与玉米籽粒发育相关的bZIP转录因子、其编码基因及其基因表达的制作方法

专利名称：与玉米籽粒发育相关的bZIP转录因子、其编码基因及其基因表达的制作方法
技术领域：
本发明涉及与玉米籽粒发育相关的13个新的bZIP转录因子及其编码基因，并提 供了这些转录因子基因在籽粒发育过程中的表达特征。
背景技术：
玉米(&"ma"L),又称苞米，是世界上第三大粮食作物，播种面积仅次于水稻 和小麦。从2000年起，玉米的总产量和单产量均居第一位。玉米不仅是高产的粮食 作物，也是营养丰富的词料作物和良好的工业原料作物。玉米籽粒是玉米最主要的生 产性状。除了能为人们提供粮食来源外，实际上也是重要的繁殖器官，是未本科种子 发育研究的重要模型。玉米籽粒的发育是从受精开始的，通过双受精过程，从花粉管 尖端释放出的一个精子与卵细胞融合，形成二倍体的合子，发育成胚；另一个精子与 两个极核融合后发育成三倍体的胚乳。在整个发育过程中，籽粒经过早期胚胎发生、 成熟以及休眠，然后在适当的温度和水份条件下，开始萌发。
从分子生物学的角度来说，籽粒的发育和萌发是一个有次序的、选择性的基因表 达过程,而转录因子在这一过程中起到了关键性的作用。转录因子(transcription factor) 也称为反式作用因子(trans-actingfactor),是一类能够与真核基因启动子区顺式作用元 件(cis-acting factor)特异性结合的DNA结合蛋白。通过转录因子与顺式作用元件之间 以及它们和其他相关蛋白之间的相互作用，转录因子能够激活或抑制下游基因的转 录。根据转录因子DNA结合域的保守性，典型的植物转录因子常包括bZIP结构域、 锌指(Zinc-Finger)结构域、MADS结构域、MYC结构域、MYB结构域、Homeo 结构域、bHLH (碱性螺旋-环-螺旋)结构域及AP2/EREBP结构域等。
碱性亮氨酸拉链(basic leucine zipper，bZIP)蛋白是真核生物的转录因子中最大 且最保守的一类转录因子，该类转录因子的共同特点是(1)含有与特异DNA序列 相结合的碱性结构域；(2)参与寡聚化作用的亮氨酸拉链区与碱性区紧密相连；(3) 转录因子的N-末端含有酸性激活区；(4)以二聚体的形式结合DNA，肽链N-末端的碱 性区与DNA直接结合。在高等植物中，bZIP蛋白与种子贮藏基因表达、光形态发生 及器官建成的控制相关。
在玉米籽粒中，关于bZIP类转录因子的研究相对较少。作为玉米籽粒中研究的最 透彻的一种编码bZIP转录因子的基因，Opaque2((92)基因不仅调控玉米重要的贮藏蛋 白，如a-醇溶蛋白的特异性表达，同时还调节玉米6-马cy尸尸Z)A7基因的表达。此外， 玉米中其它的bZIP转录因子也有少量研究，如05F3./和05F3.2与小麦的HBP-lb同 源，在种子萌发的初期表达，调控细胞周期相关的组氨酸基因(Rhonda等，1993; Tetsuya等，1991);与小麦EmBP-l同源的mEmBP-l虽然在胚乳中表达量很低，但仍 能识别22kDaa-醇溶蛋白启动子区域的02-6ox(Leslie等，2004);在种子脱水过程中， v4S/能调节ABA应答基因的表达(Niu等，2002)。这些研究都表明，bZIP转录因子 对高等植物基因表达与调控十分重要。
然而，在玉米中，特别是玉米籽粒中，仍有大量的bZIP转录因子还没有被发现 和深入研究。新的转录因子的发现和研究，将对基因工程改造玉米籽粒性状提供重要 帮助。随着模式植物拟南芥和水稻基因组的测序完成，人们对转录因子的研究已经从 单个基因层面转移到了基因组层面。Riechmann(2000)的研究表明在拟南芥基因组中 约有6%的基因是编码转录因子的，Duan(2005)的研究也显示在水稻种子中特异表达 的转录因子有135个。为解析玉米籽粒发育过程中的基因表达调控，以及这些转录因 子的基因工程应用提供了基础。

发明内容
本发明的目的之一在于提供与玉米籽粒发育相关的13个新的bZIP类转录因子，
命名为MSBZIP (Maize Seed bZIP) 1-13。
本发明的目的之二在于提供该转录因子的编码基因。
本发明的目的之三在于提供该转录因子在玉米籽粒发育过程中的表达。
为达到上述目的，本发明借助基因组学研究思路，采用了玉米籽粒全发育阶段的
转录组学分析，系统发掘了在籽粒发育过程中特异表达的转录因子，总共提供了 13
个新的bZIP转录因子基因，并对它们的表达特征进行了分析。
本发明采用如下的技术方案
与玉米籽粒发育相关的bZIP转录因子MSBZIP1-13，其特征在于该转录因子是 具有序列表中序列14-26的氨基酸残基序列的蛋白质，或者是将序列14-26的氨基 酸残基序列经过一个或几个氨基酸残基的取代、缺失或添加且具有与序列14-26 的氨基酸残基序列相同活性的由序列14-26衍生的蛋白质。
上述的转录因子是具有序列表中序列14-26的氨基酸残基序列的蛋白质。
上述的与玉米籽粒发育相关的bZIP转录因子MSBZIP1-13的编码基因，其特 征在于该编码基因具有下列核苷酸序列之一
1) 序列表中序列1-13的DNA序列；
2) 与序列表中序列1-13限定的核酸序列具有90%以上的同源性，且编码相同 功能蛋白质的DNA序列。
上述的编码基因具有序列表中序列1-13的DNA序列。
上述的与玉米籽粒发育相关的bZIP转录因子MSBZIP1-13的表达，其序列特征 在于该转录因子中MSBZIP2、MSBZIP4、MSBZIP6、MSBZIP7、MSBZIP8、MSBZIP10、 MSBZIPll、 MSBZIP12以及MSBZIP13在籽粒发育过程中持续表达的；而 MSBZIP1、 MSBZIP3、 MSBZIP5、 MSBZIP9则是在发育过程中呈阶段特异性表达。
本发明提供了 13个编码在玉米籽粒发育过程中表达的bZIP转录因子新基因，并 对它们进行了进一步的分析，按所含保守区段的差异可将这13个bZIP蛋白分为三大 类，它们均含有保守的碱性亮氨酸区段，但在进化上呈现两种不同的模式，参见图2、 图3。半定量PCR分析，参见图4，表明大多数编码bZIP转录因子的基因在籽粒 发育过程中是持续表达的，在表达上呈现阶段特异性的仅4个(约占1/3)，推测特异 的表达区间可能与玉米籽粒发育的发育进程相一致，这为今后进一步研究玉米籽粒的 表达调控网络奠定了基础。


图1显示了对13个新基因所作的blastx和pfam分析。其中两者的参数都设为E value〈l(T10。
图2显示了对bZIP转录因子的保守区段的分析。其中红色方格代表保守的碱性 亮氨酸区域，黄色和绿色分别代表保守的未知功能区段。
图3显示了运用Megabase分析软件对bZIP转录因子作的聚类分析。其中02 作为聚类分析的根。
图4显示了对bZIP转录因子在整个发育阶段的表达谱分析。泳道1-14:分别是 授粉后3、 5、 7、 9、 11、 13、 15、 18、 21、 24、 27、 30、 33、 36天的不同bZIP转录 因子的RT-PCR; cGAPDH作为内参。
具体实施例方式
下面结合具体实施例，进一步阐述本发明。应理解，这些实例仅用于说明本发明 而不用于限制本发明的范围。下列实施例中未注明具体实验条件的实验方法，通常按 照常规条件，如分子克隆(Molecular Cloning:A Laboratory Manual,3rded.)或植物分 子生物学一实验手册(Plant Molecular Biology—A Laboratory Manual, Melody S. Clark 编，Springer-verlag Berlin Heidelberg, 1997)中所述条件，或按照制造厂商所建议的 条件。
实施例一玉米籽粒发育过程中表达的bZIP转录因子新基因的获得
1. 玉米籽粒发育阶段均一化cDNA文库的构建
为了得到整个籽粒发育阶段的表达序列，收集授粉后3天-36天的玉米籽粒，将 它们等体积混合后，提取总RNA (Trizol kit, Qiagen),以经Promega公司的PolyA TtractR mRNA Isolation System III Kit纯化所得的mRNA为模板，利用基于复性动力 学的原理的cDNA均一化方案，即通过变性一复性操作，使cDNA的丰度趋于一致， 最终建立均一化的cDNA文库。
2. 编码bZIP转录因子基因的获得
采用测序通用引物T3/T7 ,双脱氧法(DYEnamic ET-Terminator Cycle Sequencing), DNA序列分析仪(MegaBACE 4500)对随机挑选的10,000个克隆进行 测序。所得序列经PhredPhrap程序组分析，得到代表籽粒发育阶段表达的EST文库。 所得序歹U经 blastx( http:〃www.ncbi.nlm.nih.gov/dbEST)禾卩 pfam(http:〃www. pfam.jouy.inra.fr)分析，共得到13个编码bZIP转录因子的新基因，参见图1，其中除 4个与水稻中已知功能bZIP转录因子同源外，其余都是9个均为功能未知的新的bZIP 蛋白，我们将其命名为MSBZIP (Maize Seed bZIP) 1-13。。
实施例二 bZIP转录因子的鉴定
运用MEME分析软件对这13个bZIP转录因子进行保守区段分析，参见如图2， 它们都含有一个保守的碱性亮氨酸区域，而MSBZIP7-12除了含有bZIP蛋白特有的 碱性区段外，还具有一个功能未知的保守区段。根据所含保守区段的个数和种类，我 们将这13个转录因子分为3类，其中MSBZIP1-6属于第一类；MSBZIP7-11归为第 二类；而第三类则包括MSBZIP12和MSBZIP13。
利用Megabase3.0软件对这13个bZIP转录因子进行聚类分析，参见图3，结果 显示MSBZIP5-11在进化距离上比较接近，而MSBZIPl-4、 MSBZIP12、 MSBZIP13 以及6>2在距离上较接近，这可能揭示了存在于这些bZIP转录因子中的两种进化模 式。
实施例三bZIP转录因子的表达谱分析
以授粉后3-36天的玉米籽粒RNA为模板，Oligo (dT) 16作引物进行反转录， 得到整个籽粒发育阶段各个时间点的cDNA。根据这13个转录因子的序列分别设计 引物，参见表l，以各个时间点的cDNA为模板进行半定量PCR， PCR产物经1X琼 脂糖凝胶电泳分析，参见图4，证明MSBZIP2、 MSBZIP4、 MSBZIP6、 MSBZIP7、 MSBZIP8、 MSBZIPIO、 MSBZIPll、 MSBZIP12以及MSBZIP13都是在籽粒发育过 程中持续表达，参见图4-A;而MSBZIP1、 MSBZIP3、 MSBZIP5、 MSBZIP9则是在 发育过程中特异性表达，参见图4-B。
表l.bZIP转录因子的引物表
