CN110092821A - OsABCB1蛋白及其编码基因和应用 - Google Patents

OsABCB1蛋白及其编码基因和应用 Download PDF

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CN110092821A
CN110092821A CN201910463067.1A CN201910463067A CN110092821A CN 110092821 A CN110092821 A CN 110092821A CN 201910463067 A CN201910463067 A CN 201910463067A CN 110092821 A CN110092821 A CN 110092821A
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谷晓峰
张平贤
刘青
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Biotechnology Research Institute of CAAS
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Abstract

本发明公开了OsABCB1蛋白及其编码基因和应用。本发明提供的蛋白质,命名为OsABCB1蛋白,获自水稻(Oryza sativa),为序列表中序列1所示的蛋白质。编码OsABCB1蛋白的核酸分子也属于本发明的保护范围。本发明还保护一种制备转基因植物的方法,包括如下步骤:在受体植物中导入抑制所述核酸分子表达的物质,得到转基因植物;与所述受体植物相比,所述转基因植株株高降低和/或叶夹角变小。本发明可用于植物的多种性状改良,对于植物育种具有重大的应用推广价值。

Description

OsABCB1蛋白及其编码基因和应用
技术领域
本发明涉及OsABCB1蛋白及其编码基因和应用。
背景技术
水稻原产于中国和印度。是世界主要粮食作物之一。中国水稻播种面占全国粮食作物的1/4,而产量则占一半以上。栽培历史已有14000~18000年。为重要粮食作物;除食用颖果外,可制淀粉、酿酒、制醋,米糠可制糖、榨油、提取糠醛,供工业及医药用;稻秆为良好饲料及造纸原料和编织材料,谷芽和稻根可供药用。
近年来,生物技术的迅速发展大大地推动了植物育种研究手段的革新与研究水平的不断提高,植物抗病虫、抗除草剂生物技术育种已开始进入实用化阶段。利用生物技术手段将外源杀虫、抗除草剂基因导入植物基因组内,打破了植物种属甚至物种之间难以杂交的天然屏障,实现了抗虫、抗除草剂基因的转移,从而使植物迅速地、定向地获得抗虫性及机械化除草,同时又能保留原有的良好农艺性状。由于转基因玉米的每株植株都具有相当程度的抗性,因而其抗虫、抗除草剂效果比人工防治防治效果要好且稳定,还能够节省人力和物力的投入,有效的节约社会资源。其他农艺性状改良转基因植物进展及应用不如抗虫、抗除草剂性状理想,主要是由于没有优良的性状改良基因,大多农艺性状主要是有一些微效的多基因控制所致,一直没有理想的基因进行操作。
发明内容
本发明的目的是提供OsABCB1蛋白及其编码基因和应用。
本发明提供的蛋白质,命名为OsABCB1蛋白,获自水稻(Oryza sativa),为如下(a1)或(a2)或(a3)或(a4):
(a1)序列表中序列1所示的蛋白质;
(a2)在(a1)所述蛋白质的N端或/和C端连接标签得到的融合蛋白;
(a3)将(a1)经过一个或几个氨基酸残基的取代和/或缺失和/或添加得到的与植物发育相关的蛋白质;
(a4)来源于水稻且与(a1)具有98%以上同一性且与植物发育相关的蛋白质。
所述植物发育体现为株高和/或叶夹角。
标签具体如表1所示。
表1标签的序列
编码OsABCB1蛋白的核酸分子也属于本发明的保护范围。
所述核酸分子具体可为如下(b1)或(b2)或(b3)或(b4):
(b1)编码区如序列表中序列2所示的DNA分子;
(b2)序列表中序列3所示的DNA分子;
(b3)来源于水稻且与(b1)或(b2)具有95%以上同一性且编码所述蛋白质的DNA分子;
(b4)在严格条件下与(b1)或(b2)限定的核苷酸序列杂交且编码所述蛋白质的DNA分子。
上述严格条件可为用6×SSC,0.5%SDS的溶液,在65℃下杂交,然后用2×SSC,0.1%SDS和1×SSC,0.1%SDS各洗膜一次。
含有所述核酸分子的表达盒、重组载体或重组微生物均属于本发明的保护范围。
本发明还保护OsABCB1蛋白的应用,为如下(c1)或(c2):
(c1)调控植物的株高性状;
(c2)调控植物的叶夹角性状。
(c1)中,所述调控为正调控,即使植物株高增加。
(c2)中,所述调控为正调控,即使植物叶夹角变大。
本发明还保护所述核酸分子的应用,为如下(d1)或(d2):
(d1)培育株高性状改变的转基因植物;
(d2)培育叶夹角性状改变的转基因植物。
(d1)中,所述株高性状改变为株高增加。
(d2)中,所述叶夹角性状改变为叶夹角变大。
本发明还保护抑制OsABCB1蛋白的物质在植物育种的应用,所述育种的目标为如下(e1)或(e2):
(e1)培育株高降低的植物;
(e2)培育叶夹角变小的植物。
本发明还保护抑制所述核酸分子的物质的应用,为如下(f1)或(f2):
(f1)培育株高降低的转基因植物;
(f2)培育叶夹角变小的转基因植物。
本发明还保护一种制备转基因植物的方法,包括如下步骤:在受体植物中导入抑制所述核酸分子表达的物质,得到转基因植物;与所述受体植物相比,所述转基因植株株高降低和/或叶夹角变小。
抑制所述核酸分子的物质具体可为通过基因编辑抑制所述核酸分子的物质。抑制所述核酸分子的物质具体可为Cas9系统。Cas9系统中,sgRNA中的靶序列结合区如序列表的序列6所示。Cas9系统中,sgRNA的编码序列如序列表的序列5所示。抑制所述核酸分子的物质具体可为具有Cas9蛋白的编码序列和特异sgRNA的编码序列的重组质粒。特异sgRNA中的靶序列结合区如序列表的序列6所示。特异sgRNA具体为序列表的序列5所示DNA对应的RNA。Cas9蛋白的编码序列具体如序列表的序列4中第2697-6968位核苷酸所示。抑制所述核酸分子的物质具体可为重组质粒SG2027。
本发明还保护一种植物育种方法,包括如下步骤:降低目的植物中OsABCB1蛋白的含量和/或活性,从而使目的植物株高降低和/或叶夹角变小。
以上任一所述株高为成株株高。
以上任一所述植物为单子叶植物或双子叶植物。以上任一所述植物为禾本科植物。以上任一所述植物为稻属植物。以上任一所述植物为水稻,例如水稻日本晴。
本发明可用于植物的多种性状改良,对于植物育种具有重大的应用推广价值。
附图说明
图1为重组质粒SG2027的结构示意图。
图2为突变位点及其周边核苷酸的测序结果。
图3为成熟期植株的照片。
图4为成熟期植株的株高。
图5为成熟稻穗的照片。
图6为成熟稻穗的穗长。
图7为成熟籽粒的照片。
图8为千粒重的结果。
图9为粒长和粒宽的结果。
具体实施方式
以下的实施例便于更好地理解本发明,但并不限定本发明。下述实施例中的实验方法,如无特殊说明,均为常规方法。下述实施例中所用的试验材料,如无特殊说明,均为自常规生化试剂商店购买得到的。以下实施例中的定量试验,均设置三次重复实验,结果取平均值。
实施例、
OsABCB1蛋白如序列表的序列1所示。水稻日本晴的cDNA中,编码OsABCB1蛋白的CDS如序列表的序列2所示。水稻日本晴的基因组DNA中,编码OsABCB1蛋白的基因如序列表的序列3所示。
一、构建重组质粒
构建重组质粒SG2027。重组质粒SG2027的结构示意图见图1。经全质粒测序,重组质粒SG2027如序列表的序列4所示。序列表的序列4中,第2697-6968位核苷酸编码Cas9蛋白。重组质粒SG2027中,sgRNA的编码区如序列表的序列5所示(96bp)。sgRNA中的靶序列结合区如序列表的序列6所示。
二、进行遗传转化并获得再生植株
将重组质粒SG2027导入农杆菌EHA105,得到重组农杆菌。采用农杆菌浸染法,将重组农杆菌对水稻日本晴的胚性愈伤组织进行遗传转化,然后筛选抗性愈伤组织(抗性筛选采用100mg/L潮霉素),然后进行分化再生培养,然后进行生根培养,得到再生植株。
三、获得转基因植株及其后代植株
将步骤二得到的再生植株进行如下鉴定:取叶片,提取基因组DNA,采用引物GP2027-2018-F和引物GP2027-2018-R组成的引物对进行PCR扩增,然后将PCR扩增产物进行测序。
GP2027-2018-F:5’-GTGGAGTTCTCGTACCCGTC-3’;
GP2027-2018-R:5’-TTGACGATGAAGGAGTGGGC-3’。
通过以上鉴定,从步骤二得到的再生植株中筛选得到两个纯合突变型植株(即两条染色体发生的突变一致),分别命名为Osabcb1-4植株和Osabcb1-7植株。
经测序鉴定,与水稻日本晴(用Nip表示)的基因组DNA相比,Osabcb1-4植株的差异仅在于在编码OsABCB1蛋白的基因中发生了一个核苷酸的插入,该突变位点及其周边核苷酸的测序结果见图2。
经测序鉴定,与水稻日本晴(用Nip表示)的基因组DNA相比,Osabcb1-7植株的差异仅在于在编码OsABCB1蛋白的基因中发生了两个核苷酸的缺失,该突变位点及其周边核苷酸的测序结果见图2。
Osabcb1-4植株自交并收获种子,将种子培育为植株,即为T1代植株。T1代植株自交并收获种子,将种子培育为植株,即为T2代植株。Osabcb1-4植株及其自交后代,称为Osabcb1-4株系。
Osabcb1-7植株自交并收获种子,将种子培育为植株,即为T1代植株。T1代植株自交并收获种子,将种子培育为植株,即为T2代植株。Osabcb1-7植株及其自交后代,称为Osabcb1-7株系。
四、性状比较
供试植株为:水稻日本晴(用Nip表示)、Osabcb1-4株系的T2代植株、Osabcb1-7株系的T2代植株。
在平行条件下培养供试植株。
成熟期植株的照片见图3。成熟期植株的株高见图4(每个株系取10株植株的平均值)。与水稻日本晴相比,Osabcb1-4株系植株和Osabcb1-7株系植株均株高显著降低。Osabcb1-4株系植株和Osabcb1-7株系植株的株高无显著差异。与水稻日本晴相比,Osabcb1-4株系植株和Osabcb1-7株系植株均叶夹角显著变小。Osabcb1-4株系植株和Osabcb1-7株系植株的叶夹角无显著差异。
成熟稻穗的照片见图5。成熟稻穗的穗长见图6(每个株系取10株植株的平均值)。水稻日本晴、Osabcb1-4株系植株和Osabcb1-7株系植株的穗长均无显著差异。
成熟籽粒的照片见图7(上图为未脱壳籽粒,下图为脱壳籽粒)。千粒重的结果见图8。粒长和粒宽的结果见图9(每个株系取10株植株的平均值)。水稻日本晴、Osabcb1-4株系植株和Osabcb1-7株系植株的粒长均无显著差异。水稻日本晴、Osabcb1-4株系植株和Osabcb1-7株系植株的千粒重均无显著差异。水稻日本晴、Osabcb1-4株系植株和Osabcb1-7株系植株的粒宽均无显著差异。
以上各个性状的数据结果见表2。
表2
水稻日本晴 Osabcb1-4株系植株 Osabcb1-7株系植株
株高 72.55cm 53.45cm 52.98cm
穗长 17.53cm 16.71cm 16.80cm
粒长 0.68cm 0.66cm 0.67cm
粒宽 0.32cm 0.32cm 0.33cm
千粒重 20.92g 20.89g 20.91g
SEQUENCE LISTING
<110> 中国农业科学院生物技术研究所
<120> OsABCB1蛋白及其编码基因和应用
<130> GNCYX191204
<160> 6
<170> PatentIn version 3.5
<210> 1
<211> 1344
<212> PRT
<213> Oryza sativa
<400> 1
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Val Cys Gly Ser Phe Ser Ala Ile Phe Ala Tyr Val Leu Ser Ala Val
