyr Lys Cys Asp Ala Cys Arg Gly Arg Gly
180 185 190
Val Lys Thr Ile Val Gln Gln Ile Gly Pro Gly Met Leu Gln Gln Met
195 200 205
Gln Val His Cys Asp Ala Cys Lys Gly Ser Gly Gly Lys Val Pro Ala
210 215 220
Gly Asp Lys Cys Lys Gly Cys His Gly Glu Lys Tyr Glu Asn Val Ser
225 230 235 240
Lys Ile Leu Glu Val His Val Leu Pro Gly Met Lys His Asn Asp Lys
245 250 255
Ile Thr Phe Lys Gly Asp Gly Asp Gln Ser Asp Pro Asp Gly Glu Pro
260 265 270
Gly Asp Val Val Ile Val Ile Gln Gln Lys Asp His Asp Ile Phe Lys
275 280 285
Arg Asp Gly Asp Asp Leu His Met Thr Lys Lys Leu Ser Leu Asn Glu
290 295 300
Ala Leu Cys Gly Tyr Asn Phe Leu Ile Lys His Leu Asp Gly His Pro
305 310 315 320
Leu Val Leu Ser Ser Lys Gln Gly Asp Val Ile Lys Pro Gly Val Ile
325 330 335
Arg Gly Val Leu Gly Lys Gly Met Pro Asn Lys Lys Tyr Pro Glu Leu
340 345 350
Lys Gly Asn Leu Phe Val Glu Phe Glu Val Glu Phe Pro Lys Glu His
355 360 365
Phe Leu Asp Asp Glu Lys Ala Tyr Ala Val Leu Lys Ser Cys Phe Pro
370 375 380
Thr Ser Lys Val Val Asn Val Thr Pro Ala Ala Ala Glu Val Ser Leu
385 390 395 400
Met Glu Tyr Asp Glu Lys Lys Tyr Ser Arg Gly Arg Gly Gly Asp Ala
405 410 415
Tyr Asn Glu Asp Ser Asp Glu Glu Gln His Gly Gly His His Gly Gln
420 425 430
Gly Val Arg Cys Gln His Gln
435
<210>49
<211>1194
<212>DNA
<213>人(Homo sapiens)
<400>49
atggtgaaag aaacaactta ctacgatgtt ttgggggtca aacccaatgc tactcaggaa 60
gaattgaaaa aggcttatag gaaactggcc ttgaagtacc atcctgataa gaacccaaat 120
gaaggagaga agtttaaaca gatttctcaa gcttacgaag ttctctctga tgcaaagaaa 180
agggaattat atgacaaagg aggagaacag gcaattaaag agggtggagc aggtggcggt 240
tttggctccc ccatggacat ctttgatatg ttttttggag gaggaggaag gatgcagaga 300
gaaaggagag gtaaaaatgt tgtacatcag ctctcagtaa ccctagaaga cttatataat 360
ggtgcaacaa gaaaactggc tctgcaaaag aatgtgattt gtgacaaatg tgaaggtaga 420
ggaggtaaga aaggagcagt agagtgctgt cccaattgcc gaggtactgg aatgcaaata 480
agaattcatc agataggacc tggaatggtt cagcaaattc agtctgtgtg catggagtgc 540
cagggccatg gggagcggat cagtcctaaa gatagatgta aaagctgcaa cggaaggaag 600
atagttcgag agaaaaaaat tttagaagtt catattgaca aaggcatgaa agatggccag 660
aagataacat tccatggtga aggagaccaa gaaccaggac tggagccagg cgatattatc 720
attgtgttag atcagaagga ccatgctgtt tttactcgac gaggagaaga ccttttcatg 780
tgtatggaca tacagctcgt tgaagcactg tgtggcttcc acaagccaat atctactctt 840
gacaaccgaa ccatcgtcat cacctctcat ccaggtcaga ttgtcaagca tggagatatc 900
aagtgtgtac taaatgaagg catgccaatt tatcgtagac catatgaaaa gggtcgccta 960
atcatcgaat ttaaggtaaa ctttcctgag aatggctttc tctctcctga taaactgtct 1020
ttgctggaaa aactcctacc cgagaggaag gaagtggaag agactgatga gatggaccaa 1080
gtagaactgg tggactttga tccaaatcag gaaagacggc gccactacaa tggagaagca 1140
tatgaggatg atgaacatca tcccagaggt ggtgttcagt gtcagacctc ttaa 1194
<210>50
<211>397
<212>PRT
<213>人
<400>50
Met Val Lys Glu Thr Thr Tyr Tyr Asp Val Leu Gly Val Lys Pro Asn
1 5 10 15
Ala Thr Gln Glu Glu Leu Lys Lys Ala Tyr Arg Lys Leu Ala Leu Lys
20 25 30
Tyr His Pro Asp Lys Asn Pro Asn Glu Gly Glu Lys Phe Lys Gln Ile
35 40 45
Ser Gln Ala Tyr Glu Val Leu Ser Asp Ala Lys Lys Arg Glu Leu Tyr
50 55 60
Asp Lys Gly Gly Glu Gln Ala Ile Lys Glu Gly Gly Ala Gly Gly Gly
65 70 75 80
Phe Gly Ser Pro Met Asp Ile Phe Asp Met Phe Phe Gly Gly Gly Gly
85 90 95
Arg Met Gln Arg Glu Arg Arg Gly Lys Asn Val Val His Gln Leu Ser
100 105 110
Val Thr Leu Glu Asp Leu Tyr Asn Gly Ala Thr Arg Lys Leu Ala Leu
115 120 125
Gln Lys Asn Val Ile Cys Asp Lys Cys Glu Gly Arg Gly Gly Lys Lys
130 135 140
Gly Ala Val Glu Cys Cys Pro Asn Cys Arg Gly Thr Gly Met Gln Ile
145 150 155 160
Arg Ile His Gln Ile Gly Pro Gly Met Val Gln Gln Ile Gln Ser Val
165 170 175
Cys Met Glu Cys Gln Gly His Gly Glu Arg Ile Ser Pro Lys Asp Arg
180 185 190
Cys Lys Ser Cys Asn Gly Arg Lys Ile Val Arg Glu Lys Lys Ile Leu
195 200 205
Glu Val His Ile Asp Lys Gly Met Lys Asp Gly Gln Lys Ile Thr Phe
210 215 220
His Gly Glu Gly Asp Gln Glu Pro Gly Leu Glu Pro Gly Asp Ile Ile
225 230 235 240
Ile Val Leu Asp Gln Lys Asp His Ala Val Phe Thr Arg Arg Gly Glu
245 250 255
Asp Leu Phe Met Cys Met Asp Ile Gln Leu Val Glu Ala Leu Cys Gly
260 265 270
Phe Gln Lys Pro Ile Ser Thr Leu Asp Asn Arg Thr Ile Val Ile Thr
275 280 285
Ser His Pro Gly Gln Ile Val Lys His Gly Asp Ile Lys Cys Val Leu
290 295 300
Asn Glu Gly Met Pro Ile Tyr Arg Arg Pro Tyr Glu Lys Gly Arg Leu
305 310 315 320
Ile Ile Glu Phe Lys Val Asn Phe Pro Glu Asn Gly Phe Leu Ser Pro
325 330 335
Asp Lys Leu Ser Leu Leu Glu Lys Leu Leu Pro Glu Arg Lys Glu Val
340 345 350
Glu Glu Thr Asp Glu Met Asp Gln Val Glu Leu Val Asp Phe Asp Pro
355 360 365
Asn Gln Glu Arg Arg Arg His Tyr Asn Gly Glu Ala Tyr Glu Asp Asp
370 375 380
Glu His His Pro Arg Gly Gly Val Gln Cys Gln Thr Ser
385 390 395
<210>51
<211>1230
<212>DNA
<213>酿酒酵母(Sacharomyces cereviseae)
<400>51
atggttaaag aaactaagtt ttacgatatt ctaggtgttc cagtaactgc cactgatgtc 60
gaaattaaga aagcttatag aaaatgcgcc ttaaaatacc atccagataa gaatccaagt 120
gaggaagctg cagaaaagtt caaagaagct tcagcagcct atgaaatttt atcagatcct 180
gaaaagagag atatatatga ccaatttggt gaagatggtc taagtggtgc tggtggcgct 240
ggcggattcc caggtggtgg attcggtttt ggtgacgata tcttttccca attctttggt 300
gctggtggcg cacaaagacc aagaggtccc caaagaggta aagatatcaa gcatgaaatt 360
tctgcctcac ttgaagaatt atataagggt aggacagcta agttagccct taacaaacag 420
atcctatgta aagaatgtga aggtcgtggt ggtaagaaag gcgccgtcaa gaagtgtacc 480
agctgtaatg gtcaaggtat taaatttgta acaagacaaa tgggtccaat gatccaaaga 540
ttccaaacag agtgtgatgt ctgtcacggt actggtgata tcattgatcc taaggatcgt 600
tgtaaatctt gtaacggtaa gaaagttgaa aacgaaagga agatcctaga agtccatgtc 660
gaaccaggta tgaaagatgg tcaaagaatc gttttcaaag gtgaagctga ccaagcccca 720