基因名称 序列 一
MSBZIP1 F:5'-CAGGCATTACAGTCAGAAGGG-3，
R:5'-CGGGGGTAGTAACAGATTAGAAC-3' MSBZIP2 F: 5'-AGAACGCAAGGGGTGCTCGA-3'
R: 5'-GCCGGGGACATTCTCAAGCA-3' MSBZIP3 F: 5'-CTCTTTCACCGATACAACC陽3'
R: 5'-GCTTCACCACCTGAGTTT-3' MSBZIP4 F: 5'-CAAGTCCACAGGGTCATC-3'
R: 5'-GAGCCAACTCCTCACATT-3' MSBZIP5 F:5'-ACTAGGAAACAAAAACAACACAGCA-3'
R:5'-ATGAGAGAGAACTGAAGAGACAAAA-3' MSBZIP6 F: 5'-GGACCATGACTCCAAGTG-3'
R: 5'-CGTATCCTCCTAAACCCA-3' MSBZIP7 F: 5'-TCTCCGTGATAACCAACT-3'
R: 5'-CTCTTCTCGGTGACTACAT-3' MSBZIP8 F:5'-GATTCTTTTGGACTCTGCTGG-3'
R: 5'-GCTGAGATGTTGCCTGGTAA-3' MSBZIP9 F: 5'-TTTTGATGAGCCTGTCGT-3，
R: 5'-CGTGAACATTTGCTGGAC-3， MSBZIPIO F: 5'-CTGGGAGTGAACCAACAA-3'
R: 5'-TCCCGAGTGAATAACAGC-3' MSBZIP11 F: 5'-CCATGGTCCCGGCTTCTAAT-3'
R: 5'-CCCAAGCCACAAGAAGCACC-3' MSBZIP12 F: 5'-CGAAGTCCAATAACGAGCAT-3
R: 5'-AAGGAGGAAGCAGTTCAAGG-3 MSBZIP13 F: 5'-GTTTATGGGCACCCTGTC-3'
R: 5'-GATGCTCCAAACGACCTG-3' cGAPDH F: 5 ， -GTC ACCGTCTTTGGC ATC AG-3'
R:5'-CAACAACCGAGACATCCACA-3'
序列表
<110> 上海大学
<120>与玉米籽粒发育相关的bZIP转录因子、其编码基因及其基因表达 <160> 26
<210> 1
<211> 795
<212> DNA
<213> 玉米种玉米属(Zeama"丄.)
<400〉 1
gaattcggccattacggccgggggac咖aagcgtgctaa3C8gcaMatgctc卿ggt60
ctcgtgtccgg犯gctgcagtatattgcagagcttg卿gtagagtccaggcattacagt120
c卿鄉ggtagaagtgtctgctgaaatggagttccttagccagcagaatatcatgctag180
acctggaaaacaaagccttg33gcaaagagtgg卿gtctcaactt已tta240
agcgttttcatttgagcgggaaattggtcgtctcagaac3atatatcagc300
aacagcagcaacaacaggctcctgctcttgttcgtagtaacagcagggaccttgatgtgc360
agttcgcc犯cttgtctctgaxxcca^cacgggccgggatgctctctctg420
ggcctcttcgtacttagttctttccatccttccgttcttgtgggttctatcagtggccac480
cactgtgtctgtcatctccacca^gaaaggagaagtgcacUggattgtg540
cggaaaaaggtggatctgatgtttattatttcgggatatgcacctggcgg600
tcatcctcagctcgcgtccattggtgatgctcgaccagtcggcggtgtttagggtttttg660
tttttagctagatgcttttcaagcatcattgtaccatttttcatgctagggttctaatct720
gttactacccccgttttcctg認tgtgtgcttcaagcatcttagtcatgtttggataaa780
aaaaa795
<210> 2
<211> 1454
<212> DNA
<213> 玉米种玉米属(Zeawa^Z.)
<400> 2
cggcgcctnatccccggcgctg3gg犯ttCggccattacg gccggggggc60
gccgactgcgcaaccggcaccaaccttcgccaggttttcgtgggctcttt caccgataca120
accatcctccagaacgcaaggggtgctcgattgactctagttacttagca gaaccaaaag180
tgg卿gccagaggctattttctctccatgtctcggtcac cccacctccc240
accccgctgcccacctctgggtcctcagatcactcgaagggatgacggca tgttcacgca300
^gctgc鄉tttccatctggggatacgttcattggtgaa ccaccatgct ggctgga/tga360
tctccttgaagattctgggaagctcccaagctccctcctctcagaagggc ctgcagcgat420
tctgatgccatcttagatgcgctgtcgacattccagagcccaatttatcc aatcgaggaa480
tgtgatctaaactcaggtggtg卿ctgaggacttgttgggtgg￡LgtCg￡l 8Lggtg￡Ltg￡L￡l540
agtggttctgtggtcgaggctggctgtttctatggtccga attcgccaag acagaagagt600
cttcagagagttccatggtaaatgctgtgcttgagaatgt ccccggcaac660
cccttgcagtacctgaccatcgatgcctgcagtgatgtga atggtaatgt ggccaatgteL720
actgctgatgcctgtgatgcttttggccatcctgatcaag acaagtcatt taaaaggcgc780
tcagggcaaag3tCt兆ggtccg犯aacttcaatacatcgctgaacttga aagaactgtc840
gaatcgcttc3g￡L￡LC3ttggagctgatttggccgtcagaatgtcatcact tttccatctt900
cacaatgctc犯3c^gcaaatatttccag tcttcagcaa960
gcs犯actgatt犯gga/tggtcagacgcaggcccta已aga atgaagctga aaggttgaag1020
c已gatgtctgcccgtcaccgccggagc卿agcgtcacttcctgttatga cctgagctca1080
ttcggagcagatgctacggcagtcaattggcagatgtctgacatggcaag gctcagcctg1140
aatggaagctcctctgtccccaagaagcggctatggtatatgacagcagc cggccctaac1200
ctccctcagaagataagtcgaggcattaccCt3g3t^Cttttcgctaac cttacccagg1260
catstatttgagggcccatctttgactaggalatcagttg gattcgctga attttgcaga1320
ggctatcagcttaggcccggtttggttggggttttaggctgtttttgctt tccaaaagcc1380
aaa^gatcaacc已犯agacttttttcagaagcagttgcttttcgtatttt ttttaaacaa1440
1454
<210> 3
<211> 1036
<212> DNA
<213> 玉米种玉米属(Zeawfl"L)
<400> 3
cattacggccggggg鄉cagttggtggtacgggaacgtccgcacgccgaacccagtcat60
gtcgcaggcggcgtccatatccgctagccccgggctaacc￡LCCtC￡LgCC￡latcatgctct120
tg犯agcgagtcagactccgacagcg犯tcactgtatgaggt卿gggagttccatacga180
gcgaggt犯cagatccattg卿cgaagcg犯t犯ga^ggatggtgtccaacagggagtc240
tgcgcggcggtctaggaggagg犯ac鄉cacagttgtctgatcttgagtcacaggttga300
acgactcELaeiggtgaa犯cgcaacactgttcctgcaactttcagatgccaa^ccaacEigtt360
cagtactgcagtcac3gacaacagaatcctcaaatccgatgt卿agcgt420
ggt犯卿tggc卿ggatatggtagcgagaagtgctgtatcgtgtggcctaggcgacct480
tggcctggcaccatacgtgaactcaaggaagatgtgccaagctttgaatgtgctxacagg540
gttggatttactagggagtgatgcgttcaggggtCC33CCgcaggtacacga^gtax^gaa600
ctcaccagtac已gagcactgcaagcctagagagtctggataaccgaaagtccaacgaggt660
gaccagttgcgcggcggacatttggccctgagcttc犯gccattgatctg￡LttcaB￡LgCt720
tgctccacccgaccagagtttgtcaactcaaatagctggtggcttgagca780
gagcttctggcacaagttcgcttgaaccatatttccacat犯gctttataggatgcatca840
ctcttttgatgtcacttagtttcaagtataaaatgaagctgtctttgtaca^gctgc犯900
atgagaagtaaatgaaacgtcctggacaaacttgtg犯gtcataggatct960
atttccaggtcagatgagacagtt已tgtttacaacaaaaagccatacatgttgtttagga1020
aaaaaa1036
<210> 4 <211> 1532 <212> DNA <213> 玉米种玉米属(Ze"/w";^丄.)