770 775 780
Leu Ser Val Tyr Tyr Ala Pro Asp Ala Ala Tyr Met Asp Arg Gln Ile
785 790 795 800
Ala Lys Tyr Cys Tyr Leu Leu Ile Gly Met Ser Ser Ala Ala Leu Leu
805 810 815
Phe Asn Thr Val Gln His Leu Phe Trp Asp Thr Val Gly Glu Asn Leu
820 825 830
Thr Lys Arg Val Arg Glu Arg Met Leu Ala Ala Val Leu Arg Asn Glu
835 840 845
Ile Ala Trp Phe Asp Met Glu Asp Asn Ser Ser Ala Arg Ile Ala Ala
850 855 860
Arg Leu Ala Leu Asp Ala Gln Asn Val Arg Ser Ala Ile Gly Asp Arg
865 870 875 880
Ile Ser Ile Ile Val Gln Asn Ser Ala Leu Met Leu Val Ala Cys Thr
885 890 895
Ala Gly Phe Val Leu Gln Trp Arg Leu Ala Leu Val Leu Leu Ala Val
900 905 910
Phe Pro Leu Val Val Ala Ala Thr Val Leu Gln Lys Met Phe Leu Lys
915 920 925
Gly Phe Ser Gly Asp Leu Glu Arg Ala His Ala Arg Ala Thr Gln Ile
930 935 940
Ala Gly Glu Ala Val Ala Asn Val Arg Thr Val Ala Ala Phe Gly Ser
945 950 955 960
Glu Ala Lys Ile Val Gly Leu Phe Glu Ala Asn Leu Ala Gly Pro Leu
965 970 975
Arg Arg Cys Phe Trp Lys Gly Gln Ile Ala Gly Ser Gly Tyr Gly Val
980 985 990
Ala Gln Phe Leu Leu Tyr Ala Ser Tyr Ala Leu Gly Leu Trp Tyr Ala
995 1000 1005
Ala Trp Leu Val Lys His Gly Val Ser Asp Phe Ser Lys Thr Ile
1010 1015 1020
Arg Val Phe Met Val Leu Met Val Ser Ala Asn Gly Ala Ala Glu
1025 1030 1035
Thr Leu Thr Leu Ala Pro Asp Phe Val Lys Gly Gly Arg Ala Met
1040 1045 1050
Gln Ala Val Phe Glu Ala Met Asp Arg Arg Thr Glu Ile Glu Pro
1055 1060 1065
Asp Asp Val Asp Ala Ala Ala Val Pro Glu Arg Pro Arg Gly Glu
1070 1075 1080
Val Glu Leu Lys His Val Asp Phe Ala Tyr Pro Ser Arg Pro Glu
1085 1090 1095
Val Gln Val Phe Arg Asp Leu Ser Leu Arg Ala Arg Ala Gly Arg
1100 1105 1110
Thr Leu Ala Leu Val Gly Ala Ser Gly Cys Gly Lys Ser Ser Val
1115 1120 1125
Leu Ala Leu Val Gln Arg Phe Tyr Glu Pro Asn Ser Gly Arg Val
1130 1135 1140
Leu Leu Asp Gly Arg Asp Leu Arg Lys Phe Asn Leu Arg Ser Leu
1145 1150 1155
Arg Arg Ala Met Ala Leu Val Pro Gln Glu Pro Phe Leu Phe Ala
1160 1165 1170
Ala Thr Ile His Asp Asn Ile Ala Tyr Gly Arg Glu Gly Ala Thr
1175 1180 1185
Glu Ala Glu Val Val Glu Ala Ala Thr Ala Ala Asn Ala His Lys
1190 1195 1200
Phe Ile Ser Ala Leu Pro Glu Gly Tyr Gly Thr Leu Val Gly Glu
1205 1210 1215
Arg Gly Val Gln Leu Ser Gly Gly Gln Arg Gln Arg Ile Ala Ile
1220 1225 1230
Ala Arg Ala Leu Val Lys Gln Ala Pro Ile Leu Leu Leu Asp Glu
1235 1240 1245
Ala Thr Ser Ala Leu Asp Ala Glu Ser Glu Arg Ser Val Gln Glu
1250 1255 1260
Ala Leu Ala Ser Ser Ser Gly Ser Gly Arg Thr Thr Ile Val Val
1265 1270 1275
Ala His Arg Leu Ala Thr Val Arg Asn Ala His Thr Ile Ala Val
1280 1285 1290
Ile Asp Asp Gly Lys Val Ala Glu Gln Gly Ser His Ser His Leu
1295 1300 1305
Leu Asn His His Pro Asp Gly Cys Tyr Ala Arg Met Leu Gln Leu
1310 1315 1320
Gln Arg Leu Ser His Ser His Val Ala Pro Gly Pro Ser Ser Ser
1325 1330 1335
Thr Thr Thr His Gly Thr
1340
<210> 2
<211> 4035
<212> DNA
<213> Oryza sativa
<400> 2
atggaggagg agataaaggg ccgcgtcgtc gtcctcggcg ccgatgccgc cgccgacccg 60
gagctcgagg ccttccatct cccctccgcc gaccagcccc cgcactcgca cctcctccac 120
caccaccatt ctccacaatc ccatcctcaa cctgatgcac cagcagcagc agcgccaccg 180
ccacctgctc cccttactcc tcctcctcct aagtcgccgc cgccgcctcc ccacatccaa 240
accaccgacc ttccaccgcc caagcccctc ccgccggccc cgctccgtca actcttcagc 300
ttcgccgacg gcctcgacta cgtcctcatg accctgggca cgctgggcgc cctcgtccac 360
ggctgctccc tcccggtctt cctccgcttc ttcgccgacc tcgtcgactc cttcggctcc 420
cacgccgccc acccggacac catgctccgc ctcgtcgtca agtacgcctt ctacttcctc 480
gtcgtcggcg ccgccatctg ggcctcctcc tgggccgaga tctcctgctg gatgtggacc 540
ggcgagcggc agtccacccg gatgcgcatc cgctacctcc acgccgccct ccaccaggac 600
gtctccttct tcgacaccga cgtccgcacc tccgacgtca tccacgccat caacgccgac 660
gctgtcgtcg tccaggacgc catcagcgag aagctcggca acctcatcca ctacctcgcc 720
accttcgtct ccggcttcgt cgtcggattc accgccgcct ggcagctcgc cctcgtcacc 780
ctcgccgtcg tcccgctcat cgccgtcatc ggcggcctca gcgccgccgc gctcgccaag 840
ctctcctcca ggagccagga cgcgctgtcc gacgccagcg gcatcgcgga gcaggcgctg 900
gcgcagatac gcatcgtgca gtcgttcgtg ggcgaggagc gggtgatgcg ggcctactcg 960
gcggcgctgg ccgtggcgca gcggatcggc taccgctccg gcttcgccaa gggcatcgga 1020
ctgggcggca cctacttcac cgtcttctgc tgctacgcgc tgctcctctg gtacgggggt 1080
cacctcgtcc gacgcgccca caccaacgga ggcctcgcca tcgccaccat gttctcagta 1140
atgataggcg gcctggctct tgggcagtcg gctccgagca tggcggcgtt cgccaaggcg 1200
cgggtggcgg cggcgaagat attccggatg atggagcaca agccgagcat ggagcgggag 1260
ggcggggtgg agctggaggc ggtgacgggg cgggtggagc tgcgggatgt ggagttctcg 1320
tacccgtcgc ggccagacgt ggggatcctg cgggggttgt cgctgagcgt gccggcgggg 1380
aagacgatcg cgctggtggg gagctccggg tcagggaaga gcacggtggt gtcgctgatc 1440
gagaggttct acgagcccaa cgccgggacc atcctgctgg acggccacga cctgagggac 1500
ctcaacctgc ggtggctgcg ccgccagatt ggtctggtga gccaggagcc ggcgctgttc 1560
gccaccacca tccgggagaa cctgctgctg gggagggacg gcgccacgca ggaggagttg 1620
gaggaggccg ccagggtcgc caacgcccac tccttcatcg tcaagctccc cgacgcctac 1680
aacacgcagg ttggggagcg cgggctgcag ttgtcgggcg ggcagaagca gcggatcgcc 1740
atcgcgcgcg ccatgctccg gaacccggcc atcctgctgc tggacgaggc caccagcgcg 1800
ctggactcgg agtcggagaa gctggtgcag gaggcgctgg accgcttcat gatcgggcgc 1860
accacgctgg tgatcgcgca ccgcctctcc accatccgca aggccgacct cgtggccgtc 1920
ctccagggcg gggccatctc cgaggtgggc acgcacgacg agctcatggc ccgcggggat 1980
ggcacctacg ccaggctcat ccgaatgcag gagcaggcgc acgaggccgc gctcgtcgcc 2040