gatgtcattc caggtgatgt tgtcttcata gtttctgaga gaccacacaa gagcttcaag 780
agagatggtg atgatttagt atatgaggct gaaattgatc tattgactgc tatcgctggt 840
ggtgaatttg cattggaaca tgtttctggt gattggttaa aggtcggtat tgttccaggt 900
gaagttattg ccccaggtat gcgtaaggtc atcgaaggta aaggtatgcc aattccaaaa 960
tacggtggct atggtaattt aatcatcaaa tttactatca agttcccaga aaaccatttc 1020
acatcagaag aaaacttgaa gaagttagaa gaaattttgc ctccaagaat tgtcccagcc 1080
attccaaaga aagctactgt ggacgaatgt gtactcgcag actttgaccc agccaaatac 1140
aacagaacac gggcctccag gggtggtgca aactatgatt ccgatgaaga agaacaaggt 1200
ggcgaaggtg ttcaatgtgc atctcaatga 1230
<210>52
<211>409
<212>PRT
<213>酿酒酵母
<400>52
Met Val Lys Glu Thr Lys Phe Tyr Asp Ile Leu Gly Val Pro Val Thr
1 5 10 15
Ala Thr Asp Val Glu Ile Lys Lys Ala Tyr Arg Lys Cys Ala Leu Lys
20 25 30
Tyr His Pro Asp Lys Asn Pro Ser Glu Glu Ala Ala Glu Lys Phe Lys
35 40 45
Glu Ala Ser Ala Ala Tyr Glu Ile Leu Ser Asp Pro Glu Lys Arg Asp
50 55 60
Ile Tyr Asp Gln Phe Gly Glu Asp Gly Leu Ser Gly Ala Gly Gly Ala
65 70 75 80
Gly Gly Phe Pro Gly Gly Gly Phe Gly Phe Gly Asp Asp Ile Phe Ser
85 90 95
Gln Phe Phe Gly Ala Gly Gly Ala Gln Arg Pro Arg Gly Pro Gln Arg
100 105 110
Gly Lys Asp Ile Lys His Glu Ile Ser Ala Ser Leu Glu Glu Leu Tyr
115 120 125
Lys Gly Arg Thr Ala Lys Leu Ala Leu Asn Lys Gln Ile Leu Cys Lys
130 135 140
Glu Cys Glu Gly Arg Gly Gly Lys Lys Gly Ala Val Lys Lys Cys Thr
145 150 155 160
Ser Cys Asn Gly Gln Gly Ile Lys Phe Val Thr Arg Gln Met Gly Pro
165 170 175
Met Ile Gln Arg Phe Gln Thr Glu Cys Asp Val Cys His Gly Thr Gly
180 185 190
Asp Ile Ile Asp Pro Lys Asp Arg Cys Lys Ser Cys Asn Gly Lys Lys
195 200 205
Val Glu Asn Glu Arg Lys Ile Leu Glu Val His Val Glu Pro Gly Met
210 215 220
Lys Asp Gly Gln Arg Ile Val Phe Lys Gly Glu Ala Asp Gln Ala Pro
225 230 235 240
Asp Val Ile Pro Gly Asp Val Val Phe Ile Val Ser Glu Arg Pro His
245 250 255
Lys Ser Phe Lys Arg Asp Gly Asp Asp Leu Val Tyr Glu Ala Glu Ile
260 265 270
Asp Leu Leu Thr Ala Ile Ala Gly Gly Glu Phe Ala Leu Glu His Val
275 280 285
Ser Gly Asp Trp Leu Lys Val Gly Ile Val Pro Gly Glu Val Ile Ala
290 295 300
Pro Gly Met Arg Lys Val Ile Glu Gly Lys Gly Met Pro Ile Pro Lys
305 310 315 320
Tyr Gly Gly Tyr Gly Asn Leu Ile Ile Lys Phe Thr Ile Lys Phe Pro
325 330 335
Glu Asn His Phe Thr Ser Glu Glu Asn Leu Lys Lys Leu Glu Glu Ile
340 345 350
Leu Pro Pro Arg Ile Val Pro Ala Ile Pro Lys Lys Ala Thr Val Asp
355 360 365
Glu Cys Val Leu Ala Asp Phe Asp Pro Ala Lys Tyr Asn Arg Thr Arg
370 375 380
Al Arg Gly Gly Ala Asn Tyr Asp Ser Asp Glu Glu Glu Gln Gly
385 390 395 400
Gly Glu Gly Val Gln Cys Ala Ser Gln
405
<210>53
<211>1239
<212>DNA
<213>人
<400>53
atggctaacg tggctgacac gaagctgtac gacatcctgg gcgtcccgcc cggcgccagc 60
gagaacgagc tgaagaaggc atacagaaag ttagccaagg aatatcatcc tgataagaat 120
