專利名稱：：薏苡醇溶蛋白α-coixin基因、其編碼蛋白以及應用的制作方法
技術領域：
：本發明涉及薏苡(Coixlacryma-jobiL.)中醇溶蛋白a-coixin基因，其編碼蛋白以及應用。
背景技術：
：薏苡(Coix)屬禾本科薏苡屬，和玉米關系較近，同屬Maydeae族。薏苡在我國有悠久的栽培歷史，長期作為中藥材，而非糧食作物使用。我國薏苡種植面積在藥用作物中最大，約占藥材種植總面積的五分之一至四分之一，分布也最廣，幾乎全國各地都有栽培。近年來發現薏苡種子中含有結構極為獨特的薏苡仁酯，具有防治癌癥的作用，這也使得薏苡的研究引起了人們關注。薏苡種子除了含有薏苡仁酯外，其蛋白含量也很高，這個性狀引起了育種學家的廣泛關注。薏苡種子的蛋白質含量高達18.7%，為所有禾谷類種子最高。相比之下，水稻、玉米和小麥的種子中的蛋白含量都僅在10%左右。提高種子蛋白含量一直是作物育種的一個重要內容，也是近年來育種的熱點之一。薏苡種子的高蛋白含量與儲藏蛋白基因的高表達和積累有關。克隆和分析薏苡種子儲藏蛋白基因是研究薏苡高蛋白含量的一個關鍵，也將為玉米等禾本科作物的種子蛋白含量的遺傳改造提供新基因。因此，本研究的主要目的就是克隆和分析薏苡種子中主要儲藏蛋白基因。和玉米相比，薏苡的儲藏蛋白研究還很落后。玉米最主要的儲藏蛋白是a醇溶蛋白(a—zein)，由一個很大的基因家族所編碼，分為19kDa和22kDa兩個大類。其中22kDa醇溶蛋白研究的更為清楚，在玉米自交系B73中，共有16個基因，其中15個基因成簇分布在一個染色體位點上，被稱為22kDa醇溶蛋白基因簇(SongandMessing,2003，PNAS)。薏苡中也有與22kDa醇溶蛋白同源的儲藏蛋白，被稱為22kDa薏苡蛋白(22kDacoixin)，是薏苡種子中的主要儲藏蛋白。已有數據顯示，22kDa薏苡蛋白是個基因家族，而且像玉米中的醇溶蛋白那樣成簇分布，但其具體的組成和結構還不清楚。22kDa薏苡蛋白作為薏苡種子的主要儲藏蛋白，是研究薏苡儲藏蛋白基因的重點。
發明內容本發明的目的之一在于提供薏苡醇溶蛋白a-coixin基因17個成員的基因的編碼區序列。本發明的目的之二在于提供該基因中的10個成員基因編碼的氨基酸序列。本發明的目的之三在于提供該基因在改善植物蛋白質品質中的應用。為達到上述目的，本發明采用如下技術方案-一種薏苡醇溶蛋白a-coixin基因，其特征在于該基因具有下列核苷酸序列之一(1).具有SEQN01一18所示的DNA序列；(2).與序列表中序列1-18限定的核酸序列具有95%以上的同源性，且編碼相同功能蛋白質的DNA序列。上述的薏苡醇溶蛋白a-cobcin基因的編碼蛋白，其特征在于該編碼蛋白具有序列表中序列19-28的氨基酸殘基序列或者是將序列19-28的氨基酸殘基序列經過一個或幾個氨基酸殘基的取代、缺失或添加且具有與序列19-28的氨基酸殘基序列相同活性的由序列19-28衍生的蛋白質。上述的薏苡醇溶蛋白a-coixin基因在在改善植物蛋白質品質中的應用。本發明中發現有些薏苡醇溶蛋白中賴氨酸含量較高，幾乎是玉米醇溶蛋白中賴氨酸含量的2.5倍，在色氨酸含量上也明顯高于玉米醇溶蛋白。我們可以通過提高上述薏苡醇溶蛋白基因的表達量，來改善薏苡種子蛋白的品質，從而達到培育出優質蛋白薏苡的目的。同時也可以利用基因工程的手段，將上述基因導入到玉米胚乳中，來改善玉米種子蛋白的品質，達到培育優質玉米(高賴氨酸玉米)的目的。圖1顯示了a-coixin基因家族17個成員在兩個BAC上的分布結果。圖2顯示了a-coixin基因家族表達的11個成員在胚乳發育期(受粉后15-30天)中的表達數據具體實施例方式下面結合具體實施例，進一步闡述本發明。應理解，這些實例僅用于說明本發明而不用于限制本發明的范圍。下列實施例中未注明具體實驗條件的實驗方法，通常按照常規條件，分子克隆(MolecularCloning:ALaboratoryManual，3rded.)或植物分子生物學一實驗手冊(PlantMolecularBiology—ALaboratoryManual,MelodyS.Clark編，Springer-verlagBerlinHeidelberg,1997)中所述條件，或按照制造廠商所建議的條件。實施例一薏苡醇溶蛋白ct-coixin基因的克隆方法1.陽性BAC克隆的獲得方法根據NCBI得到的co/x/"(gi:296507)的序列，我們設計了兩對引物coixin/Pl、P2(5，ttgtgctccttgctctttcag3';5，cagataggcagcagggtttg3，)，通過PCR篩選體系從薏苡BAC文庫中分別篩選含coixin的BAC克隆。通過Southern雜交分析選定2個測序BAC克隆。大通量提取鳥槍文庫的質粒DNA，在MegaBACE4500上進行序列的測定。每個鳥槍質粒利用M13正反向引物，進行雙向測序。以上述兩個BAC克隆大小分別為120k和140k計算，以每個測序反應得到可用序列為500bp，得到可用序列的成功率以75%來計算，按10倍覆蓋的要求，共測序3200個鳥槍文庫質粒，即32塊％孔板。在RedHatLinux平臺的并聯計算機群中，使用Phred/Phrap/Consed程序拼接所測序列。構建亞克隆并測序以填補出現在序列之中的缺口。實施例二對兩個BAC的基因預測及同源比對利用基因預測程序FGENESH(http:〃www.softberry.com)預測序列中可能的基因(模式單子葉植物即monocot)。預測結果在(美國國立生物技術信息中心)即NCBI的GenBank中利用BLASTN及BLASTP(http:〃www.ncbi.nlm.nih.gov/BLAST/)程序進行同源比對分析，發現了17個a-coixin基因家族成員。參見圖1所示。實施例三通過使用反轉錄產物隨機測序的方法對a-coixin基因家族成員的表達情況及表達量進行研究。利用coixin基因家族中可擴增各個成員基因的通用引物通過RT-PCR擴增該家族成員。為了確保PCR擴增時不改變成員基因間的比例，PCR的循環數嚴格控制在線性擴增階段。隨后，PCR產物克隆入pMD18-T載體并隨即挑取克隆進行測序。由于薏苡a-醇溶蛋白基因中沒有內含子的存在，我們將測序的結果分別與每個基因的序列進行比對，從而確定它們的表達情況及表達成員mRNA的相對比例。結果如表1和圖2所顯示，其中有ll個成員基因是表達的，其中coixin-09的表達量最高占到總表達量的18%，其次是coixin-08和coixin-17分別占總量的14%，而coixin-ll和coixin-12只占了0.5%。由此看出，各表達成員的相對表達量是不同的，因此它們對薏苡種子中醇溶蛋白的含量高低的貢獻度也是不一樣的。表l:22kDcoixin基因家族各個成員的基本情況<table>tableseeoriginaldocumentpage6</column></row><table>實施例四表達成員基因翻譯后蛋白中氨基酸的組成與含量分析玉米是一種重要的農作物，在它的醇溶蛋白(zein)氨基酸的組成上，有兩種必需氨基酸嚴重缺乏賴氨酸(Lys)只占其蛋白的0.376%(1/266)而色氨酸(Tip)則更是缺失。通過對表達的a-coixin基因家族中成員基因翻譯后蛋白質中氨基酸成分及含量的比較研究，我們發現在這兩個重要必需氨基酸(Lys,Trp)的含量上有些coixin成員的含量較高。如coixin-02中有2個賴氨酸(Lys)占其蛋白總氨基酸的0.93%(2/215)，幾乎是玉米醇溶蛋白中該氨基酸含量(0.376%)的2.5倍。在色氨酸(Tip)的組成上，其中有6個coixin成員基因中都含有1個色氨酸(Trp)，而玉米醇溶蛋白(zein)中是缺失的，參見表2。這些重要的發現對于改造種子中蛋白質的品質有重要指導意義，如我們可以通過基因工程的手段來提高coixin-02的表達含量(注目前其含量只占總量的2%，有很大的上升空間)達到培育出高賴氨酸的優質蛋白薏該，還可以通過轉基因等技術將含有高賴氨酸(Lys)含量的和色氨酸(Tip)的coixin成員基因轉入玉米胚乳中，來改善其蛋白品質等。表2:表達的22kDcoixin家族成員基因的氨基酸組成及氨基酸的含量(c代表coixin)<table>tableseeoriginaldocumentpage7</column></row><table>〈120〉薏苡醇溶蛋白a-coixin基因、其編碼蛋白以及應用<160>28<210>1<211>507<212>DNA<213>禾本科薏苡屬(Co〖x/acT7脂:/oW丄.)<400>1cgctcagtcaactagccatgacaaaccctgccacctatctacaaccacaacttttttctc60aaacccactagccgcggccaatgtcgttgcatacctgcaacaatagcacctacaacagat120cctaccaacgctcagtcaactagctattgcaaaccctgccgcgcgcctatctacaacagc180aactatttccattcaacccattagccgcgaccaatgctgttgcatacctgcaacaacaac240tacagcaaatcctaccaggcgctcagccaactagccgtggcgetacactgcctcctacatg300caacaacaacttcttcgttttaaccaactggctatggcgaacaccgctgcaacaacatta360gctactacaagttaaaccactgccaatgggttggttgtcgccttcctgcagcagcagcac420gaactactactgtcattcaaccaaatgtctttgatgaaccctgccttatcatggttgcaa480cccatcattggaggtgtcatcctctag507<210>2<211>1797<212>DNA<213>未本科薏苡屬(Co〖;c/aajmflf:/o6/Z(.)<400>2misc_feature171..175/note="endospermaboxmiscfeature200..209/note="opaque-2boxmisc—feature333..338/note=，，CATCbox"TATA_signal402..409/codon—start=l/product=，，alpha-coixin'，gcatatgttcatcaattgatg犯taacttctcacctaaca犯cgtgcatggcttaacaat60gagtacaatactaaaccaaagtcatttgatccaataagcaagtcaaaacattacgtactc120atcatgtttataaagtggaacattcatgatgtggctaaggctaaaacatatgtaaaggtg180aagtgatcatgcatgtcattcctcatgaattggaagaattccaatagaaagcgacacttt240ttcatgtatgtagtggaaactatctttccaacacaaaccatacaatccaataaaactgag300aactaaatgacaaaatcgactaggagccatgtcatctctagcttatctgcttcttggcaa360aatctaaaatttgaacacacatacatgagcttatagacttgtataaatagggcccaaatc420aatagttaattcatcgcccataacagtgagatcaattaggaacataccaagtgaaacaca480ctagcagcatcttggcaagaatggctaccaagatatttgccctccttgtgctccttgctc540tttcagcgagcgctacaactgcggtcattattccgcagtgctcacttgctcctactgttg600ctattatttcaaagttcctcccacttgtttctgctatagggttcgaacacccagctgtgc660gggcctacaggctacaacaggcgcttgcggcaagcatcttgc犯caaccacttgcccaat720tgcaacaacagtcctcggctcatctaaccatacagacaatcgcagcacaacagcaacaac780agttcctgccagcactcagtcaactagcagccacgaacccagctgcatacttgcaacaac840agctgcttgcttccaacccacttgttgtggcaaacgctattgcataccagcaacaacaac900agctgcaacagttcctgccagcgctcagtcaactagccgtggcaaaccctgctgcctatc960tgcaaccacaactatttccttccaacccactagccgcggccaatgctgctgcatacctgc1020aacaacagcagctacaacagatcctaccggcgctcagtcaactagccgtggcgaacccta1080actcctacttgcaacaacaacaacttcttccattcaaccaagtggctgtggcgaacaacg1140ctgtatactagcaacaacatcagctgctgc肪gttaacccattggctgccgccttcctac1200agcagcagcaacaactactactgccattcaaccagatgtctttgatgaaccctgccttgt1260cgtggcaacaacccatcgtcggaggtgtcggcttctagattaaatatgggttgtacttga1320taaaaaagctctcatgccggcgtgtgcaacttctaagaaataatcaatatactgattgag1380atttatcagagtctatttcttttctgtgttttccgtattaaaaattaaaaacattgtatc1440ataactaaaggcacatggttggttaattcgtagacaataccacatcacttcatcaatcca1500aaattgtggcagcccaagctcttcaacattctcatttgggtgggtgggtgtgtttataag1560atagacgtaattgtacatgcttgcagttgagtaattgcatacgtagtgtattgtttcaag1620aaagtgatcggcacatataccttgactatggccttgatcttaggtgcacgtataccaagc1680aaaacaaaatctatgtttatgggtaaactgatatatgcatccaaagttgtttgcggatag1740tttgaccatgtaaatattgtttttagtaaaatatctggtgaatatgcttttaactat1797<210>3<211〉1800<212>DNA<213>禾本科薏攻屬(Cofx/acr^ma-^&i.)<400>3misc—feature171..175/note="endospei:mabox"misc一feature200..209/note="opaque-2box"misc—feature335..340/note="CATCbox"TATA—signal402..409CDS501..1301/codon—start=1/pr〇duct=，'alpha-coixingcatttgttcatcaattgatgtataacttctcacctatcaaacgtgcatggcttaacaac60gagtacaatactaaaacaaagtcatttgatccaatgagcaagtcaaaacattacatactc120agcatgtttataaagtggaacctttatgatgtggctaaggctaaaacgtatgtaaaggtg180aagtgatcatgcatgtcattcctxatgaattagaagaattccgLatagaaagcgacacttt240ttttcttgtatgtagtggaaactatctttccaacacaaaccatacaatccaacgaaactg300agaactaaatgacaaaattgactaggagacatgtcatctctagcttaixtgcttgttggc360aaaatctaaaatttgaacacacatacatgagcttagacttgtataaatagggcccaaatc420aatagttaatccatcgcccataacactgagettcaattaggaacataccaagtgaaacaca480ctagcagcatcttggcaagaatggctaccaagatatttgccctccttgtgctccttgctc540tttcagcgagcgctacaactgcggtcattattccgcagtgctcacttgctcctactgctg600ctattattcagcggttcctcccacctgtttctgctatagggttcgaacacccagctgtgc660aggcctacaggctacaacaggcgcttgcggcaaccatcttacaacaaccacttgcccaat"720tgcaacaacggtcctcggctcatctaaccatacagacaatcgcagcacaacaacaacaac780aacagttcctggcatcactcagtcaactagcagccgcgaacccagctgcatacttgcaac840aacagctgctggcatxcaacccacttgctgtggcgaacgctattgaataccagcaacaac900aacagctgcaacagttcctgccagcgctcagtcaactagtcgtggcaaaccctgctgcct960atctgcaatcacaattgtttccatccaacccactagtcgtgaccaatgccgctgcatacc1020tgcagcaacagcagctacaacagatcctaccagcactcagtcaactagccgtggcgaacc1080ctaetctcctacttgcaacaacaacaacttcttccattcaaccaagtggctgtggcgaaca1140acgctgtataccagcaacaatatcagctgctgc犯gttaacccattggttgccaccttcc1200tacagcagcagcaacggcaactactgccattcaaccaaatgtctttgatgaaccctgcct1260tgtcgtggcagcaacccatcgttggaggtgtcggcttctagattaaatatgagttgtact1320tgataaaaaagctctcatgccggcttgtgcaacttcctagaaataatcaatatattgatt1380gagatttatcagagtctatttcttttctgtgttttccgtatgaaaaattaaaaatattgt1440atcataactaaaggcacatggttggttaattcgtagacaeitaccacattacttcatcaat1500ccaaaattgtggcagcccaagctcttcaacattctcatttgggtgggtgggtgtgtttat1560aagatagacgtgattgtacatgcttgcagttgagtaattgcataggtagtgtattgtttc1620aagaaagtgattgacacatatgctttgaccatggcccttcatcttaggtgcacctatacc1680gagcaaaacaaaatctatgtttatgggtaaaccgatatatgcattcaaagttgttgtgga1740tagtatgaccttgtaaatattgttttaagtaaaatatctggtgattatgcttttaactat1800<210>4<211>801<212>DNA<213>禾本科薏苡屬(Coi'x/ac，a-/o6".)<400>4atggctaccaagatatttgccctccttgtgctccttgctctttcagcgagcgctacaact60gcggtcattattccgcagtgctcacttgctcctactgctgctattattcagcggttcctc120ccacctgtttctgctatagggttcgaacacccagctgtgcagtcct3C3ggct3c犯cag180gcgcttgcggcaagcatcttacaacaaccacttgcccaattgcaacaacagtcctcggct240C3tct33ct3t3C3gaca3tcgcagccc犯caacaacaacaacagttcctgccatcactc300agtcaactagcagccgtgaacccagctgcatacttgcaacaacagctgctggcatccaac360ccacttgctgtgccgaacgctattgaataccagc犯c犯c犯cagctgcsacagttcctg420ccagcgctcagtcaactagccgtggcaaaccctgctgcctatctgcaatcacaactgttt480ccatccaaccaactagtcgcgaccaattccgctgcatacctgcagcaacagcagctaca^540cagatcctaccggcgctcagtcaactagccgtggcg犯ccCt￡L￡LCtCCt￡LCttgC￡LaC￡L￡L600caacaacttcttccattcaaccaagtggctgtggcga織acgctgtatactagc肌caa660catcagctgctgcaagttaacccattggttgccgccttcctacagcagcagc犯cggc犯720ctactgccattcaaccaaatgtctttgatg犯ccctgccttgtcgtggcagcaacccatc780gttggaggtgtcggcttctag801<210>5<211>1798<212>DNA<213>禾本科薏苡屬(Co/"ac，一6i丄.)<400>5misc一feature172/note=misc_feature201/note=misc—feature335/note=12、.176"endospermabox..210"opaque-2box"..340"CATCbox"TATA—signal402..409CDS501..1298/codonstart=l/product-"alpha—coixinagcatttgttcatcaattgatgaataacttctcacctaacaaacgtgcatggcttaacaacgagtacaatacaaaaacei3agtcatttgatcceiatgeigcaagtcaaei3tattacstactcatcatgtttataaagtggaacctttatgatgtggctaaggctaaaatgtatgtaaaggtgaagtgatcatgcacgtcattcctcatgaattagaagaattccaatagaaagtgacacttttttcttgtatgtagtggaaactatctttccaacacaaaccatacaatccaatgaaactgagaactaaatgacaaaatct3Ctaggagccatgtcatctctagcttatctgcttcttggc肌aatctgigiEi￡itttgaaca_cacatacatg3gcttagax;ttgtataaatagggcccsaatcaatagttaatccatcgcccataacactgag3tcaattaggaacatacc肪gtgaaacaicactagcagcatgttggcaagaatggctaccaagatatttgccctccttgtgctccttgctctttcagggagcgctacaactgcggtcattattccgcagtgctcacttgctcctactgctgctattattccgcggttcctcgcacctgtttctgctatagggttcgaacacccagctgtgcaggcctacaggctacaacaggcgcttgcggcaagcatcttacaacaaccacttgcccaattgcaacaagagtcctcggctcatctagccatacagacaatcgcagcacaacaac肪caacagttcctgccatcactcagtcaactagcagccgcgaacccagctgcatacttgcaacaacagctgcttgc3ttcaacccacttgctgtggcgaacgctattgcataccagcaacaacaacagctgcaacagttcctgccagcgctcagtcaactagccgtggcaaaccctgctgcctatctgcaatcacaactgtttccatccaacccactagtcacggccaatgtcgctgcatacctgcagcaacagcagctacaeicagatcctaccggcgctcagtcaactagccgtggcgaaccctaactcctacttgcaacaacaacaacttcttccattcaaccaagtggctgtggcgaacaacgctgtataccagcaacaaaatcagttgctgcaagttaacccattggctgccgccttccaacagcagcagcaacggcaactactgccattcaaccagatgtctttgatgaaccctgccttgtcgtggcagcaacccatcgttggaggtgtcggcttctagattaaatatgagttgtacttgataaaaaaagctctcatgccggcgtgtgcaacttccteigaaataeitcaMatattgattgagatttatgagagtctatttgttttctgtgttttccgtatgaaaaattaaaaacattgtatcataact3朋ggcacatggttggttaattcttagacaataccacattacttcatcaatccaaaattatggcagccc朋gctcttcaacattcttatttgggtgggtgggtgtgtttataa600360420480540600660720780840■9600001560gatsgacgtgtctgtacstgcttgc8gttgagtaattgcataggteigtgtattgtttcaa1620gaaagtgattgacacatatgctttgaccatggcccttcatcttaggtgcatgtataccga1680gcaaaacaaaatctatgttt3tgggt犯accgatatatgcactca^gttgttgtggata1740gtatgaccatgtaaatattgtttt肌gtaaaatatctggtgattatgcttttaactat1798<210>6<211>253<212>DNA<213>禾本科薏攻屬(Co/x/"cr;w"-yoW丄.)<400>6atgactaccaagatatttgccctccttgtgctccttgc1xtttcagcgagcgctscaact60gcggtcattattccgcagtgctcacttgctcctactgctgctattattccgcggttcctc120ccacctgtttctgctatagggttcgaaaacccagctgtgcaggcctacaggctacagtag180gcgcttgcggcaagcatcttacaacaaccacttgctcaattgC￡L3CaaC3gtcctcggct240catctaaccatac253<210>7<211〉549<212>DNA<213>禾本禾斗薏該屬(Co/x/"ajma-yoW丄.)<400>7atacagacaatcgcagcacaacaaca^caacagttcctgccatcactcagtcaactagca60gccgcgaacccagctgcatacttgcaacaacagctgcttgC3tCC犯CCCacttgctgcg120gcaaacgctattgcataccaccascaacaacagctgcaacagttcctgccagcgctcagt180caactagccgtggcaaaacctgctgcctatctgcaatcacaactgtttccEitccaaccca240ctagttgcggccaatgtcgctgcatacccgcagcaacagcgatcctaccg300gtgctcagtcaactagccgtggtgaaccctaactcctacttgcaaceieicELacettcttxtt360ccattcaaccaagtggctgtggcgaacaacgctgtatact3gcaac犯catcagctgctg420caagttaacccattggctgtcgccttcctacagcagcagcaacggcaactcctgccattc480aaccagatgtctttgatgaaccctgccttgtcgtggcagcaacccatcgttggaggtgtc540ggcttctag549<210>8<211>1798<212>DNA<213>禾本科薏苡屬(Coix/ac7附a:/o6/厶)<400>8misc一feature176..180/note=''endospermabox"misc—feature205…214/note="opaque-2box"misc—feature334…339/note="CATCbox"TATA_signal402…409CDS501…1298/codon—start=l/product="alpha-coixin"tgatagcatttgttcatcaattgatgaataacttctcacctaacaaacgtgcatggctta3C3glCgaCt3C33t3Ct朋33C肪3gtC3tttg3tCC33tg3gC33gtC3333C3tt3C3tactcatcatgtttataaagtggaacctttatgatgtggctaaggctaaaacgtatgtaaaggtgaagtgatcatgceitgtcattcctceitgaattagaagaattccaatagaaagcgacacttttttcttgtatgtagtggaaacaatctttccaacacaaaccaaacaatccaatgaaactgagaactaaatgacaaaatcgactaggagccatgtcatagcttatctgcttcttggcaaaatctaaaatttgaaca_ca^catacatgagcttageicttgtataaatagggcccaaatcEiatagttaatccatcgcccataacactgagatcaattaggaacataccaagtgaaacacactagcagcatcttggcaagaatggctaccaagatatttgccctccttgtgctccttgctctttcagcgagtgctacaactgcggtcattattccgcagtgctcacttgctcctactgctg600360420480540600ctattattccgcggttcctcccacctgtttctgctgtagggttcgaacacccagctgtgc660aggcctacaggctacaacaggcgcttgcggcaagcatcttacaacaaccacttgaccaat720tgc犯caacagtcctcggctC3tctaaccatacagacaatcgcagcacaacaac犯caac780agttcctgccatcactcagtcaactagcagccgcgaacccagctgcatacttgcaacaac840agctgcttgcattcaacccacttgctgtggcgaacgctattgcataccaccaacaacaac900agctgcaacagttcctgccagcgctcagtcaactagccgtggcaaaccctgctacctatc%0tgcaatcacaactgtttccatccaacccactagtcgcggccaatgtcgctgcatacctgc1020agcaacagcagctacaacagatccttccggcgctcagtcaactagccgtggcgaacccta1080actcctacttgcaacaacaacaacttcttccattcaaccaagtggctgtggcgaacaacg1140ctgtataccagcaacaacatcagctgctgcaagttaacccattggctgccgccttcctac1200agcagcagC3acggc朋ctactgccsttcaaccagatgtctttgatgaaccctgccttgt1260cgtggcagcaacccalcgttggaggtgtcggcttctagattaaatatgagttgteicatga1320taaaaaaagctctcatgccggcatgtgcaacttcctag犯at犯tcaatatattgattga1380gatttatgsgagtctatttcttttgtgttttccgtatg朋a朋tt朋a朋C3ttgt3tca1440taacta犯ggcacatggttggttaattcgtagacaataccacattacttcatcaatccaa1500aattatggcagcccaagctcttcaacattctcatttgggtgggtgggtgtgtttataaga1560tagacgtgtctgtacatgcttgcagttgagt犯ttgcataggtagtgtattgtttcaaga1620aagtgattgacacatatgctttgacca/tggcccttcatcttaggtgcatgtataccgagc1680aaaacaaaatctatgtttatgggtaaaccgatatatgcattcaaagttgttgtggatagt1740atgaccatgtaaatattgttttaagtaaaatatctggtgattatgcttttaactat犯1798<210〉9<211>1798<212>DNA<213>禾本禾斗薏攻屬(O^c/acrj/ma-yoWZ.)<400>9ruisc—feature174..178/note="endospermaboxmiscfeature203..212/note="opaque-2box"misc_feature337..342/note="CATCbox"TATA—signal404..411CDS501..1298/codon—start=1/product="alpha-coixinatagcatttgttcatcaattgatg犯taacttctcccctaacaaacgtgcatggcttaac60aacgagtacaat3ctaaeLacasagtca/tttgatcc犯tgagcaagtca^aacattaceits120ctcatcatgtttataaagtggaacctttatgatgtggctaaggctaaaacgtatgt犯ag180gtgaagtgatcatgcatgtcattcctcatgaattagaagaattccaatagaaagcgacac240ttttttcttgtatgtagtggaaacaatctttccaacacaaaccatacaatccaatg犯ac300tgagaactaaatgacaaaatcgactaggagccatgtcatctctagcttatctgcttcttg360gc犯犯tct3朋3tttg3ac3C3C3t3C3tg3gctt3gscttgtst3犯t3gggccc3朋420tcaatagttaatccatcacccataacactgagatcaattaggaacataccaagtgBcaca480ctagcagcatcttggcaagaatggctaccaagatatttgccctccttgtgctccttgctc540tttcagcgagcgctacaactgcggtcattattccacagtgctcacttgctcctactgctg600ctattattccgcggttcctcccacctgtttctgctatagggttcgaacacccagctgtgc660aggcctacaggctacaacaggcgcttgcggcaagcatcttacaacaaccacttgcccaat720tgcaacaacagtcctcggctcatctaaccatacagacaMcgcagcac幼caacaacaac780agttcctgccatcactcagtcaactagcagccgcgaacccaactgcatacttgcaacaac840agctgcttgcatccaacccacttgctgtggcgaacgctattgcataccagcaccaacaac900agctgcaacagttactgccagcgctcagtcaactagccgtggcaaaccctgctgcctatc%0tgcaatcacaactgtttccatccaacccactagtcgcggccaatgccgctgcatacctgc1020agcaacagcagctacaacagatcctaccggtgctaagtcaactagccgtggcggscccta1080actcctacttgcaacaacaacaacttcttccattcaaccaagtggctgtggcgaac犯cg1140ctgtatacgagcaacaacatcagctgctgcaagttaacccattggctgccgccttcctac1200agcagcagcaacggcaactacttccattcaaccagatgtctttgatgaaccctgccttgt1260cgtggcagcaacccatcgttggaggtgtcggcttctaaattaaatatgagttgtacttga1320tg犯33a3gctctcatgccggcgtgtgc犯cttcctag犯at3atc3atat3tag3ttga1380gatttatcagagtctatttcttctctgtgttttcgtatgaaaaattaaaaac3ttgtatc1440ataactaaaggcacatggttggttaattcgtagaceiataccacattacttcataaatcca1500aaattatggcagcccaagctcttcaacattctcatttgggtgggtgggtgtgtttat犯g1560at已gacgtgattgtacatgcttgcagttgtgtaMtgcacaggtagtgtattgtttcaag1620aaagtgattgacacatatgctttgaccatggcccttcatcttaggtgcacgtat8CCg3g1680caaaac犯aatctatgtttatgggt犯accga/tatatgcattc犯agttgttgtggst3g1740tatgaccatgtaaatattgtatctggtgattatgcttttaactataactc犯actgtg1798<210〉10<211>1795<212>DNA<213>禾本禾斗薏攻屬(Co/jc/"c77附":/o6f厶)<400>10raisefeature172..176/note="endospe:i:mabox"iuiscfeature201..210/note="opaque-2box"miscfeature335..340/note="CATCbox"TATA—signal402..409CDS5〇1..1295/codonstart=l/product二"alpha-coixin"agcatttgttcatcaattgatgaataacttctcacct犯caaacgtgcatggcttagcaa60cgagtacaatact犯犯c犯agtcatttgatcc犯tgagcaagtca犯aca<ttac￡Ltact120catcatgtttataaagtggaacctttatgatgtggctaaggctaaaatatatgt犯aggt180gaagtgatcatgcatgtcattcctcatgaattagaagaattccaatagaaagegacaett240ttttcttgtatgtagtggaaactstctttccaacacaaaccstacaatcc犯tg已犯ctg300已gaactaaatgacaa犯tcgactaggageeatgtcatctctagcttatctgcttcttggc360a犯gitctaaa3tttgaacacacatacatgagcttagacttgtata犯tagggccccaatc420aatagttaatcc3tcgcccataacactgeigaacaattagggtg￡Laa<c3ca_480ctagcagcatcttggcaagaatggctaccaagatatttgccctccttgtgctccttgctc540tttcagcgagcgctacaactgcggtcattattccgcagtgctcacttgctcctactgctg600ctattattccgcggttcctcccacatgtttctgctatagggttCg￡L￡LC￡lCccagcagtac660aggcctacaggctac已acaggcgcttgcggcaagcatcttcttgcccaM720tgcaacaacagtcctcggctC￡Ltct￡L￡lCCatacagac犯tCgC￡LgCBCa￡Lc犯caBC3gt780tcctgccatcactcagtcaacaagcagccgtgaacccagctgcatacttgcaac^cagc840tgcttgcatccaacccactcgctgtgccgaacgctattgcataccagcaa■tgcaacagttcttgccagcgctcagtcaacttgccgtggcaaaccctgctgcctatctgc960aatcacaactgtttccatgcaacccactagtcgcggccaatgccgctgcatacctgcagc1020aacagcagctacaacagatcctaccggcgctaagtcaact已gccgtggcgaaccctaact1080cctacttgcaacaacaac犯cttcttccattcaaccaagtggctgtggcgaacaacgctg1140tatacgagcaacascatcagctgctgcaagttaacccattggctgccgccttcctacagc1200agcagcaacggcaactacttccattcaaccagatgtctttgatgaaccctgccttgtcgt1260ggcagcaacccatcgttggaggtgtcggctatatgagttgtacttgatg31320aaaaaagctctcatgccggcgtgtgcaacttccta>g￡iaMteigattgeiga1380tttatcagagtctatttcttctctgtgttttcgtatg犯aattgtatcat1440aactaaaggcacatggttggttaattcgtacattacttcataaatccaaa1500attatggcagcccaagctcttcaacattctcatttgggtgggtgggtgtg1560agacgtgattgtacatgcttgcagttgtgt犯ttgcax:aggtagtgtattgtttcaag犯1620agtgattgacacatatgctttgacceitggcccttcatcttaggtgcacgtataccgsgc31680犯eica犯atctatgtttatgggtaaaccgatatatgcattcaaagttgttgtgg￡Ltagt￡L1740tgaccatgta犯tattgtatctggtgattatgcttttaacactgt1795<210>11<211>797<212>DNA<213>禾本禾斗薏苡屬(CoZx/tfcoTno^/oWi.)<400〉11atggctaccaagatatttgccctccttgtgctccttgctctttcaacgagcgctacaaca60gcggtcattattccgcagtgctcacttgctcctactgctgctattattccgcggttcctc120ccacctgtttctgttatagggttcgagcacccagctgtgcaggcctacaggctacaacag180gcgcttgcggcaagcatcttacaacaaccacttgcccaattgcaacaacagtcctcggct240catctaaccatacaaac￡L￡itcgcaacacaacaacaacaacagttcctgccatcactcagt300caactagcagccacgaacccagctgcatacttgcaacaacagctgcttgcatccaaccca360cttgctgtggcaaacgctattgcataccagcaacaacaacaactgcaacagttcctgcta420gcgccagtcaactagccgtggcaaaccctgctgcctatctgcaatgacaactgtttccat480ccaacccactagtcgcggccaatgccgctgcatacctgcagcaacagcagctacaacaga540tcctaccggcgctcagtc犯ctagccgtggcgaaccct犯ctcctacttgc犯c犯caac600aacttcttccattgaacc犯gtggttgtggcgaacaacactgtataccagcaacaacatc660agctcctgcaagttaacccattggctgccgccttcctacaacagcagcaacaacaactac720tgccattcaaccagatgtatttgatgaaccctaccttgtcgtggcaacaacccatcattg780gaggtgtcggcttctag797<210>12<211〉1802<212>DNA<213>禾本禾斗薏苡屬(Co/x/ao7W"證/o6Z丄.)<400〉12misc一feature172..176/n〇te=，'endospei:maboxmisc—feature201..210/note="opaque-2box"misc—feature335..340/note="CATCbox"TATA—signal402..409agcatttgttcatcaattgatcaataacttctcacctaacaaacgtgcatggcttaacaa60cgagtacaatactaaaacaaagtcatttgatcctatgagcaagtc犯aacattacatact120catceLtgttt￡Lt￡La￡tgtggaacatttstgatgtggct犯ggct犯朋cgtatgt犯3ggt180gaagtgatcatgcatgtcattcctcatgaattagaagaattcc犯tag犯agcaacactt240ttttcttgtatgtagtggaaactatctttccaacacaaaccatacaatccaatgaaeictg300agaactaaatgacaaaatcgactaggagccatgtcatctctagcttatccgcttcttggc360aaaatctaacatttgaacacacatacatgagtttagacttgtataaatagggcccaaatc420aatagttaatccatcgcccataacactgagatcaattaggaacataccaagtgaaacaca480ctagcagcatcttggcaagaatggctacctagatatttgccctccttgtgctccttgctc540tttcagcgagcgctacaactgcggtcattattccacagtgctcacttgctcctactgctg600ctattattccgcggttcctcccacctgtttctgctgtaggggttcgaacacccagctgtg660caggcctacaggctacaacaggcgcttgcggcaagcatcttacaacaaccacttgcccaa720ttgcaacaacagtcctcggctcatctaaccatatagacaatcgcagcacaacaacaacaa780caacagttcctgccatcactcagtcaactagcagccacgaacccagctgcatacttgcaa840caacagctgcttgcatccaacccatttgatgtggcgaacgctattgcataccagcaacaa900caacagctgcaacagttcttgccagcgatcagtcaactagccgtggcaaaccctgctgcc960tatctgcaatcacaactgtttccatccaacccactagtcgcggccaatgccgctgcgtac1020ctgcagccacagcagctacaacagatcctaccggtgctcagtcaactagccatgatgaac1080