gctcgccgga gcagcgcgcg accatccagc gcccgcaact ccgtcagctc gcccatcatc 2100
acgcgtaact cctcctacgg ccgttccccc tactcccgcc gcctctccga cgccgacttc 2160
atcaccggcc tcggcctcgg agtcgacagc aagcagcagc agcagcagca ctacttccgc 2220
gtgcaggcca gctccttctg gcggctggcc aagatgaact cgcccgagtg ggggtacgcg 2280
ctcgtcgcgt cgctgggctc catggtgtgc ggctccttca gcgccatctt cgcctacgtc 2340
ctcagcgccg tgctcagcgt gtactacgcc cccgacgccg cctacatgga ccgccagatc 2400
gccaagtact gctacctcct catcggcatg tcctccgccg ccctgctctt caacactgtc 2460
cagcacctct tctgggacac cgtcggcgag aacctcacca agcgcgtccg cgagaggatg 2520
ctcgccgccg tgctccgcaa cgagatcgcc tggttcgaca tggaggacaa ctccagcgcc 2580
cgcatcgccg ccaggctggc gctggacgcc cagaacgtgc gctccgccat cggcgaccgc 2640
atctccatca tcgtccagaa ctcggcgctc atgctggtgg cctgcaccgc cggcttcgtc 2700
ctgcagtggc gcctcgcgct ggtgctgctg gccgtcttcc cgctggtggt ggccgccacc 2760
gttctgcaga agatgttcct caaggggttc tcgggcgacc tggagcgggc gcacgcgagg 2820
gcgacgcaga tcgcgggaga ggcggtggcg aacgtgcgca ccgtggcggc gttcgggtcg 2880
gaggcgaaga tcgtggggct gttcgaggcg aacctggcgg ggccgctccg gcggtgcttc 2940
tggaaggggc agatcgccgg gagcgggtac ggggtggcgc agttcctgct gtacgcgtcg 3000
tacgcgctgg ggctgtggta cgcggcgtgg ctggtgaagc acggcgtgtc ggacttctcc 3060
aagaccatcc gggtgttcat ggtgctgatg gtgtcggcca acggcgccgc ggagacgctg 3120
acgctggcgc cggacttcgt gaagggcggg cgggcgatgc aggcggtgtt cgaggcgatg 3180
gaccggagga cagagatcga gccggacgac gtggacgcgg cggcggtgcc ggagcggccc 3240
cggggggagg ttgagctgaa gcacgtggac ttcgcgtacc cgtcgcggcc ggaggtgcag 3300
gtgttccggg acctgagcct gagggcgcgg gcggggcgga cgctggcgct ggtgggggcg 3360
agcgggtgcg gcaagagctc ggtgctggcg ctggtgcagc gcttctacga gcccaactcc 3420
gggcgggtgc tcctggacgg ccgggacctg cgcaagttca acctgcggtc cctgcgtcgc 3480
gccatggcgc tggtgccgca ggagcccttc cttttcgcgg ccaccatcca cgacaacatc 3540
gcgtacgggc gcgagggggc gacggaggcg gaggtggtgg aggcggcgac ggcggccaac 3600
gcgcacaagt tcatatcggc gctgccggag gggtacggga cgttggtcgg ggagcgcggg 3660
gtgcagctgt cgggcgggca gcgccagcgg atcgccatcg cgcgggcgct ggtgaagcag 3720
gcgcccatcc tgctgctgga cgaggccacc agcgcgctgg acgccgagtc ggagcggagc 3780
gtgcaggagg cgctcgcctc ctcctctggc tccggccgca ccaccatcgt ggtggcccac 3840
cgtctggcca ccgtccgcaa cgcccacacc atcgccgtca tcgacgacgg caaggtggcg 3900
gagcagggct cgcactccca cctgctcaac caccaccccg atggctgcta cgcgcgcatg 3960
ctgcagctcc agcgcctctc ccactcccac gttgcgccgg gcccctcctc ctcaacaaca 4020
acacacggca cctag 4035
<210> 3
<211> 8260
<212> DNA
<213> Oryza sativa
<400> 3
ttgttgagga gtcgttgcca ttggtgagtc ttctccctct tccttgcttg gagtttgacg 60
acgattcctt gagaaaggtg acaggggaat cttaatcata tgagaactgg caaatccctt 120
agtaaggagc agacccaaac cgccatgacc catctggtgt tgaaagccca tgggtcgaga 180
ctcttttttc accatgacgg gttgagatga tgaaaaatta gcataggccg acaccagctt 240
tgtctctagg ctggatttgc ccctgtcccc agtagttact tacttttttt taatctaatc 300
ttttctttat tttttttgag ttgtcaaatt aaatgtgcta ctatagtaag tatatagtac 360
agtaaataaa tgttggttac ttatgattgt gcgtacaagc catgtaatga cttaccgcaa 420
tgccgtccgt gcaatgcgtg agcggaatgg attggggtgc gataagatgg ttgacgacat 480
tgttgtaatg gtacaacatt gttagagatc acgtggttcc ataagataag taaagcgatt 540
ggccacataa cattttctcc gttggcgtga atcagcgtaa gttgcaagca tttggctggc 600
cgtgcatcac ctgtggcata cagttacaat agtaattaat taaattagtt actaccatcg 660
taccataaag taggagtata aaattaccac accatgcacc atggtccatg ggtgtttgtg 720
atgaaaatat atctttttag aaagaaatac tactagtata agctacttgg gggtttgttt 780
taaaattgaa gaaagaaagt ttgcaggaat ggtttggtgc gagagagaga tataggtgta 840
gatcgatatg tacgggtgga cttggtgact gtggcagtta gcataagtat tggaagaatt 900
ttttcttaaa aaaaaaagat ttgagactgc tggccggtcg ggcggcacct agctagcaac 960
tctgtctgca ctctgtgtag gtgttgaagg tttgattgga ggaggagtag tactagtagt 1020
attggccgtg cttagattta aacttttttt taaaactttt aacttttccg tcatattaaa 1080
tgtttggaca catgcataga gcattaaata tggattaaaa aaatcaattg cacggtttgt 1140
atgtaaatcg taagatgaat ttttgagcct aattacgcca tgatttgaca atttgatgct 1200
acagtaaata ttgactaatg acggattaat taggcttaat cgattcgtct cacagtttat 1260
aggtgaaatc tataattgat tttattatta atctacgttt aataatttaa atgtgtgtcc 1320
gtatacttaa aaagaatttt ggcacacgaa ctatgcacag ccatgcttag gagtaccgat 1380
cgagtacgta atagggagta acctctccgt ttcgaaatgt ctgacgccgt tgacttttta 1440
gtacatgttt gaccgttcgt cttattcaaa aaatttaagt aattattaat tctttttata 1500
tcatttgatt cattgttaaa tatattttta tgtaggcata taattttaca tatttcacaa 1560
aagtttttga ataagacaaa cggtcaaata tttactaaaa agtcaacggt gtcaaatatt 1620
tcaaaacaga gggagtatat actagcagct agtccagcat gcgcatacat gcatactagt 1680
actccagtac gagttgtaca aatgcgtaca gtgaaaatgg agtgcgacaa catgtgcata 1740
ctactacttc ttacgtagca tagtgatttg tgacagtggt cgtcggtcgt cgttggaaat 1800
aaaagcttga ttatattgtc aagtgaagca tgtggtaggt tttggtttgg atgattggta 1860
gttagctcgt gtgaactctg aagcaacaat atgtgaaagg accaactgca acatcctgcc 1920
tgcctgcgga tattatatta ccgtggaaag agagagagag attaccggag ctccgagctt 1980
ggggctccgg gcgggtcggc cggtgaggtt gaaggaggcc acacacaccc ggtaagagta 2040
ttaccgtgga taactactac tctacatgaa gggaaaaagg gctggctggt gtagaactag 2100
aagccgactt ggtgtggcct agtggaccag gcttgttgtg caaggccacg tgccttttct 2160
tttctattag tccagtagtc tctctattta taaacatgtc gtcccctctt accaccagcc 2220
ataagccatt aggcaaggca atcctcctcc tcctccttgc tcacagtcac agatgggaag 2280
agaggaattg agcaactcgt caactcaaga attggagggt tgagctagaa ttattaggaa 2340