ccaaatgcag gagacaaatt taaagaaata agttttgcat atgaagtact atcaaatcct 180
gagaagcgtg agttatatga cagatacgga gagcaaggtc ttcgggaagg cagcggcgga 240
ggtggtggca tggatgatat tttctctcac atttttggtg ggggattgtt cggcttcatg 3G0
ggcaatcaga gtagaagtcg aaatggcaga agaagaggag aggacatgat gcatccactc 360
aaagtatctt tagaagatct gtataatggc aagacaacca aactacaact tagcaagaat 420
gtgctctgta gtgcatgcag tggccaaggc ggaaagtctg gagctgtcca aaagtgtagt 480
gcttgtcgag gtcgaggtgt gcgcatcatg atcagacagc tggctccagg gatggtacaa 540
cagatgcagt ctgtgtgctc tgattgtaat ggagaaggag aggtaattaa tgaaaaagac 600
cgctgtaaaa aatgtgaagg gaagaaggtg attaaagaag tcaagattct tgaagtccac 660
gtagacaaag gcatgaaaca tggacagaga attacattca ctggggaagc agaccaggcc 720
ccaggagtgg aacccggaga cattgttctt ttgctacagg agaaagaaca tgaggtattt 780
cagagagatg ggaatgattt gcacatgaca tataaaatag gacttgttga agctctatgt 840
ggatttcagt tcacatttaa gcaccttgat ggacgtcaga ttgtggtgaa atacccccct 900
ggcaaagtaa ttgaaccagg gtgtgttcgt gtagttcgag gtgaagggat gccgcagtat 960
cgtaatccct ttgaaaaagg tgatctttac ataaagtttg atgtgcagtt tcctgaaaac 1020
aactggatca acccagacaa gctttctgaa ctagaagatc ttctgccatc tagaccggaa 1080
gttcctaaca taattggaga aacagaggag gtagagcttc aggaatttga tagcactcga 1140
ggctcaggag gtggtcagag gcgtgaagcc tataatgata gctctgatga agaaagcagc 1200
agccatcatg gacctggagt gcagtgtgcc catcagtaa 1239
<210>54
<211>412
<212>PRT
<213>人
<400>54
Met Ala Asn Val Ala Asp Thr Lys Leu Tyr Asp Ile Leu Gly Val Pro
1 5 10 15
Pro Gly Ala Ser Glu Asn Glu Leu Lys Lys Ala Tyr Arg Lys Leu Ala
20 25 30
Lys Glu Tyr His Pro Asp Lys Asn Pro Asn Ala Gly Asp Lys Phe Lys
35 40 45
Glu Ile Ser Phe Ala Tyr Glu Val Leu Ser Asn Pro Glu Lys Arg Glu
50 55 60
Leu Tyr Asp Arg Tyr Gly Glu Gln Gly Leu Arg Glu Gly Ser Gly Gly
65 70 75 80
Gly Gly Gly Met Asp Asp Ile Phe Ser His Ile Phe Gly Gly Gly Leu
85 90 95
Phe Ile Phe Met Gly Asn Gln Ser Arg Ser Arg Asn Gly Arg Arg Arg
100 105 110
Gly Glu Asp Met Met His Pro Leu Lys Val Ser Leu Glu Asp Leu Tyr
115 120 125
Asn Gly Lys Thr Thr Lys Leu Gln Leu Ser Lys Asn Val Leu Cys Ser
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Ala Cys Arg Gly Arg Gly Val Arg Ile Met Ile Arg Gln Leu Ala Pro
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Gly Met Val Gln Gln Met Gln Ser Val Cys Ser Asp Cys Asn Gly Glu
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Lys Val Ile Lys Glu Val Lys Ile Leu Glu Val His Val Asp Lys Gly
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Pro Gly Val Glu Pro Gly Asp Ile Val Leu Leu Leu Gln Glu Lys Glu
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275 280 285
Leu Asp Gly Arg Gln Ile Val Val Lys Tyr Pro Pro Gly Lys Val Ile
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325 330 335
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Glu Glu Val Glu Leu Gln Glu Phe Asp Ser Thr Arg Cly Ser Gly Gly