cgtaactcctacttgcaacaacaacaacttcttccattcaaccaagtggctgtggcgaac1140aatgctgtataccagcaacaacatcagctgctgcaagttaacccattggatgccgccttc1200ctacagcagcagcaacaacaactactgccattcaaccagatgtctttgatgaaccctgcc1260ttgtcgtggcaacaacccatcattggaggtgtcgacttctagattaaatatgagttgtac1320ttgataaaaaaagctctcatgccagcgtgtgcaacttcctagaaataatcaatatattga1380ttgagatttatcagagtttatttcttttctgtgtttttcgtatgaaaaaataaaaacatt1440gtatcgtaactaaaggcacatggttaattcgtagacaataccacattacttcatcaatgc1500a犯attgtggcagcccaagctctacaacsttctcatttgtgtgggtgggtgtatttac犯1560gatagacgtgactgt3catgcttgcagttgagt犯ttgcataggt昭tgtattgtttc朋1620gaaagtgattgeicacatatgctttgactatggcccttcatcttaggtgcacgtataccga1680gc犯aacaaaatctatgtttatgggtaaagcgatatatgcatccaaagttgttgtggata1740gtatgaccatgtaaatattgtttttagtaaaatatctggtgattatgcttttaactataa1800ct1802<210>13<211>1940<212>DNA<213>禾本禾斗薏苡屬(Co/;c/acr;;wa:/oW丄.)<400>13miscfesture172，.176/note="endospermabox"miscfeature201.，210/note="opaque—2box"miscfeature335，，340/note="CATCbox"TATA一signal402.409CDS501，，911/codonstart=l/pr〇duct="alpha—coixinagcatttgttcatcaattgatgaat犯cttctcacctaacaaacatgcatggcctaacaa60tgagtataatactaaaacaaagtcatttgatccaatgagcaagtc犯aacattctgtact120cgtcatgtttat犯agtggaacatttatgatgtggctaaggctaaaacatgtgtaaaggt180gaagtgaccatgcatgtcattcctcatgaatcag肪gaattccaatagaatgcgacactt240ttttcttgtatgtagtggaaactatcttttcaacacaaaccatacaatccaatgaatctg300ataactaaatgacataatcgactaggagccatgtcatctctagcttatctgctttttggc360ataatctaaaatttcaacacacgtacatgagcttagacttgtataaatagggctcaaatc420aatagttaatccatcacccattgcactgagaggaattagaaacataccaagtgaaacaca480ctagcagcatcttggcaacaatggctaccaagatatttgtcctccttgtgctccttgctc540tttcagcgagcgctacaactgcggtcattattccacagtgctcacttgctcctagtgctg600ctattattccacagttcctcccaaccgtttctgctatagggttcgaacacccagctgtgc660aggcctacaggctacaacaggcgcttgcgtcaagcatcttacaacaaccacttatccaat720tgcaacaacagtcctcggctcatctaaccatacagaccatcgcaacacaacaacaacaac780agttcctgccagcgctcagtcaactagtagcagcgaacccagctgcatacttgcaacaac840agttgtttgcatccaacccactttctgtggcgaacgctgttgcataccagcaacaacaat900attcacaatagttcctgccagcgctcagtcaactggctgtggcaaaccctgttgcctatc960tgcaagcacaactgtttccatccaacccactagccgcggccaatgccgctgcatacctgc1020tacaacagcagctacaacagatcctaccagcgctcagtcaactagctgtcgcaaaccctg1080ccgcctatctgcaacaacatcttttttcatccaacgcactagccgcggccaatgcagctg1140catacctgcaacaacaacagctacaacagatcctaccagcactcaagtcaactagccatg1200gcgaaccctgcctcctacttgcaacsctaacttcttccatttaaccaactggctgtggcg1260朋c朋cgttgC3t3ccagc3acaacatcagttgctgc33gtt朋cccactggt3gtggct1320aacccattggctgccacctttctacagcagcagcaacaacaactaccgccattcaaccag1380atgtctttgatgacccctgccttgtcatggcagcaacccatcgttggacgtgtcttctag1440attaaateitgagttgtacttgataaaaaggctttcatgccgacatgtgcaacttcctaga1500aataatcaatatattgattgagatttattatagtctatttcttatttgtgttgttcatgt1560gaaaaattaaaaacatcgtatcataactaaaggceicatggttagttaattcatagataat1620accacattacttcatcaatccaaatatgtggaaacccaagatcttcaacgttctcatttg1680ggtgtgtgtgtttgtttataagatacatgtgactgtacatgctcgcagttgagtacttca1740taggtagtgtgttgtttcaagaaagtgattggcacatatgctttgaccatggtcgttcaa1800aataacacccccttttgcaacacatatatgtgttgataaaagtctatatatgtgttggtg1860aagtttataccaacacattaactatatgtcgctagaagtggcgttgcaagggtctattgc1920aacacttgtacatacgttgg1940<210>14<211>1939<212>DNA<213>禾本禾斗薏該屬(Coix/"aym^/oZ)/丄.)<400>14misc—feature172..176/note="end〇spermaboxmisc—feature201..210/n〇te="opaque-2box',misc—feature335..340/note="CATCbox"TATA—signal402..409CDS501..1439/codon—start=l/product-"alpha-coixingattagatgtaagtgttaatatagttatgtttacgttatgtataacattgtttctgatat60tctttattctgttgctgtatgtgtggacttcctgggtacacatatggtgagcctggtttt120gtttttaaaaccgggtgtgacaatttatgatgtggctaaggctaaaacatgtgtaaaggt180gaagtgaccatgcatgtcattcctcatgaatcagaagaattccaatagaatgcgacactt240ttttcttgtatgtagtggaaattatctttccaacacaaaccatacaatcgaatgaaactg300atcactaaatgacataatcgactaggagccatgtcatctctagcttatctgctttttggc360aaaeitctei犯atttgaacacEicatacatgeigctteigacttgtat犯a"tagggctc犯atc420aatagttaMccatcacccattgcactgagagcaattagaaacataccaagtgaaacaca480ctagcagcatcttggcaacaatggctaccaagatatttgtcctccttgtgctccttgctc540tttcagcgagcgctacatccgcggtcattattccgcagggctcacttgctcctactgctg600ctattattccgcagttcctcccaaccgtttctgctatagggttcgaacacccagctgtgc660aggcctacaggctacaacaggcgcttgcgtcaagcatcttacaacaaccacttatxcaat720tgcaacaacagtcctcggctcatctaaccatacagaccatcgcagcacaacaacaacaac780agttcctgccagcgctcagtcaactagtagcagcgaacccagctgcatacttggaacaac840agttgcttgcatcta^cccacttgctgtggcg犯cgctgttgcataccagcaac犯c犯t900attcacaacagttcctgccagcgctcagtcaactggctgtggcaaaccctgctgcctatc960tgcaagcacaactgtttccatccaacccactagctgcggccaatgccgctgcatacctgc1020犯c犯cggcagctacaacaggtcctaccagcgctcagtcaattagttgtcgcaaaccctg1080ccgcctatctgcaacaacaactgttttcatccaacccagtatccacggccaatgcagctg1140catacctgcaacaacaaceigctacaacagatcctaccagcactcagtcaactagccstgg1200cgaaccctgcctcctacttgcaacagcaacttcttccattcaaccaactggctgtggcga1260acaacgttgcataccagcaacaacatcagctgttgcaagttaacccactggtagtggcta1320acccattggttgccgccttcctacagcaacaacaacaacaactactgccattcaaccaga1380tgtctttgatgaaccctgccttgtcgtggcagcaacccatcgttggacgtgtcttctaga1440ttaaatatgagttgtacttgataaaaaggctttcatgccggcatatgcaacttcctagaa1500ataatcaatatattgattgagatttatcatagtctatttcttttctgtgttgttcatgtt1560aaaaattaaaaacatcatatcataacaaaaggcacatggttggttaattcgtagataata1620ccacattacttcatc犯tccaaatatgtggcaacccaagatcttcaacgttctcatttgg1680gtgtgtgtttgtttataagatacatgtgactgtacatgctcgcagttgagtacttcatag1740gtagtgtgttgtttcaagaaagttattggcacatatgctttgaccatggtcgttcaaaat腦Oaacacccccttttgcaatgcatatatgtgttgataaaagtctatatatgtgttggtgaag1860tctataccaatgcattaattatatgtcgctcgaagtggcgttgcaagggtctattgcaac1920犯ttgtatatacgttggta1939<210>15<211>610<212〉DNA<213>禾本科薏該屬(a/;c/aco;w"-yoW丄.)