gtagagctag aattagagaa gagcagagca gcaagagatt attagtacag gaacaggaga 2400
gcagagcaga gtgcgatcga gcaagggcag ggcggcggta ggctaggcat ggaggaggag 2460
ataaagggcc gcgtcgtcgt cctcggcgcc gatgccgccg ccgacccgga gctcgaggcc 2520
ttccatctcc cctccgccga ccagcccccg cactcgcacc tcctccacca ccaccattct 2580
ccacaatccc atcctcaacc tgatgcacca gcagcagcag cgccaccgcc acctgctccc 2640
cttactcctc ctcctcctaa gtcgccgccg ccgcctcccc acatccaaac caccgacctt 2700
ccaccgccca agcccctccc gccggccccg ctccgtcaac tcttcagctt cgccgacggc 2760
ctcgactacg tcctcatgac cctgggcacg ctgggcgccc tcgtccacgg ctgctccctc 2820
ccggtcttcc tccgcttctt cgccgacctc gtcgactcct tcggctccca cgccgcccac 2880
ccggacacca tgctccgcct cgtcgtcaag tacgccttct acttcctcgt cgtcggcgcc 2940
gccatctggg cctcctcctg ggccgagatc tcctgctgga tgtggaccgg cgagcggcag 3000
tccacccgga tgcgcatccg ctacctccac gccgccctcc accaggacgt ctccttcttc 3060
gacaccgacg tccgcacctc cgacgtcatc cacgccatca acgccgacgc tgtcgtcgtc 3120
caggacgcca tcagcgagaa gctcggcaac ctcatccact acctcgccac cttcgtctcc 3180
ggcttcgtcg tcggattcac cgccgcctgg cagctcgccc tcgtcaccct cgccgtcgtc 3240
ccgctcatcg ccgtcatcgg cggcctcagc gccgccgcgc tcgccaagct ctcctccagg 3300
agccaggacg cgctgtccga cgccagcggc atcgcggagc aggcgctggc gcagatacgc 3360
atcgtgcagt cgttcgtggg cgaggagcgg gtgatgcggg cctactcggc ggcgctggcc 3420
gtggcgcagc ggatcggcta ccgctccggc ttcgccaagg gcatcggact gggcggcacc 3480
tacttcaccg tcttctgctg ctacgcgctg ctcctctggt acgggggtca cctcgtccga 3540
cgcgcccaca ccaacggagg cctcgccatc gccaccatgt tctcagtaat gataggcggc 3600
ctgtaagtaa agctttgtag tactccgtag catatacgta ctccgattgg ccggattgag 3660
agtaaaaaca aaacaaaaat ccaatccgat ccgcagggct cttgggcagt cggctccgag 3720
catggcggcg ttcgccaagg cgcgggtggc ggcggcgaag atattccgga tgatggagca 3780
caagccgagc atggagcggg agggcggggt ggagctggag gcggtgacgg ggcgggtgga 3840
gctgcgggat gtggagttct cgtacccgtc gcggccagac gtggggatcc tgcgggggtt 3900
gtcgctgagc gtgccggcgg ggaagacgat cgcgctggtg gggagctccg ggtcagggaa 3960
gagcacggtg gtgtcgctga tcgagaggtt ctacgagccc aacgccggga ccatcctgct 4020
ggacggccac gacctgaggg acctcaacct gcggtggctg cgccgccaga ttggtctggt 4080
gagccaggag ccggcgctgt tcgccaccac catccgggag aacctgctgc tggggaggga 4140
cggcgccacg caggaggagt tggaggaggc cgccagggtc gccaacgccc actccttcat 4200
cgtcaagctc cccgacgcct acaacacgca ggcaagtatt ttgctgcttc cgtcaatttc 4260
cttcgtttca cttgttgttg ttgttgctgt ttgtccttcc tgtcgctgcc tctcctaacc 4320
ctgtgagacg atgacgatcc accatgcaac aatgcgttgt gtttgtttgt taggcaggca 4380
gcatgagcac gaggaggcat gctatgctat cacattcaca ccacgtctct ctgctgcttc 4440
tcatatcatc tcatctccct cctttttagt tatagataga tagtacatta gagattactg 4500
tactcggagg gagagagata ctcctagata gagacaggga aagacatgtt atgttattat 4560
tactagggcc aggcggcaat caatgggttg ctttcgaacg ctgttccgag ttggtgcaag 4620
taacatggac ggattattgg atcatgccaa gattcctcct cctcttcctc cccctttcaa 4680
aagcacaaac catggcacct gcacactcat caccttcttt cgacttctcc atctacaaca 4740
gctacagtac ctactaccac tacggactag tactagatca gaagatgtga cgcccttttg 4800
cagcacccac tcatcaactg cacttgtcca ctccagtgcc cactgcagct cattgccaaa 4860
tttcaaggaa tcctaagctc gggatgattc cattccagct tgacattttg tcaggaatgt 4920
cagcccgctt cttcctccca ataatcaatc ctttgttttc ttttcttttc ttttctttcc 4980
ctcttgtttg atggggctcg ccgctcgctc atggagtcat gagctcttgc caggcggttt 5040
gcttaattaa accttaacac tccggccctt cattaattat tcttgccagc tgagtagctg 5100
acacgcctgc ctgctgcatt agcatttagt aacaggggga gtagtagtag gttagttaca 5160
cagtagtgga gtactccaga atattgttgt tctcgttctg ctgcgcacat gacacggtag 5220
gcagtagtat ggcacccatg aatatgatgc tttgcttaac accattacta attaattacc 5280
ttgacaagta caataaattg aatgagttga acaccaagca agaaattaac gagtagtaag 5340
cgaatgttta ggctgacgca aaaagcatgg aatggcaatg caggttgggg agcgcgggct 5400
gcagttgtcg ggcgggcaga agcagcggat cgccatcgcg cgcgccatgc tccggaaccc 5460
ggccatcctg ctgctggacg aggccaccag cgcgctggac tcggagtcgg agaagctggt 5520
gcaggaggcg ctggaccgct tcatgatcgg gcgcaccacg ctggtgatcg cgcaccgcct 5580
ctccaccatc cgcaaggccg acctcgtggc cgtcctccag ggcggggcca tctccgaggt 5640
gggcacgcac gacgagctca tggcccgcgg ggatggcacc tacgccaggc tcatccgaat 5700
gcaggagcag gcgcacgagg ccgcgctcgt cgccgctcgc cggagcagcg cgcgaccatc 5760
cagcgcccgc aactccgtca gctcgcccat catcacgcgt aactcctcct acggccgttc 5820
cccctactcc cgccgcctct ccgacgccga cttcatcacc ggcctcggcc tcggagtcga 5880
cagcaagcag cagcagcagc agcactactt ccgcgtgcag gccagctcct tctggcggct 5940
ggccaagatg aactcgcccg agtgggggta cgcgctcgtc gcgtcgctgg gctccatggt 6000
gtgcggctcc ttcagcgcca tcttcgccta cgtcctcagc gccgtgctca gcgtgtacta 6060
cgcccccgac gccgcctaca tggaccgcca gatcgccaag tactgctacc tcctcatcgg 6120
catgtcctcc gccgccctgc tcttcaacac tgtccagcac ctcttctggg acaccgtcgg 6180
cgagaacctc accaagcgcg tccgcgagag gatgctcgcc gccgtgctcc gcaacgagat 6240
cgcctggttc gacatggagg acaactccag cgcccgcatc gccgccaggc tggcgctgga 6300
cgcccagaac gtgcgctccg ccatcggcga ccgcatctcc atcatcgtcc agaactcggc 6360
gctcatgctg gtggcctgca ccgccggctt cgtcctgcag tggcgcctcg cgctggtgct 6420
gctggccgtc ttcccgctgg tggtggccgc caccgttctg cagaagatgt tcctcaaggg 6480
gttctcgggc gacctggagc gggcgcacgc gagggcgacg cagatcgcgg gagaggcggt 6540
ggcgaacgtg cgcaccgtgg cggcgttcgg gtcggaggcg aagatcgtgg ggctgttcga 6600
ggcgaacctg gcggggccgc tccggcggtg cttctggaag gggcagatcg ccgggagcgg 6660
gtacggggtg gcgcagttcc tgctgtacgc gtcgtacgcg ctggggctgt ggtacgcggc 6720
gtggctggtg aagcacggcg tgtcggactt ctccaagacc atccgggtgt tcatggtgct 6780