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Gly Gln Arg Arg Glu Ala Tyr Asn Asp Ser Ser Asp Glu Glu Ser Ser
385 390 395 400
Ser His His Gly Pro Gly Val Gln Cys Ala His Gln
405 410
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gagaacgagc tgaagaaggc ataccgaaag ttagccaaag aataccaccc tgataagaat 120
ccaaatgctg gagacaaatt taaagaaata agttttgcat atgaagtatt gtcaaatcca 180
gagaagcgag agctgtatga cagatatgga gaacaaggcc tacgggaagg cagcggcgga 240
ggcggtggca tggatgatat cttctcacat atttttggtg gaggattgtt tggctttatg 300
ggcaatcaga gtagaagtcg aaatggcaga agaagaggcg aggacatgat gcatccacta 360
aaagtatctt tagaagacct gtacaatggc aagacaacca aactacaact tagcaagaat 420
gtgctctgta gtgcatgcag tggccaaggt gggaagtctg gagctgttca gaaatgcagc 480
gcttgtcggg gtcgaggtgt gcgcattatg atcagacagc tggctccagg aatggtgcag 540
cagatgcagt ccgtgtgctc cgactgtaat ggagaagggg aggtcatcaa tgaaaaagac 600
cgctgtaaaa aatgtgaagg gaagaaggta atcaaagaag tcaagattct ggaagtccat 660
gtagacaaag gcatgaaaca tggacagagg attacgttca ctggggaagc agaccaggct 720
ccaggagtgg aacctggaga tattgttctt ttgctacagg aaaaagaaca tgaggtgttc 780
cagagagatg ggaatgattt gcatatgaca tataagatag gactcgttga agctttatgt 840
ggatttcagt tcacatttaa acatcttgat gctcgtcaga ttgtggtgaa atacccccct 900
ggcaaagtaa ttgaaccagg atgtgttcgt gttgttcgag gtgaaggaat gccacagtat 960
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gactggatca acccagacaa actttctgaa ttagaagatc tcctgccatc tagaccagaa 1080
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ggctctggcg gtggtcagag acgtgaagcc tataatgata gctctgatga agaaagtagc 1200
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Gly Met Val Gln Gln Met Gln Ser Val Cys Ser Asp Cys Asn Gly Glu
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Gly Glu Val Ile Asn Glu Lys Asp Arg Cys Lys Lys Cys Glu Gly Lys
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Lys Val Ile Lys Glu Val Lys Ile Leu Glu Val His Val Asp Lys Gly
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<210>57
<211>654
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cttctacatc ggcttaggtg tagcaacacg actttattat tattattatt attattatta 60
ttattttaca aaaatataaa atagatcagt ccctcaccac aagtagagca agttggtgag 120
ttattgtaaa gttctacaaa gctaatttaa aagttattgc attaacttat ttcatattac 180
aaacaagagt gtcaatggaa caatgaaaac catatgacat actataattt tgtttttatt 240
attgaaatta tataattcaa agagaataaa tccacatagc cgtaaagttc tacatgtggt 300
gcattaccaa aatatatata gcttacaaaa catgacaagc ttagtttgaa aaattgcaat 360
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gtgcacctaa cagaatatcc aaataatatg actcacttag atcataatag agcatcaagt 540
aaaactaaca ctctaaagca accgatggga aagcatctat aaatagacaa gcacaatgaa 600
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<213>人工序列
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Claims (25)