<400> 4
cattacggccgggggac犯tggactagttgctgcactgcttttgtgagtcaccatgggaa60
gcagtggagctct3agccaa3caaggc3tccgccccacagg已gcagcaac120
aacctgctacctcaggtgctgC犯C3CCggctgtttatccagactggtccagcttccagg180
catatcctcc33tCCC3CC3catgggttcttcccctcacctgtggCatCa ELgtCCaCagg240
gtcatccttacatgtggggagctcagcctatgattccaccatatggagca ccgccatatg300
tcatgtaccctcctggagtgtatgcacacccatctatggcctcgggtgcacatccattta360
ctccttatgccattacttctccaaatggcaatgctgatgctactggaactactgctgtag420
ctcgtgatactgatggcaagccctctgaaggc犯agataa aagtccaeLct480
aaggaactttgggcagcttg犯C8LtgCtt3ctgg犯agaa ccccagtgaa catggta3ga540
cctcaggtgca<tc￡Lgctaatggagccacttctc己aagtggcga卿tggcagtgacagtt600
c鄉cgaaggg3gtgaagg3aattctcata3tgattc3t3 ca3gc3t3gtggac3ggaac660
cgtacgaagttcccag犯tggtgcatcacgttctccatctgagggaa犯t720
tt犯ccaagctatggcaatcatgcccatgcC3tC肌gtggcccagtaactggtccaacta780
caaacctgaatattgggatggactattgggccaacac3gcaagctctgctcctgtaattc840
atggc肌已gtacagttccaggggctgtggtcccagcagagcaatggatac900
aggatg咖gg犯犯C3gtCt犯tagggagtctgcacgca960
gatctaggttgcgtaaacaggctg组gtgaggagttggctcaacgtgctgatgttttaa1020
agcagg^aatgcttcactt观卿tga^gt33accgtatt3tg3ggaac1080
ttgt3tcei3agaa/t已attcaCt￡L￡l￡Lgg￡LaBaacttgaaggcaaacaacacaaaactgatg1140
t犯caac犯gctgcaacattctggcaatga catccagaaa a犯ggaaact1200
aatcgagctatcatgcaatgcttgcatagcatggccctaa ctagattattgggaggcgca1260
attgtagtatgcgtattaactccttactttattgcagtta attgttacca ggttgtgata1320
atgttgataccaaccttgtgtgtgggtgtttcctgtttca gatctatcttggcttatctt1380
ttagaagtcaagatgtgactga卿tgtaggctgtgacccagttcactgt3tctg8lEiat31440
gtggataagctcaaatttacaatttgtattgctgactaatattgttatgtatacctggca1500
aaaggttttctttgaaaac已331532
<210> 5
<211> 906
<212> DNA
<213> 玉米种玉米属(Zeawa^丄.)
<400> 5
aaatatggatgccttcaacacgctgcagttgCtgtC33g3agctcaatcagcagctctac60
aacagctgtaccattcgctaacccccacggttcgaacaagggcctgggggttctacatag120
ttgttactacttctgtagaaCt￡Lgg￡^￡lCa<aaaacaacacagcacsgc￡Lg180
ccgagctgagggctgaggcacctggaagaagaatctgtagccgtcgaccatgtcaaatcc240
tatgatacaactttacccctcggacaaa犯tctaataccctacatcaacacctgctcagt300
cgtcctatcagtatcaggttaccaccaaattatatcagcagtcagaaagtc已gg卿3gt360
ttcagcctgtt已caaatttccattcgatttctcgtcctcaagcgctttaagcttctcatg420
tagagatgtattttctgaggtcaagccatcacacttctcctgaagttgtttcaattcttc■
ctattttcctgctttagcaa3tctgcacgattggctacttcctcccattc540
6LgtCtgtttgcgcaatcttgagcgtcgagctgactccctatttgattgcttacgtttttg600
tctcttcagttctctctcatcctgtatagccggatcactg￡Lg^caata<tcacgtgaatt660
tgatg犯gg3gctgatattggtgctgcaataactttaxxatgcatgggtacagaagc已gt720
3ccccagt犯tccactcccatattcatagtagttgctgggCC3gCC3tggg卿ggga^c780
aggccaaaatggcatcatagggttaagcacaacaggcccaga卿tggttgtgtgggctc840
犯CtgC3g33cttcgagtaccttactccccccggccgtnatggccgataa900
906
<210> 6 <211> 863 <212> DNA <213> 玉米种玉米属(Zea附a"丄.)
<400> 6
gccagaaattg3g^a^catcttcccgtgaagcgcctaaagtcccacttcctgatcagac60
aagaaatgcccctctcagaaagctgcctagttctggcagt3taaat3gtctag犯gaEi8ig120
ccatttccttcaggcttcgttggatttatctgacaatgcttcatt已gagaggcgaatgtc180
ttcagaa已gtaat已tgtc已tattatctgaggaccatgactccaagtgcttttgagtcagc240
acttcgtcaa鄉gatgg3gagcttgcttcttacatgtctcgcttggcctcattggaatc300
tattcggaattcactggcagaggagttggtgaaactgaca g犯c犯tgtgaaaagttscg360
g已ctg犯gccgctgctgtgcctggcttaagggctgaattggaagctctgaaacaaaggcs420cttccaagctctagagtt犯tggggg咖gtgatga卿gttggaggagc480
tattgttgacct犯aagagatgteLC3gggagcaggtggatcttcttgttagccggctcca540
ggcattgggtgctcgtgtatagtgccggtatggatttgtcgtgccatcgg600
acaggtctg已gacttgagcatgttttccttttttattttgtaaccgtacaacggcactga660
aaaterttcgtttgtacacatcaa&aacaccttcggcgttgggtttaggaggatacgttgt720
tgtatactagatcaagtagtttcacagttttacgtgcattcttttttatctgtgttcatt780
gaa3tgt3tc犯atttgaalccatttgcatgtgtacctcag8Lgg^tgC3atcaatcaat840
atatnggagctctttaaaaa863
<210> 7 <211> 1257 <212> DNA <213> 玉米种玉米属(Ze"w"j^Z.)
<400> 7
cattacggccgggg3caa犯gaaacacaatgctgccattgaggttcttga60
gctgcggt犯cccacttacaatagttgataggattcaact120
gtg"tcagaaattcagtatctccgtgataaccaactgtaccc犯gactattggatcttact180
g已ccgg已tggc犯aaatgtgggaggacatgaag3tac3tcgctg已犯att240
gtcctaaatctgaagtctgttgatattcctgattccaatatgattatcac300
C3C3gCCaC3cacggcagctgcgtgatattgtag已taagtggatcaca犯ttttactgac360
gtC￡l￡l￡l￡LggeLatacataaatgctctctatggctggttgaagttgaacctt420
atacccatagagagcagcctgtagcatcgccacctaggatgcagcaacct480
cctatcaa已gcctttctttatgcctggcatgagcacctagccaagctgcctgatgatctt540
gccaagacagccattgtcacttttcgagcagtcttgg已taccgitattgggtgttcaagat600
g鄉agttga犯cag犯ggag3tCtgtgSLgcagactcgtaggg￡L￡lt￡LC￡LtgCgt￡L￡LggCC660
cgggcttttgaggattggtgccacaagcatccacagccc3gagcgatcgacgtcgatcca720
g3t鄉ggtg鄉ggacaggax^t犯gg3tgtagtcaccgagaagagattcgcagtggag780
agcttgaagagcaagcttgatga/tga犯ttgaatcccst3at犯gctatcaaagcaggtc840
ccctctcgatcctg朋ggcacatctcccagagctgttccgtgctctggicc■
13
geicttctccgacgcctccttCg￡L￡L￡ltgt3Ctcgaagctgaggctgatgtcgttgatgcag960
gatcagggaa3C犯Ctag3tggacagtatgttcagaagttaggatcagtaatatccctcc1020
cccaagtgttgt犯tgcatgtgttctacccctttctgtggtstgtgt3^鄉ctagtag1080
ctcattggstgaggattcttgtttaatttattggcttgatga犯ccatcgnggtacttgt1140
3gtgtggacatatgctggatagttagcatgtgatgaactgattacaatctgtgggggtaa1200
agccgtcgtctatgtaaatgataattttgttgagatatca1257
<210> 8 <211> 1625 <212〉 DNA <213> 玉米种玉米属(Zeflm^^丄.)