gatggtgtcg gccaacggcg ccgcggagac gctgacgctg gcgccggact tcgtgaaggg 6840
cgggcgggcg atgcaggcgg tgttcgaggc gatggaccgg aggacagaga tcgagccgga 6900
cgacgtggac gcggcggcgg tgccggagcg gccccggggg gaggttgagc tgaagcacgt 6960
ggacttcgcg tacccgtcgc ggccggaggt gcaggtgttc cgggacctga gcctgagggc 7020
gcgggcgggg cggacgctgg cgctggtggg ggcgagcggg tgcggcaaga gctcggtgct 7080
ggcgctggtg cagcgcttct acgagcccaa ctccgggcgg gtgctcctgg acggccggga 7140
cctgcgcaag ttcaacctgc ggtccctgcg tcgcgccatg gcgctggtgc cgcaggagcc 7200
cttccttttc gcggccacca tccacgacaa catcgcgtac gggcgcgagg gggcgacgga 7260
ggcggaggtg gtggaggcgg cgacggcggc caacgcgcac aagttcatat cggcgctgcc 7320
ggaggggtac gggacgttgg tcggggagcg cggggtgcag ctgtcgggcg ggcagcgcca 7380
gcggatcgcc atcgcgcggg cgctggtgaa gcaggcgccc atcctgctgc tggacgaggc 7440
caccagcgcg ctggacgccg agtcggagcg gagcgtgcag gaggcgctcg cctcctcctc 7500
tggctccggc cgcaccacca tcgtggtggc ccaccgtctg gccaccgtcc gcaacgccca 7560
caccatcgcc gtcatcgacg acggcaaggt ggcggagcag ggctcgcact cccacctgct 7620
caaccaccac cccgatggct gctacgcgcg catgctgcag ctccagcgcc tctcccactc 7680
ccacgttgcg ccgggcccct cctcctcaac aacaacacac ggcacctagt attcgttttg 7740
atacaacaac atgttcttct tttgctgtgt gttcttgctg ctgctgctgc tagatgttct 7800
aagaatgaat taatgaaatg aattactata ctgctagcta gctagctcat gatactacta 7860
gtactgtgtc catcgatgga tcattttagc tcgattcttc gattagtttg tccggaaatc 7920
aaggaccagc acaaaagcag catgctcagt tcgattagtt gaggatgatt gatacaccac 7980
cactgctgct gctgctgctg ctggagtagt ataatattac tccattacca agtcaattcg 8040
atcagcaggt atgacccggg atttagggct tccttgtcac tactactagt acaacattca 8100
gagtagtcgt aggagtagga gtatcattcg ttcacactcc aaagtccaaa gtaacagtgt 8160
tttgcaggca agggataggg agggagggag gcagaattgc agaaattctg gaccctccaa 8220
agtttgactt ggaaggacta tcacatgcac atgcaggcat 8260
<210> 4
<211> 15898
<212> DNA
<213> Artificial sequence
<400> 4
taaacgctct tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta 60
aactgaaggc gggaaacgac aatctgatcc aagctcaagc tgctctagca ttcgccattc 120
aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 180
gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca 240
cgacgttgta aaacgacggc cagtgccaag cttggatcat gaaccaacgg cctggctgta 300
tttggtggtt gtgtagggag atggggagaa gaaaagcccg attctcttcg ctgtgatggg 360
ctggatgcat gcgggggagc gggaggccca agtacgtgca cggtgagcgg cccacagggc 420
gagtgtgagc gcgagaggcg ggaggaacag tttagtacca cattgcccag ctaactcgaa 480
cgcgaccaac ttataaaccc gcgcgctgtc gcttgtgtgc tggacggcca cgacctgagt 540
tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg 600
caccgagtcg gtgctttttt gttttagagc tagaaatagc aagttaaaat aaggctagtc 660
cgtagcgcgt gcgccaattc tgcagacaaa tggccccggg cctgcaggtg cagcgtgacc 720
cggtcgtgcc cctctctaga gataatgagc attgcatgtc taagttataa aaaattacca 780
catatttttt ttgtcacact tgtttgaagt gcagtttatc tatctttata catatattta 840
aactttactc tacgaataat ataatctata gtactacaat aatatcagtg ttttagagaa 900
tcatataaat gaacagttag acatggtcta aaggacaatt gagtattttg acaacaggac 960
tctacagttt tatcttttta gtgtgcatgt gttctccttt ttttttgcaa atagcttcac 1020
ctatataata cttcatccat tttattagta catccattta gggtttaggg ttaatggttt 1080
ttatagacta atttttttag tacatctatt ttattctatt ttagcctcta aattaagaaa 1140
actaaaactc tattttagtt tttttattta ataatttaga tataaaatag aataaaataa 1200
agtgactaaa aattaaacaa atacccttta agaaattaaa aaaactaagg aaacattttt 1260
cttgtttcga gtagataatg ccagcctgtt aaacgccgtc gacgagtcta acggacacca 1320
accagcgaac cagcagcgtc gcgtcgggcc aagcgaagca gacggcacgg catctctgtc 1380
gctgcctctg gacccctctc gagagttccg ctccaccgtt ggacttgctc cgctgtcggc 1440
atccagaaat tgcgtggcgg agcggcagac gtgagccggc acggcaggcg gcctcctcct 1500
cctctcacgg cacggcagct acgggggatt cctttcccac cgctccttcg ctttcccttc 1560
ctcgcccgcc gtaataaata gacaccccct ccacaccctc tttccccaac ctcgtgttgt 1620
tcggagcgca cacacacaca accagatctc ccccaaatcc acccgtcggc acctccgctt 1680
caaggtacgc cgctcgtcct cccccccccc ccctctctac cttctctaga tcggcgttcc 1740
ggtccatggt tagggcccgg tagttctact tctgttcatg tttgtgttag atccgtgttt 1800
gtgttagatc cgtgctgcta gcgttcgtac acggatgcga cctgtacgtc agacacgttc 1860
tgattgctaa cttgccagtg tttctctttg gggaatcctg ggatggctct agccgttccg 1920
cagacgggat cgatttcatg attttttttg tttcgttgca tagggtttgg tttgcccttt 1980
tcctttattt caatatatgc cgtgcacttg tttgtcgggt catcttttca tgcttttttt 2040
tgtcttggtt gtgatgatgt ggtctggttg ggcggtcgtt ctagatcgga gtagaattct 2100
gtttcaaact acctggtgga tttattaatt ttggatctgt atgtgtgtgc catacatatt 2160
catagttacg aattgaagat gatggatgga aatatcgatc taggataggt atacatgttg 2220
atgcgggttt tactgatgca tatacagaga tgctttttgt tcgcttggtt gtgatgatgt 2280
ggtgtggttg ggcggtcgtt cattcgttct agatcggagt agaatactgt ttcaaactac 2340
ctggtgtatt tattaatttt ggaactgtat gtgtgtgtca tacatcttca tagttacgag 2400
tttaagatgg atggaaatat cgatctagga taggtataca tgttgatgtg ggttttactg 2460
atgcatatac atgatggcat atgcagcatc tattcatatg ctctaacctt gagtacctat 2520
ctattataat aaacaagtat gttttataat tattttgatc ttgatatact tggatgatgg 2580
catatgcagc agctatatgt ggattttttt agccctgcct tcatacgcta tttatttgct 2640
tggtactgtt tcttttgtcg atgctcaccc tgttgtttgg tgttacttct gcagccatgg 2700
actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg 2760
ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca 2820
agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg 2880
acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca 2940
tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc 3000
ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc 3060
aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag 3120
agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg 3180
tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg 3240
tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca 3300
agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca 3360
agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg 3420
ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg 3480
aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg 3540
ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca 3600
aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg 3660
gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga 3720
gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca 3780
agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc 3840
tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca 3900
ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga 3960
tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc 4020
ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc 4080
tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga 4140
tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg 4200
cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg 4260
acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc 4320
ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg 4380
agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg 4440
agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc 4500
agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg 4560
tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg 4620
acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc 4680
tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt 4740
tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga 4800
gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc 4860
tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga 4920
cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc 4980
acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg 5040
tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa 5100
tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc 5160
ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa 5220
acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt 5280
acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc 5340
ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga 5400
accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact 5460
ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg 5520
ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg 5580
aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt 5640
acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg 5700
tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc 5760
acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta 5820
agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg 5880
ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca 5940
tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga 6000
tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg 6060
tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag 6120
gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa 6180
agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg 6240
tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc 6300
tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg 6360
aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc 6420
tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg 6480
gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg 6540
agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca 6600
agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg 6660
ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca 6720
gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg 6780
ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc 6840
tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt 6900
ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga 6960
aaaagtaagg atcctgattg atcgatagag ctcgaatttc cccgatcgtt caaacatttg 7020
gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt 7080
tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag 7140
atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat 7200
atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcggga 7260
attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 7320
acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 7380
tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 7440
tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggc tagagcagct 7500
tgccaacatg gtggagcacg acactctcgt ctactccaag aatatcaaag atacagtctc 7560
agaagaccaa agggctattg agacttttca acaaagggta atatcgggaa acctcctcgg 7620
attccattgc ccagctatct gtcacttcat caaaaggaca gtagaaaagg aaggtggcac 7680
ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt caagatgcct ctgccgacag 7740
tggtcccaaa gatggacccc cacccacgag gagcatcgtg gaaaaagaag acgttccaac 7800
cacgtcttca aagcaagtgg attgatgtga taacatggtg gagcacgaca ctctcgtcta 7860
ctccaagaat atcaaagata cagtctcaga agaccaaagg gctattgaga cttttcaaca 7920