1.相对于在非胁迫条件下生长的对应野生型植物的产量,增加在相当条件下生长的植物的植物产量的方法,其包括在植物细胞的细胞溶胶中优先增加I型DnaJ样多肽或其同源物的活性,以及任选的选择具有增加的产量的植物,其中所述I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序。
2.根据权利要求1的方法,其中所述增加的活性通过优选在编码所述I型DnaJ样多肽或其同源物的基因的基因座中引入遗传修饰实现。
3.根据权利要求2的方法,其中所述遗传修饰通过定点诱变、定向进化、同源重组、TILLING和T-DNA激活之一实现。
4.相对于在非胁迫条件下生长的对应野生型植物的产量,增加在相当条件下生长的植物的植物产量的方法,其包括在植物细胞的细胞溶胶中引入和/或表达外源I型DnaJ样核酸或其变体,所述核酸编码I型DnaJ样多肽或其同源物,所述多肽或其同源物在其羧基末端包含CaaX基序。
5.根据权利要求4的方法,其中所述I型DnaJ样核酸或其变体是原核或真核来源的,优选是真核来源的,更优选所述I型DnaJ样核酸或其变体是植物来源的,如单子叶植物,所述核酸优选来自禾本科,更优选来自稻。
6.根据权利要求4或5的方法,其中所述I型DnaJ样核酸或其变体与种子特异性启动子有效连接。
7.根据权利要求6的方法,其中所述种子特异性启动子是胚乳特异性启动子。
8.根据权利要求7的方法,其中所述胚乳特异性启动子是稻谷醇溶蛋白RP6启动子。
9.根据权利要求1到8中任一项的方法,其中所述增加的植物产量是增加的种子产量,优选是增加的收获指数。
10.通过权利要求1到9中任一项的方法得到的植物。
11.构建体,其包含:
(i)编码I型DnaJ样多肽或其同源物的核酸或其变体,所述I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序;以及
(ii)一种或多种驱动(i)的核酸序列表达的调控序列;以及任选的
(iii)转录终止序列。
12.根据权利要求11的构建体,其中所述调控序列是种子特异性启动子。
13.根据权利要求12的构建体,其中所述种子特异性启动子是胚乳特异性启动子。
14.根据权利要求13的构建体,其中所述胚乳特异性启动子是稻谷醇溶蛋白RP6启动子。
15.用权利要求11到14中任一项的构建体转化的植物。
16.产生具有增加的产量的转基因植物的方法,所述方法包括:
(i)在植物、植物部分或植物细胞的细胞溶胶中引入和/或表达编码I型DnaJ样多肽或其同源物的核酸或其变体,所述I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序;以及
(ii)在促进植物生长和发育的非胁迫生长条件下栽培植物、植物部分或植物细胞。
17.在非胁迫生长条件下具有增加的产量的转基因植物,所述增加的产量通过在所述植物中引入和/或表达I型DnaJ样核酸或其变体产生,其中所述I型DnaJ样核酸编码I型DnaJ样多肽或其同源物,所述I型DnaJ样多肽或其同源物在其羧基末端包含CaaX基序。
18.根据权利要求10、15或17的转基因植物,其中所述植物是单子叶植物,如甘蔗,或者其中植物是谷类,如稻、玉米、小麦、大麦、小米、黑麦、燕麦或高粱。
19.根据权利要求10、15、17或18中任一项的植物的可收获部分。
20.产物,其来自,优选直接来自权利要求18的植物和/或权利要求19的植物可收获部分。
21.根据权利要求19的可收获部分,和/或根据权利要求20的来自或直接来自植物的产物,其中所述可收获部分是种子。
22.I型DnaJ样核酸/基因或其变体的用途,或者I型DnaJ样多肽或其同源物的用途,其用于在非胁迫生长条件下生长的植物中增加植物产量。
23.根据权利要求23的用途,其中所述植物产量是增加的种子产量,优选是增加的收获指数。
24.根据权利要求11到14中任一项的构建体的用途,其用于在非胁迫生长条件下生长的植物中增加植物产量。
25.I型DnaJ样核酸/基因或其变体的用途,或I型DnaJ样多肽或其同源物的用途,其用作分子标记。
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EP04106985.7 | 2004-12-24 | ||
US64168805P | 2005-01-06 | 2005-01-06 | |
US60/641,688 | 2005-01-06 | ||
PCT/EP2005/057167 WO2006067236A1 (en) | 2004-12-24 | 2005-12-23 | Plants having increased yield and method for making the same |
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CN101128590A true CN101128590A (zh) | 2008-02-20 |
CN101128590B CN101128590B (zh) | 2013-01-02 |
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CN2005800486343A Expired - Fee Related CN101128590B (zh) | 2004-12-24 | 2005-12-23 | 产量增加的植物及其制备方法 |
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US (2) | US7872173B2 (zh) |
EP (1) | EP1833971B1 (zh) |
CN (1) | CN101128590B (zh) |
AR (1) | AR051866A1 (zh) |
AT (1) | ATE528404T1 (zh) |
AU (1) | AU2005318112B2 (zh) |
BR (1) | BRPI0519260A2 (zh) |
CA (1) | CA2594554A1 (zh) |
ES (1) | ES2375488T3 (zh) |
MX (1) | MX2007007389A (zh) |
WO (1) | WO2006067236A1 (zh) |
ZA (1) | ZA200704606B (zh) |
Cited By (1)
