CN101328485B - 编码杜仲的甲羟戊酸途径的酶的基因 - Google Patents
编码杜仲的甲羟戊酸途径的酶的基因 Download PDFInfo
- Publication number
- CN101328485B CN101328485B CN2008101253609A CN200810125360A CN101328485B CN 101328485 B CN101328485 B CN 101328485B CN 2008101253609 A CN2008101253609 A CN 2008101253609A CN 200810125360 A CN200810125360 A CN 200810125360A CN 101328485 B CN101328485 B CN 101328485B
- Authority
- CN
- China
- Prior art keywords
- ala
- ser
- leu
- gly
- val
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Abstract
本发明提供参与杜仲的类异戊二烯化合物生物合成的基因群。本发明还提供参与由杜仲的甲羟戊酸合成异戊烯焦磷酸的基因。本发明的基因编码杜仲的乙酰-CoA C-乙酰转移酶、HMG-CoA合成酶、HMG-CoA还原酶、甲羟戊酸激酶、磷酸甲羟戊酸激酶或甲羟戊酸焦磷酸脱羧酶。
Description
技术领域
本发明涉及参与杜仲的类异戊二烯化合物的生物合成的基因群。
背景技术
作为类异戊二烯化合物之一的聚异戊二烯(橡胶)根据异戊二烯单元的聚合方式的不同而分为顺式和反式。生产长链顺式聚异戊二烯的植物已知有蒲公英、山莴苣等很多植物。巴西橡胶树(Hevea brasiliensis)所生产的顺式聚异戊二烯作为天然橡胶在商业上广泛应用。而关于长链反式聚异戊二烯，已知天然中有胶木等少数植物生产，但商业上并未利用。目前，反式聚异戊二烯是化学合成，应用于高尔夫球外皮、绷带、运动保护器具等中。反式聚异戊二烯是显示低熔点和高弹性的热塑性弹性体，可用作绝缘体。
类异戊二烯化合物通过与碳原子数为5的化合物-异戊烯焦磷酸(IPP)为单元的缩合反应生成。甲羟戊酸途径是类异戊二烯化合物的生物合成中的初期合成途径之一。在动物、植物、菌类中可通过甲羟戊酸途径生物合成IPP。
作为IPP生物合成途径，除了甲羟戊酸途径之外还存在非甲羟戊酸途径。在植物中，在细胞质中是甲羟戊酸途径发挥作用，在色素体中是非甲羟戊酸途径发挥作用。
通过甲羟戊酸途径发挥作用的酶的基因在各种植物中都是公知的。例如Kutki(胡黄连)(Picrorhiza kurrooa)中(根较多含有环烯醚萜苷)登录了乙酰-CoA C-乙酰转移酶(碱基序列在GenBank的登录号为DQ347964，氨基酸序列的登录号为ABC74567)，巴西橡胶树(合成顺式聚异戊二烯)中登录了HMG-CoA合成酶(碱基序列的登录号为AF429389，氨基酸序列登录号为AAS46245)、HMT-CoA还原酶(碱基序列登录号为X54659，氨基酸序列的登录号为P29057)、甲羟戊酸激酶(碱基序列登录号为AF429384，氨基酸序列登录号为AAL18925)、磷酸甲羟戊酸激酶(碱基序列登录号为AF429385，氨基酸序列登录号为AAL18926)、以及甲羟戊酸焦磷酸脱羧酶(碱基序列登录号AF429386，氨基酸序列的登录号为AAL18927)。
中国原产的木本植物杜仲(Eucommia ulmoides Oliver)生产纤维状的长链反式聚异戊二烯。杜仲的叶、树皮和种皮中含有大量反式聚异戊二烯(非专利文献1)。
杜仲的树皮作为滋养强壮和高血压的民间治疗药物，自古以来一直被使用。杜仲中含有类异戊二烯化合物的一种-环烯醚萜的苷(京尼平苷酸)(非专利文献2和3)，其具有降血压作用。
如上所述，杜仲含有如反式聚异戊二烯和环烯醚萜苷等有用的类异戊二烯化合物。
为了制备橡胶高含量的植物，人们希望获得参与杜仲中IPP生物合成途径的基因。以往，在从目标生物种中获得具有某种功能的同源基因时，是利用使用公知基因的DNA片段的杂交筛选、或者是利用公知基因的碱基序列的缩聚PCR与接下来的5’-RACE、3’-RACE和RP-PCR组合的方法。但是，这些方法的准确性低，很麻烦。特别是目标基因的保守区域不明确时，则难于采用这些方法。
对杜仲的基因序列进行了分析(非专利文献4和5)。非专利文献4中报道了参与甲羟戊酸途径的酶之一-HMG-CoA还原酶的基因及其序列。但是参与甲羟戊酸途径的除此之外的酶则未见报道。
非专利文献1：Bamba T，Fukusaki E，Nakazawa Y，and Kobatashi A，In-situ chemical analyses of tran-polyisoprene by histochemical stainingand Fourier transform infrared microapectroscopy in a rubber-producingplant，Eucommia ulmoides Oliver.Planta 215：934-939(2002)