aagggtaata tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcaa 7980
aaggacagta gaaaaggaag gtggcaccta caaatgccat cattgcgata aaggaaaggc 8040
tatcgttcaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag 8100
catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt gatgtgatat 8160
ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc ttcctctata 8220
taaggaagtt catttcattt ggagaggaca cgctgaaatc accagtctct ctctacaaat 8280
ctatctctct cgagctttcg cagatcccgg ggggcaatga gatatgaaaa agcctgaact 8340
caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat 8400
gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata 8460
tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca 8520
ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt ttagcgagag 8580
cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac 8640
cgaactgccc gctgttctac aaccggtcgc ggaggctatg gatgcgatcg ctgcggccga 8700
tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac 8760
atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat 8820
ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga 8880
ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac 8940
ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca 9000
atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac 9060
gcgctacttc gagcggaggc atccggagct tgcaggatcg ccacgactcc gggcgtatat 9120
gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc 9180
agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg 9240
tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc 9300
cgatagtgga aaccgacgcc ccagcactcg tccgagggca aagaaataga gtagatgccg 9360
accggatctg tcgatcgaca agctcgagtt tctccataat aatgtgtgag tagttcccag 9420
ataagggaat tagggttcct atagggtttc gctcatgtgt tgagcatata agaaaccctt 9480
agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa 9540
aatccagtac taaaatccag atcccccgaa ttaattcggc gttaattcag tacattaaaa 9600
acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt gtttacacca caatatatcc 9660
tgccaccagc cagccaacag ctccccgacc ggcagctcgg cacaaaatca ccactcgata 9720
caggcagccc atcagtccgg gacggcgtca gcgggagagc cgttgtaagg cggcagactt 9780
tgctcatgtt accgatgcta ttcggaagaa cggcaactaa gctgccgggt ttgaaacacg 9840
gatgatctcg cggagggtag catgttgatt gtaacgatga cagagcgttg ctgcctgtga 9900
tcaccgcggt ttcaaaatcg gctccgtcga tactatgtta tacgccaact ttgaaaacaa 9960
ctttgaaaaa gctgttttct ggtatttaag gttttagaat gcaaggaaca gtgaattgga 10020
gttcgtcttg ttataattag cttcttgggg tatctttaaa tactgtagaa aagaggaagg 10080
aaataataaa tggctaaaat gagaatatca ccggaattga aaaaactgat cgaaaaatac 10140
cgctgcgtaa aagatacgga aggaatgtct cctgctaagg tatataagct ggtgggagaa 10200
aatgaaaacc tatatttaaa aatgacggac agccggtata aagggaccac ctatgatgtg 10260
gaacgggaaa aggacatgat gctatggctg gaaggaaagc tgcctgttcc aaaggtcctg 10320
cactttgaac ggcatgatgg ctggagcaat ctgctcatga gtgaggccga tggcgtcctt 10380
tgctcggaag agtatgaaga tgaacaaagc cctgaaaaga ttatcgagct gtatgcggag 10440
tgcatcaggc tctttcactc catcgacata tcggattgtc cctatacgaa tagcttagac 10500
agccgcttag ccgaattgga ttacttactg aataacgatc tggccgatgt ggattgcgaa 10560
aactgggaag aagacactcc atttaaagat ccgcgcgagc tgtatgattt tttaaagacg 10620
gaaaagcccg aagaggaact tgtcttttcc cacggcgacc tgggagacag caacatcttt 10680
gtgaaagatg gcaaagtaag tggctttatt gatcttggga gaagcggcag ggcggacaag 10740
tggtatgaca ttgccttctg cgtccggtcg atcagggagg atatcgggga agaacagtat 10800
gtcgagctat tttttgactt actggggatc aagcctgatt gggagaaaat aaaatattat 10860
attttactgg atgaattgtt ttagtaccta gaatgcatga ccaaaatccc ttaacgtgag 10920
ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 10980
ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 11040
tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 11100
cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct 11160
gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 11220
gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 11280
tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 11340
ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 11400
gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 11460
ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 11520
tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 11580
ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 11640
gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 11700
acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt 11760
ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc 11820
tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg 11880
ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg 11940
catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac 12000
cgtcatcacc gaaacgcgcg aggcagggtg ccttgatgtg ggcgccggcg gtcgagtggc 12060
gacggcgcgg cttgtccgcg ccctggtaga ttgcctggcc gtaggccagc catttttgag 12120
cggccagcgg ccgcgatagg ccgacgcgaa gcggcggggc gtagggagcg cagcgaccga 12180
agggtaggcg ctttttgcag ctcttcggct gtgcgctggc cagacagtta tgcacaggcc 12240
aggcgggttt taagagtttt aataagtttt aaagagtttt aggcggaaaa atcgcctttt 12300
ttctctttta tatcagtcac ttacatgtgt gaccggttcc caatgtacgg ctttgggttc 12360
ccaatgtacg ggttccggtt cccaatgtac ggctttgggt tcccaatgta cgtgctatcc 12420
acaggaaaga gtccttttcg acctttttcc cctgctaggg caatttgccc tagcatctgc 12480
tccgtacatt aggaaccggc ggatgcttcg ccctcgatca ggttgcggta gcgcatgact 12540
aggatcgggc cagcctgccc cgcctcctcc ttcaaatcgt actccggcag gtcatttgac 12600