<400>15atgtctaccaagatatttgttctccttgcgctccttgctctttcagtgagcgctacaact60gcggtcgttattccacaatgctcatttgcttctactactgctacttttccacagttcctc120ccaccattcacagctatggggtttgaacacccagctgtgcaggcctataggttgcaacag180gtgctcgcggtgagcatcttacaacaaccaattgctcaattgcaacaacaaaaacaactc240ctaccagcacttagtcaactagccatggtgaaccctgtcacctacttgcaacaacaagtt300ctaccttccaacccacaggctgtggtgaatgccgctgcatacctacaacaacaacaactg360caacagatcctaccggtgcttagtcaactagctatggcaaacccgatgagttacccggcc420gcctacttgcaactacaacaactgtttccattcaaccaagtagctgtggcgaacgctgct480gcatacctgcaacaacaacaattgtcgcgagtgaatccattggcattggctaacccattg540gttgccgccttcctttgatgaacctgccttgtcatggaagcaacccattgctggtgatgc600tatcttctaa610<210>16<211>547<212>DNA<213>禾本科薏苡屬(CW;c/釘，fl:/oW丄.)<400>16atggctaccaagatatttgtcctccttgtgctccttgctctttcagcgagcgctacaagt60gcggccattattccgcagtgctcacttgctcctagtgctgctattattccgcagttcctc120ccaaccgtttctgctatagggttcgaacacccagctgtgcaggcctataggctacaacag180gcgcttgcgtcaagcatcttgcaacaaccacttatccaattgcaacaacagtcctcggct240catct犯ccatacagaccatcgcagcacaacaacaacaacagttcctgccagcgctcagt300c朋ctagtsgcagcg幼cccagctgcatacttgcaacaacagttgcttgcatccaaccca360tttgctgtggcgaacgctgttgcataccagcaac犯c犯tattcgc犯cagttcctgcca420gcgttcagtcaactggctgtggcacaccctgctgcctatctgcaagcacaactgttttca480tcc犯cccactagctgcggccaataccgctgcatacctgcaacaac犯caactgLcaacag540atcctac547<210>17<211>1942<212>DNA<213>禾本禾斗薏苡屬(Co/x/ac77廳-yoZ",)<400>17misc一feature172…176/note="enciospe:rmaboxmisc一feature201…210/note="opaque_2box"misc一feature335..340/note="CATCbox"TATA—signal402..409CDS501..1442/codonsta;rt=l/product-"alpha-coixin"agcatttgttcatcaattgatgaataacttctcacctaacaaacatgcatggcctaacaa60tgagtataaaactaaaacaaagtcatttgatccaatgagcaagtcaaaacattctgtact120catcatgtttataaagtggaacatttatgatgtggctaaggctaaaacatgtgtaaaggt180gatgtgatcatgcatgtcattcctcatgaatcagaagaattccaatsgaatgcgacactt240ttttcttgtatgtagtggaaactatctttccaacacaaaccatacaatccaatgaaactg300ataact犯atgacataatcgactaggagccatgtcatctctagcttatctgctttttggc360aaaatctaaaatttgaacacacatacatgagcttagacttgtataaatagggctcaaatc420aatagttaatccatcacccattgcactgacagcgattagaaacataccaagtgaaacaca480ctagcagcatcttggcaacaatggctaccaagatatttgtcctcct"tgtgctccttgctc540tttcagcgagcgctacaagtgcggccattattccgcagtgctcacttgctcctagtgctg600ctattattccgcagttcctcccaaccgtttctgcta/tagggttcgaacacccagctgtgc660aggcctataggctacaacaggcgcttgcgtcaagcatcttgcaacaaccacttatccaat720tgc犯c犯cagtcctcggctcatctaaccatacagaccatcgcagcacaacaac犯caac780agttcctgccagcgctcagtcaactagtagcagcgaacccagctgcatatttgcaacaac840ggttgcttgcatccaacccacttgctgtggcgaacgctgttgcataccagcaacaacaat900attcgcaacagttcctgccagcgctcagtcaactggctgtggcaaaccctgctgcctata%0tgcaagcacaactgttttcatccaacccactagctgcagccaataccgctgcatacctgc1020aacaacaacagctacaacagatcctaccagcgctccgtcaactagccctcgcaaaccctg1080ccgcctatttgcaacaacaactgttttcatccaacccactagccgcgaccaatgcagctg1140catacctgcaacaacaacagctacaacagatcctaccagcacttagtcaactagccatgg1200cgaaccctgcctcctacttgcaacaacaacttcttccattcaaccaactggctgtggcga1260acaacgttgcataccagcaacaacatcagctgctgcaagttaacccactggtagtggcta1320acccattggctgccgccttcctacagcagcagcaacaactactactgcaattcaaccaga1380tgtctttgatgaaccctgccttgtcgtgccagcaacccatcgttggacgtgccgtcttct1440agattaaatatgagttgtagttgataaaaaggctttcatgccggcatgtgcaacttccta1500gaaataatcaatatattgattgagatttgtcatagtctatttcttatctgtgttgttcat1560gtgaaaaattaaaaaca/tcatatcataactaaaggcacatggttggttaattcgtagata1620atacaacattacttcatcaatccaaatatgtggcaacccaagatcttcaacgttctcatt1680tgggtgtgtgtttgtttataagatagatgtgactgtacatgctggcagttgagtacttcal了斗Otaggtagtgtgttgtttcaagaaagtgattggcacatatgctttgaccatggccgttcaa1800aataacaccccttttgcaacgaatatatgtgttgataaaagtctatatatgtgttggtca1860agcctataccaatgcattaattatatgtcgctagaagtggcgttgcaagggtctattgca1920acaattgtatatacgttggt1942<210>18<211>1622<212>DNA<213>禾本禾斗薏攻屬(Co/x厶)<400>18misc_feature178..182/n〇te="enciospe:i:ruabox"misc—feature216..225/note="opaque-2box"misc—feature340..345/note="CATCbox"TATA—signal413..420CDS501..1046/codon—start=l/product="alpha-coixingtagcatttgtttattggtcgccgaataatttctcaactaattagtgtgcaagcttcaac60catgagtatgctacaacaaagtcctgaatgcttcgatacacatgagcatgccataacttt120atatactcatcatgtttataaaatagaacatatatgatgtgggcatgtggctaaggttgt180aacatgtgtaaaggtgaagtgatcatgcaggtcattcctcatgtaccaaaagaattccaa240gagaaaatgataatttttcttgcacgtagtggagtctttgcaacaaagagcatataatcc1300aatgaaattgataactaaatgtcaaaatcgactaggtgccatgtcatctctagctcatca360tttcttttggc犯aagccaaaatctgaatagatatacttgagctcttactggtataaata420ggccccaaatcaatagttaaatcaccacccataccattaagagtaatcagaaacttacca480agtgaaacgcgctagcaacaatgtgtaccaagatatttgttcttcttgcgctccttgctc540ttgcagtgagcactacaactgcggtcgttattccgcagtgctcacttgctccacagttcc600tcccaccattcacagctacagggttcgaacacccagttgtgcaggcctataggttacaac660aggtgcttgctgcgaacatcttacaacaaccaattgcccaattacaacaacaacaactcc720tacccacacttagtcaactagccgtcgtgaaccctgccacctacttgcgacaacatcttc780taccttccaacccacaggctgtggtgaatgccggtgcatacctgcaacaacaacagttgc840aacagatcttaccggtgcttagtcaactagctatggcaaacccgaccacctaccccgccg900cctacttacaactgcaacaactgtttccattcaaccaagtagctgtggcgaacgctgctg960catacctgcaacaacaacaattgtcgcgagttaatccattggcattggctaacccattgg1020ttgccgcctttctgcaacaacaataattgcttcaacgagatgtctttgatgaaccctgcc1080ttgtcgtggaagcaacccattgctggtgatgctatcttctaaatcaaatatgagctaaca1140aagctcctataccagcgtgtgtaatttcctagaaataatcaatatattattgagatttat1200cagagtctatttcctattttttcatattaacaactgaaaataccaatttataattaaagg1260aacatgctcgattaatacgtaacaataccacgtttacttcattgatccaaagatgtgcca1320gtctagtctcttgaatgttcttgtttaggtgcgttcctatgtgttcacaagatagataca1380actttacacgcttgcagttgagtacttgcatatgtggtatgttgttttcagaaagtggtg1440ggcacttatgcttgcatggccattcaaatttggtgcacagataccaacttaaatataata1500tgttggggatgaggaccctcggcagcggcaccacacagagcgcaggcaaaaacgaccttc1560gcgtgatccacgcgtccccgctcgaccctttccctcaaaggactgaaccccttggacccg1620U1622<210>19<211>216<212>PRT<213>禾本科薏攻屬(CoZx/acr戸—W)<400〉19MetAlaThrLysliePheAlaLeuLeuValLeuLeuAlaLeuSer51015AlaSerAlaThrThrAlaVallielieProGinCysSerLeuAla202530ProThrValAlalielieSerLysPheLeuProLeuValSerAla354045lieGlyPheGluHisProAlaValArgAlaTyrArgLeuGinGin505560AlaLeuAlaAlaSerlieLeuGinGinProLeuAlaGinLeuGin657075GinGinSerSerAlaHisLeuThrlieGinThrlieAlaAlaGin808590GinGinGinGinPheLeuProAlaLeuSerGinLeuAlaAlaThr95100105AsnProAlaAlaTyrLeuGinGinGinLeuLeuAlaSerAsnPro110115120LeuValValAlaAsnAlaHeAlaTyrGinGinGinGinGinLeu125130135GinGinPheLeuProAlaLeuSerGinLeuAlaValAlaAsnPro140145150AlaAlaTyrLeuGinProGinLeuPheProSerAsnProLeuAla155160165AlaAlaAsnAlaAlaAlaTyrLeuGinGinGinGinLeuGinGin170175180lieLeuProAlaLeuSerGinLeuAlaValAlaAsnProAsnSer185190195TyrLeuGinGinGinGinLeuLeuProPheAsnGinValAlaVal200205210AlaAsnAsnAlaValTyr215<210>20<211>266<212>PRT<213>禾本科薏苡屬(Co/x/acow":/o6/丄.)<400>20MetAlaThrLysliePheAlaLeuLeuValLeuLeuAlaLeuSer51015AlaSerAlaThrThrAlaVallieHeProGinCysSerLeuAla202530ProThrAlaAlalielieGinArgPheLeuProProValSerAla354045lieGlyPheGluHisProAlaValGinAlaTyrArgLeuGinGin505560AlaLeuAlaAlaThrlieLeuGinGinProLeuAlaGinLeuGin657075GinArgSerSerAlaHisLeuThrlieGinThrHeAlaAlaGin8085卯GinGinGinGinGinPheLeuAlaSerLeuSerGinLeuAlaAla95100105AlaAsnProAlaAlaTyrLeuGinGinGinLeuLeuAlaSerAsn110115120ProLeuAlaValAlaAsnAlalieGluTyrGinGinGinGinGin125130135LeuGinGinPheLeuProAlaLeuSerGinLeuValValAlaAsn140145150ProAlaAlaTyrLeuGinSerGinLeuPheProSerAsnProLeu155160165ValValThrAsnAlaAlaAlaTyrLeuGinGinGinGinLeuGin170175180GinHeLeuProAlaLeuSerGinLeuAlaValAlaAsnProAsn185190195SerTyrLeuGinGinGinGinLeuLeuProPheAsnGinValAla200205210ValAlaAsnAsnAlaValTyrGinGinGinTyrGinLeuLeuGin215220225ValAsnProLeuValAlaThrPheLeuGinGinGinGinArgGin230235240LeuLeuProPheAsnGinMetSerLeuMetAsnProAlaLeuSer245250255T卬GinGinProlieValGlyGlyValGlyPhe260265<210>21<211>265<212>PRT<213>禾本科薏苡屬(Co/jc/釘，—6/丄.)<400>21MetAlaThrLysliePheAlaLeuLeuValLeuLeuAlaLeuSer51015GlySerAlaThrThrAlaVallielieProGinCysSerLeuAla202530ProThrAlaAlalielieProArgPheLeuAlaProValSerAla354045HeGlyPheGluHisProAlaValGinAlaTyrArgLeuGinGin505560AlaLeuAlaAlaSerHeLeuGinGinProLeuAlaGinLeuGin657075GinGluSerSerAlaHisLeuAlalieGinThrlieAlaAlaGin808590GinGinGinGinPheLeuProSerLeuSerGinLeuAlaAlaAla95100105AsnProAlaAlaTyrLeuGinGinGinLeuLeuAlaPheAsnPro110115120LeuAlaValAlaAsnAlalieAlaTyrGinGinGinGinGinLeu125130135GinGinPheLeuProAlaLeuSerGinLeuAlaValAlaAsnPro140145150AlaAlaTyrLeuGinSerGinLeuPheProSerAsnProLeuVal155160165ThrAlaAsnValAlaAlaTyrLeuGinGinGinGinLeuGinGin170175180lieLeuProAlaLeuSerGinLeuAlaValAlaAsnProAsnSer185190195TyrLeuGinGinGinGinLeuLeuProPheAsnGinValAlaVal200205210AlaAsnAsnAlaValTyrGinGinGinAsnGinLeuLeuGinVal215220225AsnProLeuAlaAlaAlaPheGinGinGinGinGinArgGinLeu230235240LeuProPheAsnGinMetSerLeuMetAsnProAlaLeuSerTrp245250255GinGinProHeValGlyGlyValGlyPhe260265<210>22<211>265<212>PRT<213>禾本科薏苡屬(Co〖xtor，fl如6".)<400>22MetAlaThrLysliePheAlaLeuLeuValLeuLeuAlaLeuSer51015AlaSerAlaThrThrAlaVallieIkProGinCysSerLeuAla202530ProThrAlaAlalieHeProArgPheLeuProProValSerAla354045ValGlyPheGluHisProAlaValGinAlaTyrArgLeuGinGin505560AlaLeuAlaAlaSerHeLeuGinGinProLeuAspGinLeuGin657075GinGinSerSerAlaHisLeuThrlieGinThrHeAlaAlaGin808590GinGinGinGinPheLeuProSerLeuSerGinLeuAlaAlaAla95100105AsnProAlaAlaTyrLeuGinGinGinLeuLeuAlaPheAsnPro110115120LeuAlaValAlaAsnAlalieAlaTyrHisGinGinGinGinLeu125130135GinGinPheLeuProAlaLeuSerGinLeuAlaValAlaAsnPro140145150AlaThrTyrLeuGinSerGinLeuPheProSerAsnProLeuVal155160165AlaAlaAsnValAlaAlaTyrLeuGinGinGinGinLeuGinGin170175180lieLeuProAlaLeuSerGinLeuAlaValAlaAsnProAsnSer185190195TyrLeuGinGinGinGinLeuLeuProPheAsnGinValAlaVal200205210AlaAsnAsnAlaValTyrGinGinGinHisGinLeuLeuGinVal215220225AsnProLeuAlaAlaAlaPheLeuGinGinGinGinArgGinLeu230235240LeuProPheAsnGinMetSerLeuMetAsnProAlaLeuSerTip245250255GinGinProHeValGlyGlyValGlyPhe260265<210>23<211>265<212>PRT<213>禾本科薏苡屬(Co/xZ釘戸—6".)<400>23MetAlaThrLysliePheAlaLeuLeuValLeuLeuAlaLeuSer51015AlaSerAlaThrThrAlaValHelieProGinCysSerLeuAla202530ProThrAlaAlalielieProArgPheLeuProProValSerAla354045lieGlyPheGluHisProAlaValGinAlaTyrArgLeuGinGin505560AlaLeuAlaAlaSerlieLeuGinGinProLeuAlaGinLeuGin657075GinGinSerSerAlaHisLeuThrlieGinThrHeAlaAlaGin808590GinGinGinGinPheLeuProSerLeuSerGinLeuAlaAlaAla95100105AsnProThrAlaTyrLeuGinGinGinLeuLeuAlaSerAsnPro110115120LeuAlaValAlaAsnAlaHeAlaTyrGinHisGinGinGinLeu125130135GinGinLeuLeuProAlaLeuSerGinLeuAlaValAlaAsnPro140145150AlaAlaTyrLeuGinSerGinLeuPheProSerAsnProLeuVal155160165AlaAlaAsnAlaAlaAlaTyrLeuGinGinGinGinLeuGinGin170175180lieLeuProValLeuSerGinLeuAlaValAlaAspProAsnSer185190195TyrLeuGinGinGinGinLeuLeuProPheAsnGinValAlaVal200205210AlaAsnAsnAlaValTyrGluGinGinHisGinLeuLeuGinVal215220225AsnProLeuAlaAlaAlaPheLeuGinGinGinGinArgGinLeu230235240LeuProPheAsnGinMetSerLeuMetAsnProAlaLeuSerTip245250255GinGinProlieValGlyGlyValGlyPhe260265<210>