<400> 8
cattacggccggggttaaggagattgacgagtacttcttgaaa^gcagcagc犯gtggt犯60
tgacgttgtgattcttttggactctgctggtgggcggcctgatgccttag卿tggaggc120
犯gLCa犯ggCgcsggaa卿aat c caaat cagcgaaggttttctccacgctctcttggag180
ctggtcttttaaatctcagcaggctaacagtgaatccacagttctgaattctagtgcttx240
tggatatggttaccatggcaagacactagagaagctttatga卿ggeigcaaaaac t c t a300
caagttaatcaatttgcaaggctccagtattctceigtgct360
acaaaagctag犯tcaggggagcatgatagactgcatgcaga犯gagttcgggatgctat420
tg犯ga^ctgca犯ctcggataatctcattggagg犯gctgtgagtttagcatgtttttc■
catat caaatctxagagacg3ggaattatatcctcagattattgaattatctgcagggct540
tgtgcatatgtgcacgagtgccatcaggtcttgcccagca600
acttagtctcgacctgggagtgaaccaacaatgtgacattcattaccaggc660
33catctcagctggaaattgaggtgtctggatggcacactgccttctgcaatctcatcac720
gttacagcgtgcatccttaaccaatggattagtctcacagactgccttcc780
鄉tg3tggtggcttaatgaggagttcatctggaatccgtagcctctccaa^gg犯ctcc3840
gcatgctcttgacaggct已ccgg卿鄉tagceLgctg犯gc犯ta^agacctttatgtc900
aatcatacactctatagttgtacagcagagtga卿gcgccaactaaaga3g肌gtC3ga卿
taatatggatagcaagttccagacccagttagagaaacacagtgataMtcaat t caaaa1020
ctcggcacagCCtCC33ELtBttcagtatcacgaaactgga1080
tgcatttagaa卿aggtcgaggagg卿aggccagatacctaacctctgtt鄉g^aig1140
ccgagccatgac3ctg已3C3atctgcagacagcccttccaaatgtgttccatgctctgga1200
gggattctcaggagtgtgcg"tgcaggcatttg犯ggtatcagctgatgcagtg鄉Ugc1260
tgttattcactcgggagcggcttcatctgctatttgctcgtgacattgctacgatggctg1320
tgattattcattgtaagtaggcgaagtataaactggttcca>cgtgaata<cccttttcttc1380
taattttcagagttttcggaggattttatg犯gtc犯t3t3ggcat3at31440
gtcttgaatagatcac犯tgtctcaactctcaatttgaga聊gatgtg31500
aaca/tatgtggctttaaattatgg咖ggattcatgeagacttc犯ctgcacgttctcat1560
tgctagtcatetc aaaaaaattccaatgtatttgttcaga^犯aaaggg1620
aaaaa1625
<210> 9 <211> 1324 <212〉 DNA <213> 玉米种玉米属(Ze"m^^丄.)
<400> 9
cattacggccgggggacctgttgagattgatgatagatttg犯anctcat60
gctgcagttctagatag犯tgcttgcttggtgtatgacgsagtgaaggct120
ggggagcttatg犯aeiccgaaaggtgtxtcta_ttgc￡igaa180
cggggtgttaa￡LCttg￡L^Ccttgg卿agac^犯gctgctgttagccaattgcacaca240
3ggtacattgtagacatgcaatccatggattcaacagtttc已g3犯taa3tcggcttcgg300
gatagacagttattcccaaagttaatggaccttgttgatggaatggccaagatgtggagt360
tctatgtaccgccatcatagaagtcagttcctgatcatttctgg犯teiag郷tttcgag420
attcctcctattccaagggagacaactgattctcattac3tccagacttgtgagctacgt■
gacattgttagggagtggcat已cgcagtttgag￡L￡ia<ttg3tggEicaatc3ga犯gcatec540
atcagagcactcaatgcatggctgaagctcaatctcatccc犯ttgagagcaacataaag600
g卿6LggtCtcctcccctcc卿gctggtggatccacccatcaagcacctgccgcgtgcc660
tggcatgacgagctcgaaaggcttccgattgagctcgc犯犯actgccatcas犯ccttt720
gctgaagtcataagcactatcgtgcatcttc鄉a卿gg^gtaaacct犯ggaggcgg780
tgtgatgaa3ctcgtagggatctgaaccga犯gaaggcacagtttgaggactggcatcaa840
aagtacttggag￡Lgacagaccaaaac3t3gatcctgtcga900
cgcaccatcgaggaactagagattcggctg卿gaggaggtcttaggcat960
gccaggcaggtgcggg卿aatcacttgcaaacctccggacacacctgccagaacttttc1020
aggaacatgtccgattttgcttacttttgc犯agacatgtacaacaatct1080
gctgcgctgtcca犯gatgaagtct犯ggctgatgatatttgtgcgtgtgtatgtatact1140
cgacaagcagtagaatgtttgctggtaacttgtcctgatgccgtctagaatacat犯tag1200
tcggcattaattatgtttgt犯sttggaatggacattccggactccccattgtttatttc1260
catgctatgtactgcattggtgcgtgg已t3犯atctttgattgctccaaa1320
aaaa1324
<210> 10 <211> 991 <212> DNA <213〉 玉米种玉米属(Ze"ma^i:.)
<400> 10
acgaggccatccaggtccagaaccatatgcccagcaagttagtctccnng60
gcaccagacctgggagtgaaccaacaagngacactcatcgCC3gCC￡l8C￡Ltctcagctgg120
aaattgaggtgtctggatggcacactgccttctgcaatcttatcacatcgcagcgtgaat180
acgttagcatccttagccaatggattggtctcacagactgccttccggatgatggtggct240
ttcatctggaatccttagcctctctgaggaactccagcgtgctcttgaca300
ggctaccagag犯ggtagcagctgaagcgataaagacctttatgtcagtceitacactcta360
tagttgtacagca犯gtg犯gagcgcc犯cgtC3g3t肪Catggatagca420
agttccagacccagttagaga^acacagtgaaaatacaatgcaaaactcggcccagcctc480
C3aata^ig已已taactattcagcatcaaagaatggaacgaaactggatgcatttataaagc540
aggtcgaggagg卿aagcccctctgtgagggt卿ccgagccatgacac600
tgaacagtctgcagac犯gccttcgaaatgtgttccatgctctggagggattctcaggag660
tgtgcgtacaggcatttgaaggtatcsgcagatgcagtgaagttgctgttattcactcgg720 gagcagcttcaxxtgctatttcctcgtgaccttgcttcaa tgactgtgattattcattgt780
a3gtaggC333gg3taa3ctggttCC3Cgtgaatacccttcgcttctaattttcagaact840
ttgaatagctttcgggggeittatgtg犯gtC3a/tataggcataatagtct900
acaatgtctc肌cttacattttggttcttattgcatgggatgtactgctctgtacataat960
catttacattttgaccgaacc t aaaaaaaa3991
<210> 11
<211> 1355
<212> DNA
<213>玉米种玉米属(Zeflm^AyL)
<400> n
ggaettcggccattacggccg
aggagctggcgagcactcggcatggcagat
gt卿catggtcaaaggtta
gcttctaattcatctgaccggtCtg3C8g3
cgacggtttgcccaaaatcgtgaggcagca
gtgcaacagct卿gagcagt3agct3aaa
gctcgacagcagggaatcttcatttctagt
3atggagct3tgaccttcggtttgg33t3t
at犯atgagctgaggactgc已gtaaatgct
gtcgatgggattatgacgag
gctgatgtgttccatatactttcaggcatg
tggcttgggggtttxcgttcgtctgagctt
ctaactgagcagc朋ttgatggggctatcc
gatgcattgtcacaagg犯tgg鄉cattg
tcccttggtccatcangatcttcttggaac
gccatgggaaaacttggcEiccctccaaaat
cagaccttacacgtatcctg
gcaatccatgactacttttcacgtttgcgc
gacaacaaag caatttgttg tctcattxtt 60
gctagttcga ggactgaceic ctcgactgtt 120
gaaaacggac agagtggagc catggtcccg 180
tctgacaaac ctatggacca gaaggtttta 240
agaaaaagcc ggctgaggaa aaaggcatat 300
cttgcaagcc tggagcaaga actccagaaa 360
tctggagatc aaactcatac tatgagtgga 420
tctcgatggc aagaggaaca aaataagcag 480
catgcaagtg atagtgacct ccgtcttata 540
atattcaggc tgaagggcat tgctgcaaag 600
tggaaaacac ctgctgaaag gtgcttcttg 660
ctaaagctcc ttgtgaatca gcttgaacct 720
aatctgcagc朋tcttctca ncaggctgag 780
caccagtcct tggcacaaac gctggctggg 840
gtggcaaact at已tgggttc犯tggctatg 900
ttccttcccc aggctgacaa tctacgacag 960
acaatccgac aagctgctcg tgcgcttctt 1020
gcactgagtt ctctttggct tgctaggcca 1080
cgagagtaac atcacccccg ttggggtcac ctggctgcaa actcgcaaag catttgcaga ttgtattaac atcttctggt ctcatattgt
ttggatt￡L￡LC ttgt犯gtt2L tCatg￡L￡lttt
aataaaaata g犯gtacata agtgctgg犯
ttgatgaact gatctgtgaa tgtgggactg 1140
tgactggagg atcagtcctc tctgtacaag 1200
gctccagtgt tgtacaattt agggaaaagc 1260
gttattggca tattgtacaa ttgtattagc 1320
犯a犯 1355
<210> 12
<211> 978
<212> DNA
<213>玉米种玉米属(Zea附"，丄.)