ccgatcagct tgcgcacggt gaaacagaac ttcttgaact ctccggcgct gccactgcgt 12660
tcgtagatcg tcttgaacaa ccatctggct tctgccttgc ctgcggcgcg gcgtgccagg 12720
cggtagagaa aacggccgat gccgggatcg atcaaaaagt aatcggggtg aaccgtcagc 12780
acgtccgggt tcttgccttc tgtgatctcg cggtacatcc aatcagctag ctcgatctcg 12840
atgtactccg gccgcccggt ttcgctcttt acgatcttgt agcggctaat caaggcttca 12900
ccctcggata ccgtcaccag gcggccgttc ttggccttct tcgtacgctg catggcaacg 12960
tgcgtggtgt ttaaccgaat gcaggtttct accaggtcgt ctttctgctt tccgccatcg 13020
gctcgccggc agaacttgag tacgtccgca acgtgtggac ggaacacgcg gccgggcttg 13080
tctcccttcc cttcccggta tcggttcatg gattcggtta gatgggaaac cgccatcagt 13140
accaggtcgt aatcccacac actggccatg ccggccggcc ctgcggaaac ctctacgtgc 13200
ccgtctggaa gctcgtagcg gatcacctcg ccagctcgtc ggtcacgctt cgacagacgg 13260
aaaacggcca cgtccatgat gctgcgacta tcgcgggtgc ccacgtcata gagcatcgga 13320
acgaaaaaat ctggttgctc gtcgcccttg ggcggcttcc taatcgacgg cgcaccggct 13380
gccggcggtt gccgggattc tttgcggatt cgatcagcgg ccgcttgcca cgattcaccg 13440
gggcgtgctt ctgcctcgat gcgttgccgc tgggcggcct gcgcggcctt caacttctcc 13500
accaggtcat cacccagcgc cgcgccgatt tgtaccgggc cggatggttt gcgaccgtca 13560
cgccgattcc tcgggcttgg gggttccagt gccattgcag ggccggcaga caacccagcc 13620
gcttacgcct ggccaaccgc ccgttcctcc acacatgggg cattccacgg cgtcggtgcc 13680
tggttgttct tgattttcca tgccgcctcc tttagccgct aaaattcatc tactcattta 13740
ttcatttgct catttactct ggtagctgcg cgatgtattc agatagcagc tcggtaatgg 13800
tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg atcctccgcc ggcaactgaa 13860
agttgacccg cttcatggct ggcgtgtctg ccaggctggc caacgttgca gccttgctgc 13920
tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct tttgctcatt ttctctttac 13980
ctcattaact caaatgagtt ttgatttaat ttcagcggcc agcgcctgga cctcgcgggc 14040
agcgtcgccc tcgggttctg attcaagaac ggttgtgccg gcggcggcag tgcctgggta 14100
gctcacgcgc tgcgtgatac gggactcaag aatgggcagc tcgtacccgg ccagcgcctc 14160
ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc gacacgacaa aggccgcttg 14220
tagccttcca tccgtgacct caatgcgctg cttaaccagc tccaccaggt cggcggtggc 14280
ccatatgtcg taagggcttg gctgcaccgg aatcagcacg aagtcggctg ccttgatcgc 14340
ggacacagcc aagtccgccg cctggggcgc tccgtcgatc actacgaagt cgcgccggcc 14400
gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg ccgacaacgg ttagcggttg 14460
atcttcccgc acggccgccc aatcgcgggc actgccctgg ggatcggaat cgactaacag 14520
aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg gttgcgatgg tcgtcttgcc 14580
tgacccgcct ttctggttaa gtacagcgat aaccttcatg cgttcccctt gcgtatttgt 14640
ttatttactc atcgcatcat atacgcagcg accgcatgac gcaagctgtt ttactcaaat 14700
acacatcacc tttttagacg gcggcgctcg gtttcttcag cggccaagct ggccggccag 14760
gccgccagct tggcatcaga caaaccggcc aggatttcat gcagccgcac ggttgagacg 14820
tgcgcgggcg gctcgaacac gtacccggcc gcgatcatct ccgcctcgat ctcttcggta 14880
atgaaaaacg gttcgtcctg gccgtcctgg tgcggtttca tgcttgttcc tcttggcgtt 14940
cattctcggc ggccgccagg gcgtcggcct cggtcaatgc gtcctcacgg aaggcaccgc 15000
gccgcctggc ctcggtgggc gtcacttcct cgctgcgctc aagtgcgcgg tacagggtcg 15060
agcgatgcac gccaagcagt gcagccgcct ctttcacggt gcggccttcc tggtcgatca 15120
gctcgcgggc gtgcgcgatc tgtgccgggg tgagggtagg gcgggggcca aacttcacgc 15180
ctcgggcctt ggcggcctcg cgcccgctcc gggtgcggtc gatgattagg gaacgctcga 15240
actcggcaat gccggcgaac acggtcaaca ccatgcggcc ggccggcgtg gtggtgtcgg 15300
cccacggctc tgccaggcta cgcaggcccg cgccggcctc ctggatgcgc tcggcaatgt 15360
ccagtaggtc gcgggtgctg cgggccaggc ggtctagcct ggtcactgtc acaacgtcgc 15420
cagggcgtag gtggtcaagc atcctggcca gctccgggcg gtcgcgcctg gtgccggtga 15480
tcttctcgga aaacagcttg gtgcagccgg ccgcgtgcag ttcggcccgt tggttggtca 15540
agtcctggtc gtcggtgctg acgcgggcat agcccagcag gccagcggcg gcgctcttgt 15600
tcatggcgta atgtctccgg ttctagtcgc aagtattcta ctttatgcga ctaaaacacg 15660
cgacaagaaa acgccaggaa aagggcaggg cggcagcctg tcgcgtaact taggacttgt 15720
gcgacatgtc gttttcagaa gacggctgca ctgaacgtca gaagccgact gcactatagc 15780
agcggagggg ttggatcaaa gtactttgat cccgagggga accctgtggt tggcatgcac 15840
atacaaatgg acgaacggat aaaccttttc acgccctttt aaatatccgt tattctaa 15898
<210> 5
<211> 96
<212> DNA
<213> Artificial sequence
<400> 5
gctggacggc cacgacctga gttttagagc tagaaatagc aagttaaaat aaggctagtc 60
cgttatcaac ttgaaaaagt ggcaccgagt cggtgc 96
<210> 6
<211> 20
<212> RNA
<213> Artificial sequence
<400> 6
gcuggacggc cacgaccuga 20

Claims (10)

1.一种蛋白质,为如下(a1)或(a2)或(a3)或(a4):
(a1)序列表中序列1所示的蛋白质;
(a2)在(a1)所述蛋白质的N端或/和C端连接标签得到的融合蛋白;
(a3)将(a1)经过一个或几个氨基酸残基的取代和/或缺失和/或添加得到的与植物发育相关的蛋白质;
(a4)来源于水稻且与(a1)具有98%以上同一性且与植物发育相关的蛋白质。
2.编码权利要求1所述蛋白质的核酸分子。
3.如权利要求2所述的核酸分子,其特征在于:所述核酸分子为如下(b1)或(b2)或(b3)或(b4):
(b1)编码区如序列表中序列2所示的DNA分子;
(b2)序列表中序列3所示的DNA分子;
(b3)来源于水稻且与(b1)或(b2)具有95%以上同一性且编码所述蛋白质的DNA分子;
(b4)在严格条件下与(b1)或(b2)限定的核苷酸序列杂交且编码所述蛋白质的DNA分子。
4.含有权利要求2或3所述核酸分子的表达盒、重组载体或重组微生物。
5.权利要求1所述蛋白质的应用,为如下(c1)或(c2):
(c1)调控植物的株高性状;
(c2)调控植物的叶夹角性状。
6.权利要求2或3所述核酸分子的应用,为如下(d1)或(d2):
(d1)培育株高性状改变的转基因植物;
(d2)培育叶夹角性状改变的转基因植物。
7.抑制权利要求1所述蛋白质的物质在植物育种的应用,所述育种的目标为如下(e1)或(e2):
(e1)培育株高降低的植物;
(e2)培育叶夹角变小的植物。
8.抑制权利要求2或3所述核酸分子的物质的应用,为如下(f1)或(f2):
(f1)培育株高降低的转基因植物;
(f2)培育叶夹角变小的转基因植物。
9.一种制备转基因植物的方法,包括如下步骤:在受体植物中导入抑制权利要求2或3所述核酸分子表达的物质,得到转基因植物;与所述受体植物相比,所述转基因植株株高降低和/或叶夹角变小。
10.一种植物育种方法,包括如下步骤:降低目的植物中权利要求1所述蛋白质的含量和/或活性,从而使目的植物株高降低和/或叶夹角变小。
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