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WO2012059849A1 (en) * | 2010-11-05 | 2012-05-10 | Basf Plant Science Company Gmbh | Method for increasing yield and fine chemical production in plants |
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AR051866A1 (es) * | 2004-12-24 | 2007-02-14 | Cropdesign Nv | Plantas con aumento del rendimiento y metodo de prepracion |
US7939711B2 (en) * | 2006-09-11 | 2011-05-10 | The Chinese University Of Hong Kong | Abiotic stress tolerance conferred by J-domain containing proteins |
WO2011088065A1 (en) * | 2010-01-12 | 2011-07-21 | Monsanto Technology Llc | Transgenic plants with enhanced agronomic traits |
WO2018234191A1 (en) * | 2017-06-19 | 2018-12-27 | University Of Copenhagen | INCREASED STRENGTH IN DROUGHT IN PLANTS |
CN109678940B (zh) * | 2017-10-18 | 2022-05-10 | 中国科学院植物研究所 | 蛋白BhDnaJ6及其编码基因与应用 |
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CA2178729A1 (en) | 1993-12-09 | 1995-06-15 | Eric B. Kmiec | Compounds and methods for site-directed mutations in eukaryotic cells |
US5922929A (en) * | 1997-05-02 | 1999-07-13 | University Of Maryland At Baltimore County | Thermotolerance enhancing protein |
US6245969B1 (en) | 1997-06-24 | 2001-06-12 | Joanne Chory | Receptor kinase, Bin1 |
US6156954A (en) | 1998-07-21 | 2000-12-05 | The Salk Institute For Biological Studies | Receptor-like protein kinase, RKN, and method of use for increasing growth and yield in plants |
CN1279172C (zh) | 1999-07-22 | 2006-10-11 | 独立行政法人农业生物资源研究所 | 转化水稻植物的方法 |
US20020199218A1 (en) | 1999-08-19 | 2002-12-26 | Daphne Goring | Proline-rich extensin-like receptor kinases |
US20030044972A1 (en) * | 2000-03-17 | 2003-03-06 | Zoran Ristic | Maize chloroplast protein synthesis elongation factors and methods of use for same |
US20010052140A1 (en) * | 2000-03-24 | 2001-12-13 | Bruce Wesley B. | Methods of selection and development of plants having improved root quality and root lodging resistance |
US20040181830A1 (en) * | 2001-05-07 | 2004-09-16 | Kovalic David K. | Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement |
US7109033B2 (en) * | 2000-08-24 | 2006-09-19 | The Scripps Research Institute | Stress-regulated genes of plants, transgenic plants containing same, and methods of use |
EP1379669A2 (en) * | 2001-02-23 | 2004-01-14 | Vlaams Interuniversitair Instituut voor Biotechnologie vzw. | Plant stress regulated genes |
US20050074763A1 (en) | 2001-03-05 | 2005-04-07 | Wangxia Wang | Denaturat stable and/or protease resistant, chaperone-like oligomeric proteins, polynucleotides encoding same and their uses |
CA2491705C (en) * | 2002-07-19 | 2014-06-17 | Ipk-Institut Fuer Pflanzengenetik Und Kulturpflanzenforschung Gmbh | Method for the production of transgenic plants with increased virus resistance by silencing vegetable dnaj-like proteins |