<400〉 12
cgggggtcasagctcgatat
tttcagac已gtctttcagca
ggactttcagC￡Lga￡L￡t￡LtgCtgagctcaag
caactgcgtgacgctttaaatgatgcactg
scgggcg卿tgtcgaagtccaataacgag
tacagcccttcattcttccagctctcggag
cagctgccgcatcatttccaacagcctccg
C3CCCC￡l3Ctccctctcgaacatggtgcag
g織Ugggaaaggatcceitggccgtg犯g
ggcagctctatatctgcgggtgaaagcaac
ttctaagaagcctttgtttggtttagagtt
gcttcctccttccctgtagtttttttattc
ttgtgtcgatcgttgttgtcgtggtggctg
g￡Lgtttgtt3tgtgtgtggtatggttggtt
gtgattttttggcagattttcttttctcca
ttcttattctgga犯tgtgtacattgtgtt
ataacagaat tggagcggaa ggttcaaact 60
cagctgacac tattccagag agacacaact 120
ataaggttgc aggccatgga gcagcaggct 180
aggcaagaag tggagaggct t犯gatagca 240
catttcaata tgggaatgca gcacgtctcc 300
caacacacaa tgcagcagca tggaaacatt 360
ctgaatgccc cgagccacca gatgctatcc 420
ca卿atcgc ttgggcggct ccaggggctg 480
tcagaggctg aggtxgtggt gaagtc卿g 540
accaccttct agctatacaa tacgtctgcc 600
cttttgttca tagtctgaca tccttgaact 660
cattcgttac atttagttac cattgatcgg 720
attctatagg gacattggtg tgttggtttc 780
gtttcgagct agtgactgaa agggcaccga 840
cctgtttttt tgttatagaa caaccatgat 900
gctatgctaa tttgtagtaa tatttttgat %0
978
<210> 13
<211> 942
<212> DNA
<213>玉米种玉米属(Ze"wflj^丄.)
<400> 13
ggcggcgctc卿3C3gtggatccccgggcCtgElgg5LELtttggccattacggccgggggt60
aagacttgagt aat aat at c認犯gggtggtaaggc犯gcacaccattt120
ggtattcagcaaaggigctgSLgaagtccgcagatctacgaagtc犯tactg180
cacgggctgtctgatagtgatctgagtttatgggcaccct240
ggatctxacaccctgaagacttgtttcaaaatggcagtactggtttgata300
acacctcaatccagtggactg卿gggtC3gaactaaatcccagccattg360
cggagatctacatactatggatgagtatcagtctgttatc420
gggatgaaaattcatcactaaaggaa8gattatcetaeigatgga卿atcg■
atgtcccaagtgagcgtgacctttgacaga眺ggt卿g540
g已tctgcagag3C^Cttg3gtctgcaaccaggcattgatcgacatcttc600
gg犯ccgacggagg卿gcctgaggaggaagctg卿gac660
gcatcttccaccatacaggaUtgctgg3gcaactgaatgttgccaggaa720
gtttg鄉aaaacgttgctccttgcgggc已tcgtcttagttt已g已gtttg780
agggctagctgcatctacattgtgctccaggtcgtUggetgcatcccatgtacagccaag840
cagtagtgaccactgaagaacacgttatgctttatctgcagttagcagttaccctscaag900
犯3t犯Cttgcgtagttatcttttt犯g犯犯942
<210> 14
<211> 217
<212> PRT
<213>玉米种玉米属(Zeawa^L)
<400> 14
lie Arg Pro Leu Arg Pro Gly Asp Asn Lys Arg Ala Lys Gin Gin 5 10 15
Tyr Ala Gin Arg Ser Arg Val Arg Lys Leu Gin Tyr lie Ala Glu 20 25 30
Leu Glu Ser Arg Val Glu Ala Leu Gin Ser Glu Gly Val Glu Val 35 40 45
Ser Ala Glu Met Glu Phe Leu Ser Gin Gin Asn He Met Leu Asp 50 55 60
Leu Glu Asn Lys Ala Leu Lys Gin Arg Val Glu Ser Leu Ala Gin 65 70 75
Glu Gin Leu He Lys Arg Phe Gin Gin Glu Met Phe Glu Arg Glu 80 85 90
lie Gly Arg Leu Arg Thr He Tyr Gin Gin Gin Gin Gin Gin Gin 95 100 105
Ala Pro Ala Leu Val Arg Ser Asn Ser Arg Asp Leu Asp Val Gin 110 115 120
Phe Ala Gin Leu Ser Leu Lys His Lys Asp Pro Asn Thr Gly Arg 125 130 135
Asp Ala Leu Ser Gly Pro Leu Arg Thr Phe Phe Pro Ser Phe Arg 140 145 150
Ser Cys Gly Phe Tyr Gin Trp Pro Pro Leu Cys Leu Ser Ser Pro 155 160 165
Pro Arg Lys Glu Lys Cys Thr Trp lie Val Lys Thr Pro Glu Lys 170 175 180
Gly Leu Trp He Cys Leu Leu Phe Arg Asp Met His Leu Ala Val 185 l卯195
He Leu Ser Ser Arg Pro Leu Val Met Leu Asp Gin Ser Ala Val 200 205 210
Phe Arg Val Phe Val Phe Ser 215<210> 15 <211〉 420 <212> PRT
<213> 玉米种玉米属(Zeama"丄.) <400> 15
Arg Arg Leu Arg Thr Val Asp Pro Arg Arg Gly lie Arg Pro Leu 5 10 15
Arg Pro Gly Gly Ala Asp Cys Ala Thr Gly Thr Asn Leu Arg Gin 20 25 30
Val Phe Val Gly Ser Phe Thr Asn Thr Thr lie Leu Gin Asn Ala 35 40 45
Arg Gly Ala Arg Leu Thr Leu Val Thr Gin Asn Gin Lys Trp Lys 50 55 60
Ala lie Lye Asp Thr Arg Leu Phe Ser Leu His Val Ser Val Thr 65 70 75
Pro Pro Pro Thr Pro Leu Pro Thr Ser Gly Ser Ser Asp His Ser 80 85 90
Lys Gly Arg His Val His Ala Lys Leu Gin Val Ser lie Trp Gly 95 100 105
Tyr Val His Trp Thr Thr Met Leu Ala Gly Ser Pro Arg Phe Trp 110 115 120
Glu Ala Pro Lys Leu Pro Pro Leu Arg Arg Ala.Cys Ser Asp Ser 125 130 135
Asp Ser Ala lie Leu Asp Ala Leu Ser Thr Phe Gin Ser Pro lie Tyr 140 145 150
Pro He Glu Glu Cye Asp Leu Asn Ser Gly Gly Glu Ala Glu Asp 155 160 165
Leu Leu Gly Gly Val Glu Gly Asp Glu Ser Gly Ser Val Val Glu 170 175 180
Ala Gly Cys Phe Tyr Gly Pro Asn Ser Pro Arg Gin Lys Ser Arg 185 190 195
Leu Thr Thr Ser Glu Ser Ser Met Val Asn Ala Val Leu Glu Asn 200 205 210
Val Pro Gly Asn Pro Leu Gin Tyr Leu Thr lie Asp Ala Cys Ser 215 220 225
Asp Val Asn Gly Asn Val Ala Asn Val Thr Ala Asp Ala Cys Asp 230 235 240
Ala Phe Gly His Pro Asp Gin Asp Lys Ser Phe Lys Arg Arg Ser 245 250 255
Gly Asp Arg Ser Arg Val Arg Lys Leu Gin Tyr lie Ala Glu Leu 260 265 270
Glu Arg Thr Val Glu Ser Leu Gin Asn lie Gly Ala Asp Leu Ala 275 280 285
Val Arg Met Ser Ser Leu Phe His Leu His Asn Ala Leu Ser Met 290 295 300
Glu Asn Lys Gin Leu Arg He His lie Ser Ser Leu Gin Gin Ala 305 310 315
Lys Leu lie Lys Asp Gly Gin Thr Gin Ala Leu Lys Asn Glu Ala 320 325 330
Glu Arg Leu Lys Gin Met Ser Ala Arg His Arg Arg Ser Arg Ser 335 340 345
Val Thr Ser Cys Tyr Asp Leu Ser Ser Phe Gly Ala Asp Ala Thr 350 355 360
Ala Val Asn Trp Gin Met Ser Asp Met Ala Arg Leu Ser Leu Asn 365 370 375
Gly Ser Ser Ser Val Pro Lys Lys Arg Leu Trp Tyr Met Thr Ala 380 385 390
Ala Gly Pro Asn Leu Pro Gin Lys lie Ser Arg Gly lie Thr Leu 395 400 405
Asp Asn Phe Ser Leu Thr Leu Pro Rro His lie Phe Glu Gly Pro
410 415 420