24<211>264<212>PRT<213>禾本禾斗薏苡屬(Co/;c/"cry附a;/oWL)<400>24MetAlaThrLysliePheAlaLeuLeuValLeuLeuAlaLeuSer51015AlaSerAlaThrThrAlaValHeHeProGinCysSerLeuAla202530ProThrAlaAlalieHeProArgPheLeuProHisValSerAla354045lieGlyPheGluHisProAlaValGinAlaTyrArgLeuGinGin505560AlaLeuAlaAlaSerHeLeuGinGinProLeuAlaGinLeuGin657075GinGinSerSerAlaHisLeuThrlieGinThrlieAlaAlaGin808590GinGinGinPheLeuProSerLeuSerGinGinAlaAlaValAsn95100105ProAlaAlaTyrLeuGinGinGinLeuLeuAlaSerAsnProLeu110115120AlaValProAsnAlalieAlaTyrGinGinGinGinGinLeuGin125130135GinPheLeuProAlaLeuSerGinLeuAlaValAlaAsnProAla140145150AlaTyrLeuGinSerGinLeuPheProCysAsnProLeuValAla155160165AlaAsnAlaAlaAlaTyrLeuGinGinGinGinLeuGinGinlie170175180LeuProAlaLeuSerGinLeuAlaValAlaAsnProAsnSerTyr185190195LeuGinGinGinGinLeuLeuProPheAsnGinValAlaValAla200205210AsnAsnAlaVal丁yrGluGinGinHisGinLeuLeuGinValAsn215220225ProLeuAlaAlaAlaPheLeuGinGinGinGinArgGinLeuLeu230235240ProPheAsnGinMetSerLeuMetAsnProAlaLeuSerTipGin245250255GinProlieValGlyGlyValGlyPhe260<210>25<211>136<212>PRT<213>禾本科薏該屬(Co/x/釘戸一W丄.)<400>25MetAlaThrLysliePheValLeuLeuValLeuLeuAlaLeuSer51015AlaSerAlaThrThrAlaVallieHeProGinCysSerLeuAla202530ProSerAlaAlalielieProGinPheLeuProThrValSerAla354045IkGlyPheGluHisProAlaValGinAlaTyrArgLeuGinGin505560AlaLeuAlaSerSerHeLeuGinGinProLeulieGinLeuGin657075GinGinSerSerAlaHisLeuThrlieGinThrHeAlaThrGin8085卯GinGinGinGinPheLeuProAlaLeuSerGinLeuValAlaAla95100105AsnProAlaAlaTyrLeuGinGinGinLeuPheAlaSerAsnPro110115120LeuSerValAlaAsnAlaValAlaTyrGinGinGinGinTyrSer125130135Gin<210>26<211>312<212>PRT<213>禾本科薏苡屬(Coixr/a^7脂;/oW丄.)<400>26MetAlaThrLysliePheValLeuLeuValLeuLeuAlaLeuSer51015AlaSerAlaThrSerAlaValHelieProGinGlySerLeuAla202530ProThrAlaAlalieHeProGinPheLeuProThrValSerAla354045HeGlyPheGluHisProAlaValGinAlaTyrArgLeuGinGin505560AlaLeuAlaSerSerHeLeuGinGinProLeulieGinLeuGin657075GinGinSerSerAlaHisLeuThrHeGinThrlieAlaAlaGin808590GinGinGinGinPheLeuProAlaLeuSerGinLeuValAlaAla95100105AsnProAlaAlaTyrLeuGluGinGinLeuLeuAlaSerAsnPro110115120LeuAlaValAlaAsnAlaValAlaTyrGinGinGinGinTyrSer125130135GinGinPheLeuProAlaLeuSerGinLeuAlaValAlaAsnPro140145150AlaAlaTyrLeuGinAlaGinLeuPheProSerAsnProLeuAla155160165AlaAlaAsnAlaAlaAlaTyrLeuGinGinArgGinLeuGinGin170175180ValLeuProAlaLeuSerGinLeuValValAlaAsnProAlaAla185190195TyrLeuGinGinGinLeuPheSerSerAsnProValSerThrAla200205210AsnAlaAlaAlaTyrLeuGinGinGinGinLeuGinGinlieLeu215220225ProAlaLeuSerGinLeuAlaMetAlaAsnProAlaSerTyrLeu230235240GinGinGinLeuLeuProPheAsnGinLeuAlaValAlaAsnAsn245250255ValAlaTyrGinGinGinHisGinLeuLeuGinValAsnProLeu260265270ValValAlaAsnProLeuValAlaAlaPheLeuGinGinGinGin275280285GinGinLeuLeuProPheAsnGinMetSerLeuMetAsnProAla290295300LeuSerTrpGinGinProHeValGlyArgValPhe305310<210>27<211>313<212>PRT<213>禾本禾斗薏苡屬(CoZjc/a￡T_yma-_/oWZ.)<400>27MetAlaThrLysliePheValLeuLeuValLeuLeuAlaLeuSer51015AlaSerAlaThrSerAlaAlaHelieProGinCysSerLeuAla202530ProSerAlaAlaHeHeProGinPheLeuProThrValSerAla354045HeGlyPheGluHisProAlaValGinAlaTyrArgLeuGinGin505560AlaLeuAlaSerSerHeLeuGinGinProLeuHeGinLeuGin657075GinGinSerSerAlaHisLeuThrlieGinThrHeAlaAlaGin808590GinGinGinGinPheLeuProAlaLeuSerGinLeuValAlaAla95100105AsnProAlaAlaTyrLeuGinGinArgLeuLeuAlaSerAsnPro110115120LeuAlaValAlaAsnAlaValAlaTyrGinGinGinGinTyrSer125130135GinGinPheLeuProAlaLeuSerGinLeuAlaValAlaAsnPro140145150AlaAlaTyrMetGinAlaGinLeuPheSerSerAsnProLeuAla155160165AlaAlaAsnThrAlaAlaTyrLeuGinGinGinGinLeuGinGin170175180lieLeuProAlaLeuArgGinLeuAlaLeuAlaAsnProAlaAlaTyrLeuGinGinGinLeuPheSerSerAsnProLeuAlaAlaThr200205210AsnAlaAlaAlaTyrLeuGinGinGinGinLeuGinGinlieLeu215220225ProAlaLeuSerGinLeuAlaMetAlaAsnProAlaSerTyrLeu230235240GinGinGinLeuLeuProPheAsnGinLeuAlaValAlaAsnAsn245250255ValAlaTyrGinGinGinHisGinLeuLeuGinValAsnProLeu260265270ValValAlaAsnProLeuAlaAlaAlaPheLeuGinGinGinGin275280285GinLeuLeuLeuGinPheAsnGinMetSerLeuMetAsnProAla290295300LeuSerCysGinGinProHeValGlyArgAlaValPhe305310<210>28<211>181<212>PRT<213>禾本科薏苡屬(CWx/ao^ma^'oWZ.)<400>28MetCysThrLysliePheValLeuLeuAlaLeuLeuAlaLeuAla51015ValSerThrThrThrAlaValVallieProGinCysSerLeuAla202530ProGinPheLeuProProPheThrAlaThrGlyPheGluHisPro354045ValValGinAlaTyrArgLeuGinGinValLeuAlaAlaAsnlie505560LeuGinGinProlieAlaGinLeuGinGinGinGinLeuLeuPro657075ThrLeuSerGinLeuAlaValValAsnProAlaThrTyrLeuArg808590GinHisLeuLeuProSerAsnProGinAlaValValAsnAlaGly95100105AlaTyrLeuGinGinGinGinLeuGinGinlieLeuProValLeu110115120SerGinLeuAlaMetAlaAsnProThrThrTyrProAlaAlaTyr125130135LeuGinLeuGinGinLeuPheProPheAsnGinValAlaValAla140145150AsnAlaAlaAlaTyrLeuGinGinGinGinLeuSerArgValAsn155160165ProLeuAlaLeuAlaAsnProLeuValAlaAlaPheLeuGinGin170175180Gin權利要求1.一種薏苡醇溶蛋白α-coixin基因，其特征在于該基因具有下列核苷酸序列之一(1).具有SEQNO1-18所示的DNA序列；(2).與序列表中序列1-18限定的核酸序列具有95％以上的同源性，且編碼相同功能蛋白質的DNA序列。2.—種根據權利要求l所述的薏苡醇溶蛋白a-coixin基因的編碼蛋白，其特征在于該編碼蛋白具有序列表中序列19-28的氨基酸殘基序列或者是將序列19-28的氨基酸殘基序列經過一個或幾個氨基酸殘基的取代、缺失或添加且具有與序列19-28的氨基酸殘基序列相同活性的由序列19-28衍生的蛋白質。3.根據權利要求1所述的薏苡醇溶蛋白a-coixin基因在在改善植物蛋白質品質中的應用。全文摘要本發明涉及薏苡(Coixlacryma-jobiL.)中醇溶蛋白α-coixin基因，其編碼蛋白以及應用。該基因具有下列核苷酸序列之一(1)具有SEQNO1-18所示的DNA序列；(2)與序列表中序列1-18限定的核酸序列具有95％以上的同源性，且編碼相同功能蛋白質的DNA序列。本發明中發現有些薏苡醇溶蛋白中賴氨酸含量較高，幾乎是玉米醇溶蛋白中賴氨酸含量的2.5倍，在色氨酸含量上也明顯高于玉米醇溶蛋白。我們可以通過提高上述薏苡醇溶蛋白基因的表達量，來改善薏苡種子蛋白的品質，從而達到培育出優質蛋白薏苡的目的。同時也可以利用基因工程的手段，將上述基因導入到玉米胚乳中，來改善玉米種子蛋白的品質，達到培育優質玉米(高賴氨酸玉米)的目的。文檔編號C12N15/82GK101096675SQ20071004119公開日2008年1月2日申請日期2007年5月24日優先權日2007年5月24日發明者孟祥宗,宋任濤,剛王,許政暟,黃彬彬申請人:上海大學