CN1300170C (zh) * | 2002-08-19 | 2007-02-14 | 第二军医大学免疫学研究所 | 新型DnaJ/Hsp40分子DC-DJIII,其编码序列及用途 |
AU2003299859B2 (en) | 2002-12-26 | 2008-05-22 | Syngenta Participations Ag | Stress-related polypeptides and uses therefor |
US7705201B2 (en) * | 2004-06-23 | 2010-04-27 | Monsanto Technology Llc | Transgenic plants expressing cytokinin biosynthetic genes and methods of use therefor |
AR051866A1 (es) * | 2004-12-24 | 2007-02-14 | Cropdesign Nv | Plantas con aumento del rendimiento y metodo de prepracion |
NL1033850C2 (nl) | 2007-05-15 | 2008-11-18 | 3Force B V | Brandersysteem met voorgemengde branders en vlam-overdrachtsmiddelen. |
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- 2005-12-22 AR ARP050105518A patent/AR051866A1/es not_active Application Discontinuation
- 2005-12-23 EP EP05823594A patent/EP1833971B1/en not_active Not-in-force
- 2005-12-23 AT AT05823594T patent/ATE528404T1/de not_active IP Right Cessation
- 2005-12-23 AU AU2005318112A patent/AU2005318112B2/en not_active Ceased
- 2005-12-23 CN CN2005800486343A patent/CN101128590B/zh not_active Expired - Fee Related
- 2005-12-23 MX MX2007007389A patent/MX2007007389A/es active IP Right Grant
- 2005-12-23 ES ES05823594T patent/ES2375488T3/es active Active
- 2005-12-23 WO PCT/EP2005/057167 patent/WO2006067236A1/en active Application Filing
- 2005-12-23 CA CA002594554A patent/CA2594554A1/en not_active Abandoned
- 2005-12-23 US US11/794,004 patent/US7872173B2/en not_active Expired - Fee Related
- 2005-12-23 BR BRPI0519260-9A patent/BRPI0519260A2/pt not_active IP Right Cessation
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2007
- 2007-06-22 ZA ZA200704606A patent/ZA200704606B/xx unknown
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- 2011-01-17 US US13/007,746 patent/US20110179527A1/en not_active Abandoned
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2012059849A1 (en) * | 2010-11-05 | 2012-05-10 | Basf Plant Science Company Gmbh | Method for increasing yield and fine chemical production in plants |
CN103282500A (zh) * | 2010-11-05 | 2013-09-04 | 巴斯夫植物科学有限公司 | 用于增加植物中产量和精细化学品生产的方法 |
Also Published As
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ZA200704606B (en) | 2008-11-26 |
ATE528404T1 (de) | 2011-10-15 |
BRPI0519260A2 (pt) | 2009-01-06 |
US7872173B2 (en) | 2011-01-18 |
AU2005318112A1 (en) | 2006-06-29 |
EP1833971A1 (en) | 2007-09-19 |
ES2375488T3 (es) | 2012-03-01 |
US20110179527A1 (en) | 2011-07-21 |
AU2005318112B2 (en) | 2011-02-24 |
WO2006067236A1 (en) | 2006-06-29 |
MX2007007389A (es) | 2007-09-04 |
AR051866A1 (es) | 2007-02-14 |
US20090126039A1 (en) | 2009-05-14 |
CA2594554A1 (en) | 2006-06-29 |
CN101128590B (zh) | 2013-01-02 |
EP1833971B1 (en) | 2011-10-12 |
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