<210> 16 <211> 210 <212> PRT
<213> 玉米种玉米属(Zeflwa"丄.) <400> 16
Met Ser Gin Ala Ala Ser lie Ser Ala Ser Pro Gly Leu Thr Thr 5 10 15
Ser Ala Asn His Ala Leu Glu Ser Glu Ser Asp Ser Asp Ser Glu 20 25 30
Ser Leu Tyr Glu Val Glu Gly Val Pro Tyr Glu Arg Gly Asn Arg 35 40 45
Ser lie Glu Thr Lys Arg lie Arg Arg Met Val Ser Asn Arg Glu 50 55 60
Ser Ala Arg Arg Ser Arg Arg Arg Lys Gin Ala Gin Leu Ser Asp 65 70 75
Leu Glu Ser Gin Val Glu Arg Leu Lys Gly Glu Asn Ala Thr Leu 80 85 90
Phe Leu Gin Leu Ser Asp Ala Asn Gin Gin Phe Ser Thr Ala Val 95 100 105
Thr Asp Asn Arg lie Leu Lys Ser Asp Val Glu Ala Leu Arg lie 110 115 120
Lys Val Lys Met Ala Glu Asp Met Val Ala Arg Ser Ala Val Ser 125 130 135
Cys Gly Leu Gly Asp Leu Gly Leu Ala Pro Tyr Val Asn Ser Arg 140 145 150
Lys Met Cys Gin Ala Leu Asn Val Leu Thr Gly Leu Asp Leu Leu 155 160 165 Gly Ser Asp Ala Phe Arg Gly Pro Thr Ala Gly Thr Arg Val Gin 170 175 180
Asn Ser Pro Val Gin Ser Thr Ala Ser Leu Glu Ser Leu Asp Asn 185 190 195
Arg Lys Ser Asn Glu Val Thr Ser Cys Ala Ala Asp He Trp Pro 200 205 210
<210> 17 <211> 382 <212> PRT
<213> 玉米种玉米属(Zeaway;s丄.) <400> 17
Met Gly Ser Ser Gly Ala Asp Thr Ser Ser Lys Pro Asn Lys Ala 5 10 15
Ser Ala Pro Gin Glu Gin Gin Gin Pro Ala Thr Ser Gly Ala Ala 20 25 30
Thr Pro Ala Val Tyr Pro Asp T卬Ser Ser Phe Gin Ala Tyr Pro 35 40 45
Pro lie Pro Pro His Gly Phe Phe Pro Ser Pro Val Ala Ser Ser 50 55 60
Pro Gin Gly His Pro Tyr Met Trp Gly Ala Gin Pro Met lie Pro 65 70 75
Pro Tyr Gly Ala Pro Pro Tyr Val Met Tyr Pro Pro Gly Val Tyr 80 85 90
Ala His Pro Ser Met Ala Ser Gly Ala His Pro Phe Thr Pro Tyr 95 100 105
Ala lie Thr Ser Pro Asn Gly Asn Ala Asp Ala Thr Gly Thr Thr 110 115 120
Ala Val Ala Arg Asp Thr Asp Gly Lys Pro Ser Glu Gly Lys Asp 125 130 135
Lys Ser Pro Thr Lys Arg Pro Lys Gly Thr Leu Gly Ser Leu Asn 140 145 150
Met Leu Thr Gly Lys Asn Pro Ser Glu His Gly Lys Thr Ser Gly 155 160 165
Ala Ser Ala Asn Gly Ala Thr Ser Gin Ser Gly Glu Ser Gly Ser 170 175 180
Asp Ser Ser Ser Glu Gly Ser Glu Gly Asp Ser His Asn Asp Ser 185 190 195
Tyr Lys His Ser Gly Gin Glu Gin Asp Gly Asp Val Arg Ser Ser 200 205 210
Gin Asn Gly Ala Ser Arg Ser Pro Ser Glu Gly Lys Phe Asn Gin 215 220 225
Ala Met Ala lie Met Pro Met Pro Ser Ser Gly Pro Val Thr Gly 230 235 240
Pro Thr Thr Asn Leu Asn lie Gly Met Asp Tyr Trp Ala Asp Thr 245 250 255
Ala Ser Ser Ala Pro Val lie His Gly Lys Val Thr Pro Thr Thr 260 265 270
Val Pro Gly Ala Val Val Pro Ala Glu Gin Trp lie Gin Asp Glu 275 280 285
Arg Glu Leu Lys Arg Gin Lys Arg Lys Gin Ser Asn Arg Glu Ser 290 295 300
Ala Arg Arg Ser Arg Leu Arg Lys Gin Ala Glu Cys Glu Glu Leu 305 310 315
Ala Gin Arg Ala Asp Val Leu Lys Gin Glu Asn Ala Ser Leu Arg 320 325 330
Asp Glu Val Asn Arg He Arg Lys Glu Tyr Glu Tyr Leu Val Ser 335 340 345
Lys Asn Asn Ser Leu Lys Glu Lys Leu Glu Gly Lys Gin His Lys 350 355 360 Thr Asp Glu Ala Gly Leu Asn Asn Lys Leu Gin His Ser Gly Asn 365 370 375
Asp lie Gin Lys Lys Gly Asp 380
<210> 18 <211> 163 <212> PRT
<213〉 玉米种玉米属(Zeama^丄.) <400> 18
Gly Thr Arg Ser Ser Ala Val Glu Pro Thr Gin Pro Ser Ser Gly 5 10 15
Pro Val Val Leu Asn Pro Met Met Pro Phe T卬Pro Val Pro Ser 20 25 30
Pro Met Ala Gly Pro Ala Thr Thr Met Asn Met Gly Val Asn Tyr 35 40 45
Trp Gly Thr Ala Ser Val Pro Met His Gly Lys Val He Ala Ala 50 55 60
Pro He Ser Ala Pro Ser Ser Asn Ser Arg Asp lie Val Leu Ser 65 70 75
Asp Pro Ala lie Gin Asp Glu Arg Glu Leu Lys Arg Gin Lys Arg 80 85 90
Lys Gin Ser Asn Arg Glu Ser Ala Arg Arg Ser Arg Leu Arg Lys 95 100 105
Gin Thr Glu Trp Glu Glu Val Ala Asn Arg Ala Asp Leu Leu Lys 110 115 120
Gin Glu Asn Ser Ser Leu Lys Glu Glu Leu Lys Gin Leu Gin Glu 125 130 135
Lys Cys Asp Gly Leu Thr Ser Glu Asn Thr Ser Leu His Glu Lys
140 145 150
Leu Lys Ala Leu Glu Asp Glu Lys Ser Asn Gly Asn Leu 155 160
<210> 19 <211> 247 <212> PRT
<213> 玉米种玉米属(Zeawa"丄,) <400> 19
Ala Arg Asn Glu Asn lie Phe Pro Ser Ala Ser Pro Thr Ser Ser 5 10 15
Asp Lys Lys Cys Pro Ser Gin Lys Ala Ala Phe Trp Gin Tyr Lys 20 25 30
Ser Arg Arg Lys Pro Phe Pro Ser Gly Phe Val Gly Phe lie Gin 35 40 45
Cys Phe lie Arg Glu Ala Asn Val Phe Arg Lys Tyr Val lie Leu 50 55 60
Ser Glu Asp His Asp Ser Lys Cys Phe Val Ser Thr Ser Ser Lys 65 70 75
Gly Trp Arg Ala Cys Phe Leu His Val Ser Leu Gly Leu lie Gly 80 85 90
lie Tyr Ser Glu Phe Thr Gly Arg Gly Val Gly Glu Thr Asp Arg 95 100 105
Thr Met LysVal Thr Asn Ser Arg Cys Cys Ala Trp Leu Lys Gly 110 115 120
He Gly Ser Ser Glu Thr Lys Ala Leu Pro Ser Ser Arg Val Asn 125 130 135
Gly Gly Thr Arg Val Gly Gly Ala Lys Lys Tyr Cys Pro Lys Arg 140 145 150 Asp Val Gin Gly Ala Gly Gly Ser Ser Cys Pro Ala Pro Gly lie 155 160 165
Gly Cys Ser Cys He Val Pro Val T卬He Cys Arg Ala He Gly 170 175 180
Gin Gly Lys Thr Gly Leu Arg Leu Glu His Val Phe Leu Phe Tyr 185 190 195
Phe Val Thr Val Gin Arg His Lys Tyr Ser Phe Val His lie Lys 200 205 210
Asn Thr Phe Gly Val Gly Phe Arg Arg lie Arg Cys Cys lie Leu 215 220 225
Asn Gin Val Val Ser Gin Phe Tyr Val His Ser Phe Leu Ser Val 230 235 240
Phe lie Glu Met Tyr Gin He 245
<210> 20 <211> 292 <212> PRT
<213> 玉米种玉米属(Zeama^丄.) <400> 20
Met Gin Ser ivlet Asn Ser Thr Val Ser Glu lie Gin Tyr Leu Arg 5 10 15
Asp Asn Gin Leu Tyr Pro Arg Leu Leu Asp Leu Thr Asp Arg Met 20 25 30
Ala Lys Met Trp Glu Asp Met Lys lie His His Ala Asn Gin Leu 35 40 45
Lys lie Val Leu Asn Leu Lys Ser Val Asp lie Pro Asp Ser Asn 50 55 60
lie Glu Thr Ser Asp Tyr His His Ser His Thr Arg Gin Leu Arg
65 70 75
Asp He Val Asp Lys Trp lie Thr Asn Phe Thr Asp Phe Met Asn 80 85 90
Cys Gin Lys Glu Tyr He Asn Ala Leu Tyr Gly Trp Leu Lys Leu 95 100 105
Asn Leu lie Pro lie Glu Ser Ser Leu Lys Glu Lys Val Ala Ser 110 115 120
Pro Pro Arg Met Gin Gin Pro Pro lie Lys Ala Phe Leu Tyr Ala 125 130 135
Tip His Glu His Leu Ala Lys Leu Pro Asp Asp Leu Ala Lys Thr 140 145 150
Ala lie Val Thr Phe Arg Ala Val Leu Asp Thr He Leu Gly Val 155 160 165
Gin Asp Glu Glu Leu Lys Gin Lys Glu lie Cys Glu Gin Thr Arg 170 175 180
Arg Glu Tyr Met Arg Lys Ala Arg Ala Phe Glu Asp T卬Cys His 185 190 195
Lys His Pro Gin Pro Arg Ala lie Asp Val Asp Pro Asp Ser Gly 200 205 210
Glu Gly Thr Gly His Lys Asp Val Val Thr Glu Lys Arg Phe Ala 215 220 225
Val Glu Ser Leu Lys Ser Lys Leu Asp Asp Glu He Glu Ser His 230 235 240
Asn Lys Lys Ser Lys Gin Val Arg Glu Lys Ser Leu Ser lie Leu 245 250 255
Lys Ala His Leu Pro Glu Leu Phe Arg Ala Leu Thr Asp Phe Ser 260 265 270
Asp Ala Ser Phe Glu Met Tyr Ser Lys Leu Arg Leu Met Ser Leu 275 280 285
Met Gin Asp Gin Gly Asn Asn
290
<210> 21 <211> 392 <212> PRT
<213> 玉米种玉米属(Zeawa"丄.) <400> 21
Val lie Leu Leu Asn Ser Ala Gly Gly Arg Pro Asp Ala Leu Glu 5 10 15
Met Glu Ala Asn Lys Gly Ala Gly Lys Lys Ser Lys Ser Ala Lys 20 25 30
Val Phe Ser Thr Leu Ser Trp Ser Trp Ser Phe Lys Ser Gin Gin 35 40 45
Ala Asn Ser Glu Ser Thr Val Leu Asn Ser Ser Ala Ser Gly Tyr 50 55 60
Gly Tyr His Gly Lys Thr Leu Glu Lys Leu Tyr Glu Glu Glu Glu 65 70 75
Lys Leu Tyr Lys Leu lie Lys Asp Glu Glu Phe Ala Arg Leu Gin 80 85 90
Tyr Lys Lys His Val Ser Val Leu Gin Lys Leu Glu Ser Gly Glu 95 100 105
His Asp Arg Leu His Ala Glu Arg Val Arg Asp Ala lie Glu Glu 110 115 120
Leu Gin Thr Arg He lie Ser Leu Glu Glu Ala Val Ser Leu Ala 125 130 135
Cys Phe Ser lie Ser Asn Leu Arg Asp Glu Glu Leu Tyr Pro Gin 140 145 150
He lie Glu Leu Ser Ala Gly Leu Val His Met Trp Lys Asn Met 155 160 165
His Glu Cys His Gin Val Gin Asn His lie Ala Gin Gin Leu Ser 170 175 180
Leu lie Gly Asn Arg Pro Gly Ser Glu Pro Thr Ser Asn lie His 185 l卯195
Tyr Gin Ala Thr Ser Gin Leu Glu lie Glu Val Ser Gly Trp His 200 205 210
Thr Ala Phe Cys Asn Leu lie Thr Leu Gin Arg Glu Trp lie Ser 215 220 225
lie Leu Asn Gin Trp lie Ser Leu Thr Asp Cys Leu Pro Asp Asp 230 235 240
Gly Gly Leu Met Arg Ser Ser Ser Gly lie Arg Ser Leu Ser Lys 245 250 255
Glu Leu Gin His Ala Leu Asp Arg Leu Pro Glu Lys Val Ala Ala 260 265 270
Glu Ala He Lys Thr Phe Met Ser lie He His Ser lie Val Val 275 280 285
Gin Gin Ser Glu Glu Arg Gin Leu Lys Lys Lys Ser Asp Asn Met 2卯295 300
Asp Ser Lys Phe Gin Thr Gin Leu Glu Lys His Ser Asp Asn Ser 305 310 315
lie Gin Asn Ser Ala QGln Pro Pro Asn Lys Asn His Tyr Ser Val 320 325 330
Ser Lys Asn Glu Thr Lys Leu Asp Ala Phe Arg Lys Lys Val Glu 335 340 345
Glu Glu Lys Ala Arg Tyr Leu Thr Ser Val Arg Glu Ser Arg Ala 350 355 360
Met Thr Leu Asn Asn Leu Gin Thr Ala Leu Pro Asn Val Phe His 365 370 375
Ala Leu Glu Gla Phe Ser Gly Val Cys Val Gin Ala Phe Glu Gly 380 385 390
lie Ser
<210> 22 <211> 361 <212> PRT
<213> 玉米种玉米属(ZeflWfl^丄.) <400> 22
Thr lie lie Leu Arg Leu Met lie Asp Leu Lys Ala His Ala Ala 5 10 15
Val Leu Asn Arg Met Leu Ala Trp Glu Lys Lys Leu Tyr Asp Glu 20 25 30
Val Lys Ala Gly Glu Leu Met Lys Thr Asp Tyr Gin Lys Lys Val 35 40 45
Ser Leu Leu Gin Lys Gin Lys Lys Arg Gly Val Lys Leu Glu Thr 50 55 60
Leu Glu Lys Thr Lys Ala Ala Val Ser Gin Leu His Thr Arg Tyr 65 70 75
He Val Asp Met Gin Ser Met Asp Ser Thr Val Ser Glu He Asn 80 85 90
Arg Leu Arg Asp Arg Gin Leu Phe Pro Lys Leu Met Asp Leu Val 95 100 105
Asp Gly Met Ala Lys Meyt Tip Ser Ser Met Tyr Arg His His Arg 110 115 120
Ser Gin Phe Leu lie lie Ser Gly lie Arg Gly Phe Glu He Pro 125 130 135
Pro lie Pro Arg Glu Thr Thr Asp Ser His Tyr lie Gin Thr Cys 140 145 150
Glu Leu Arg Asp lie Val Arg Glu Trp His Thr Gin Phe Glu Lys 155 160 165
Leu Mey Asp Asn Gin Lys Ala Tyr He Arg Ala Leu Asn Ala Trp 170 175 180
Leu Lys Leu Asn Leu lie Pro lie Glu Ser Asn lie Lys Glu Lys 185 190 195
Val Ser Ser Pro Pro Arg Leu Val Asp Pro Pro lie Lys His Leu 200 205 210
Pro Arg Ala Trp His Asp Glu Leu Glu Arg Leu Pro lie Glu Leu 215 220 225
Ala Lys Thr Ala lie Lys Thr Phe Ala Glu Val He Ser Thr He 230 235 240
Val His Leu Gin Glu Glu Glu Val Asn Leu Arg Arg Arg Cys Asp 245 250 255
Glu Thr Arg Arg Asp Leu Asn Arg Lys Lys Ala Gin Phe Glu Asp 260 265 270
Trp His Gin Lys Tyr Leu Glu Arg Gin Thr Ala Glu Thr Gin Asn 275 280 285
lie Asp Pro Val Glu Asp Arg Lys Arg Thr He Glu Glu Leu Glu 290 295 300
lie Arg Leu Arg Glu Glu Glu Gly His His Leu Arg His Ala Arg 305 310 315
Gin Val Arg Glu Lys Ser Leu Ala Asn Leu Arg Thr His Leu Pro 320 325 330
Glu Leu Phe Arg Asn Met Ser Asp Phe Ala Tyr Phe Cys Lys Asp 335 340 345
Met Tyr Asn Asn Leu Arg Lys Thr Ala Ala Leu Ser Lys Asp Glu
350 355 360
Val
<210> 23 <211> 206
<212> PRT
<213> 玉米种玉米属(Zeawa"丄.) <400> 23
Val Ser Gly Trp His Thr Ala Phe Cys Asn Leu lie Thr Ser Gin 5 10 15
Arg Glu Tyr Val Ser lie Leu Ser Gin Trp lie Gly Leu Thr Asp 20 25 30
Cys Leu Pro Asp Asp Gly Gly Leu Met Arg Ser Ser Ser Gly lie 35 40 45
Leu Ser Leu Ser Glu Glu Leu Gin Arg Ala Leu Asp Arg Leu Pro 50 55 60
Glu Lys Val Ala Ala Glu Ala lie Lys Thr Phe Met Ser Val lie 65 70 75
His Ser He Val Val Gin Gin Ser Glu Glu Arg Gin Leu Lys Lys 80 85 90
Lys Ser Asn Asn Met Asp Ser Lys Phe Gin Thr Gin Leu Glu Lys 95 100 105
His Ser Glu Asn Thr Met Gin Asn Ser Ala Gin Pro Pro Asn Lys 110 115 120
Asn Asn Tyr Ser Ala Ser Lys Asn Gly Thr Lys Leu Asp Ala Phe 125 130 135
lie Lys Gin Val Glu Glu Glu Lys Ala Arg Tyr Leu Thr Ser Val 140 145 150
Arg Val Ser Arg Ala Met Thr Leu Asn Ser Leu Gin Thr Ser Leu 155 160 165
Arg Asn Val Phe His Ala Leu Glu Gly Phe Ser Gly Val Cys Val 170 175 180
Gin Ala Phe Glu Gly He Ser Arg Cys Ser Glu Val Ala Val He 185 l卯195
His Ser Gly Ala Ala Ser Pro Ala He Ser Ser
200 205
<210> 24 <211> 335 <212> PRT
<213> 玉米种玉米属(Zea附a;^丄.) <400> 24
Met Ala Asp Ala Ser Ser Arg Thr Asp Thr Ser Thr Val Val Asp 5 10 15
Met Asp Asp Lys Asn Gin Arg Leu Glu Asn Gly Gin Ser Gly Ala 20 25 30
Met Val Pro Ala Ser Asn Ser Ser Asp Arg Ser Asp Arg Ser Asp 35 40 45
Lys Pro Met Asp Gin Lys Val Leu Arg Arg Phe Ala Gin Asn Arg 50 55 60
Glu Ala Ala Arg Lys Ser Arg Leu Arg Lys Lys Ala Tyr Val Gin 65 70 75
Gin Leu Glu Ser Ser Lys Leu Lys Leu Ala Ser Leu Glu Gin Glu 80 85 90
Leu Gin Lys Ala Arg Gin Gin Gly lie Phe lie Ser Ser Ser Gly 95 100 105
Asp Gin Thr His Thr Met Ser Gly Asn Gly Ala Met Thr Phe Gly 110 115 120
Leu Glu Tyr Ser Arg Trp Gin Glu Glu Gin Asn Lys Gin He Asn 125 130 135
Glu Leu Arg Thr Ala Val Asn Ala His Ala Ser Asp Ser Asp Leu 140 145 150
Arg Leu lie Val Asp Gly He Met Ala His Tyr Asp Glu lie Phe 155 160 165Arg Leu Lys Gly lie Ala Ala Lys Ala Asp Val Phe His lie Leu 170 175 180
Ser Gly Met Trp Lys Thr Pro Ala Glu Arg Cys Phe Leu Trp Leu 185 190 195
Gly Gly Phe Arg Ser Ser Glu Leu Leu Lys Leu Leu Val Asn Gin 200 205 210
Leu Glu Pro Leu Thr Glu Gin Gin Leu Met Gly Leu Ser Asn Leu 215 220 225
Gin Gin Ser Ser Gly Gin Ala Glu Asp Ala Leu Ser Gin Gly Met 230 235 240
Glu Ala Leu His Gin Ser Leu Ala Gin Thr Leu Ala Gly Ser Ler 245 250 255
Gly Pro Ser Gly Ser Ser Trp Asn Val Ala Asn Tyr Met Gly Ser 260 265 270
Met Ala Met Ala Met Gly Lys Leu Gly Thr Leu Gin Asn Phe Leu 275 280 285
Pro Gin Ala Asp Asn Leu Arg Gin Gin Thr Leu His Gin Met Gin 2卯295 300
Arg lie Leu Thr lie Arg Gin Ala Ala Arg Ala Leu Leu Ala He 305 310 315
His Asp Tyr Phe Ser Arg Leu Arg Ala Leu Ser Ser Leu Trp Leu
320 325 330
Ala Arg Pro Arg Glu
335
<210> 25 <211> 214 <212> PRT
<213> 玉米种玉米属(Zeawap丄.)
<400> 25
Met Glu Gin Gin Ala Gin Leu Arg Asp Ala Leu Asn Asp Ala Leu 5 10 15
Arg Glu Glu Val Glu Arg Leu Lys He Ala Thr Gly Glu Met Ser 20 25 30
Lys Ser Asn Asn Glu His Phe Asn Met Gly Met Gin His Val Ser 35 40 45
Tyr Ser Pro Ser Phe Phe Gin Leu Ser Glu Gin His Thr Met Gin 50 55 60
Gin His Gly Asn He Gin Leu Pro His His Phe Gin Gin Pro 65 70 75
Pro Leu Asn Ala Pro Ser His Gin Met Leu Ser His Pro Asn Ser 80 85 90
Leu Ser Asn Met Val Gin Gin Glu Ser Leu Gly Arg Leu Gin Gly 95 100 105
Leu Asp lie Gly Lys Gly Ser Met Ala Val Lys Ser Glu Ala Glu 110 115 120
Val Val Val Lys Ser Glu Gly Ser Ser He Ser Ala Gly Glu Ser 125 130 135
Asn Thr Thr Phe Leu Tyr Asn Thr Ser Ala Phe Glu Ala Phe Val 140 145 150
Trp Phe Arg Val Leu Leu Phe lie Val His Pro Thr Ala Ser Ser 155 160 165
Phe Pro Val Val Phe Leu Phe His Ser Leu His Leu Val Thr lie 170 175 180
Asp Arg Leu Cys Arg Ser Leu Leu Ser Trp Trp Leu lie Leu Gly 185 190 195
His Trp Cys Val Gly Phe Glu Phe Val Met Cys Val Val Trp Leu 200 205 210 Val Val Ser Ser
<210> 26 <211> 168 <212> PRT
<213> 玉米种玉米属(Ze詣，丄.) <400> 26
Met Gly Thr Leu Ser Arg Ser Gly Ser His Thr Leu Lys Thr Asn 5 10 15
Arg Lys Leu Phe Gin Asn Gly Ser Thr Gly Leu He Thr Pro Gin 20 25 30
Ser Ser Gly Leu He Glu Arg Glu Arg Val Arg Thr Lys Ser Gin 35 40 45
Pro Leu Asp Pro Lys Ala Thr Ser Asn Gla Asp Leu His Thr Met 50 55 60
Asp Glu Tyr Gin Ser Val lie Met Gin Leu Arg Asp Glu Asn Ser 65 70 75
Ser Leu Lys Glu Arg Leu SerLys Met Glu Glu Ser Met Ser Gin 80 85 90
Glu Leu Val Val Glu Arg Asp Lys Asn Lys Ser Leu Thr Glu Arg 95 100 105
Val Glu Asp Leu Gin Arg Gin Leu Glu Ser Ala Thr Lys Glu Gin 110 115 120
Glu Ala Leu He Asp He Phe Ser Glu Glu Arg Asn Arg Arg Asp 125 130 135
Gin Glu Glu Glu Ser Leu Arg Arg Lys Leu Lys Asp Ala Ser Ser 140 145 150
Thr lie Gin Asp Leu Leu Glu Gin Leu Asn Val Ala Arg Lys Gly 155 160 165 Arg Lys Val
权利要求
1.与玉米籽粒发育相关的bZIP转录因子MSBZIP1-13，其特征在于该转录因子是具有序列表中序列14-26的氨基酸残基序列的蛋白质，或者是将序列14-26的氨基酸残基序列经过一个或几个氨基酸残基的取代、缺失或添加且具有与序列14-26的氨基酸残基序列相同活性的由序列14-26衍生的蛋白质。
全文摘要
本发明涉及到13个在玉米籽粒发育过程中表达的bZIP转录因子新基因以及对它们的序列分析和表达分析。本发明提供的13个bZIP类转录因子基因名称为MSBZIP1-MSBZIP13，其核酸序列是下列核苷酸序列之一1)序列表中1-13的DNA序列；2)与序列表中序列1-13限定的DNA序列有90％以上同源性，且编码相同功能蛋白质的DNA序列。bZIP类转录因子基因所编码的蛋白具有序列表中序列14-26的氨基酸残基序列，或者是将序列14-26的氨基酸残基序列经过一个或几个氨基酸残基的取代、缺失或添加且具有与序列14-26的氨基酸残基序列相同活性的由序列14-26衍生的蛋白质序列。本发明所公开的这13个bZIP转录因子基因是在玉米中首次报道的，这为解析玉米籽粒发育过程中的基因表达调控，以及这些转录因子的基因工程应用提供了基础。
文档编号C07K14/415GK101096674SQ20071004119
公开日2008年1月2日 申请日期2007年5月24日 优先权日2007年5月24日
发明者冯玲娜, 宋任涛, 佳 朱, 刚 王, 许政暟 申请人:上海大学