非专利文献2：Kawasaki，T.，Uezono，K.，and Nakazawa，Y.，Antihypertensive mechanism of food for apecified health use：“Eucommialeaf glycoside”and its clinical application，J.Health Sci.，22，29-36(2000)
非专利文献3：Nakmura，T.，Nakazawa，Y.，Onozuka，S.，Tanaka，C.，Yahara，S.and Nohara，T.，Studies on the constituents of Eucommiaulmoides iridoids from the leaves，Natural Medicines，51，275277(1997)
非专利文献4：Jiang J，Kai G，Gao X，and Chen F，Molecular cloningof a HMG-CoA reductase gene from Eucommia ulmoides Oliver.BiosciRep 26：171-181(2006)
非专利文献4：Hou H-W，Zhou Y-T，Mwang K-N，Li W-F，He X-Q，和Cui K-M，ABP1 expression regulated by IAA and ABA is associated withthe cambium periodicity in Eucommia ulmoides Oliv.J Exp Bot57：3857-3867(2006)
发明内容
本发明的目的在于提供参与杜仲的类异戊二烯化合物的生物合成的基因群。
本发明提供参与由杜仲的甲羟戊酸合成异戊烯焦磷酸的基因。上述基因编码选自含有SEQ ID NO.2的氨基酸序列1位-408位氨基酸的蛋白质、含有SEQ ID NO.4的氨基酸序列1位-408位氨基酸的蛋白质、含有SEQID NO.6的氨基酸序列1位-463位氨基酸的蛋白质、含有SEQ ID NO.8的氨基酸序列1位-466位氨基酸的蛋白质、含有SEQ ID NO.12的氨基酸序列1位-629位氨基酸的蛋白质、含有SEQ ID NO.14的氨基酸序列1位-591位氨基酸的蛋白质、含有SEQ ID NO.16的氨基酸序列1位-387位氨基酸的蛋白质、含有SEQ ID NO.18的氨基酸序列1位-506位氨基酸的蛋白质、含有SEQ ID NO.20的氨基酸序列1位-418位氨基酸的蛋白质的蛋白质。
在一个实施方案中，上述基因选自含有SEQ ID NO.1的碱基序列101位-1327位核苷酸的基因、含有SEQ ID NO.3的碱基序列128位-1354位核苷酸的基因、含有SEQ ID NO.5的碱基序列172位-1563位核苷酸的基因、含有SEQ ID NO.7的碱基序列216位-1616位核苷酸的基因、含有SEQ IDNO.11的碱基序列32位-1921位核苷酸的基因、含有SEQ ID NO.13的碱基序列60位-1835位核苷酸的基因、含有SEQ ID NO.15的碱基序列61位-1224位核苷酸的基因、含有SEQ ID NO.17的碱基序列682位-2202位核苷酸的基因、含有SEQ ID NO.19的碱基序列68位-1324位核苷酸的基因。
在另外的实施方案中，上述基因选自含有SEQ ID NO.1的碱基序列1位-1516位核苷酸的基因、含有SEQ ID NO.3的碱基序列1位-1757位核苷酸的基因、含有SEQ ID NO.5的碱基序列1位-1892位核苷酸的基因、含有SEQ ID NO.7的碱基序列1位-1833位核苷酸的基因、含有SEQ ID NO.9的碱基序列1位-1825位核苷酸的基因、含有SEQ ID NO.11的碱基序列1位-2057位核苷酸的基因、含有SEQ ID NO.13的碱基序列1位-2225位核苷酸的基因、含有SEQ ID NO.15的碱基序列1位-1341位核苷酸的基因、含有SEQ ID NO.17的碱基序列1位-2432位核苷酸的基因、含有SEQ IDNO.19的碱基序列1位-1542位核苷酸的基因。
本发明提供参与由杜仲的甲羟戊酸合成异戊烯焦磷酸的基因。
具体实施方式
本发明提供参与由杜仲的甲羟戊酸合成异戊烯焦磷酸(IPP)的基因。IPP的合成通过类异戊二烯化合物的生物合成的初期合成途径-甲羟戊酸途径进行。以下的酶参与甲羟戊酸途径：(1)乙酰-CoA C-乙酰转移酶；(2)3-羟基-3-甲基戊二酰-CoA(HMG-CoA)合成酶；(3)HMG-CoA还原酶；(4)甲羟戊酸激酶；(5)磷酸甲羟戊酸激酶；以及(6)甲羟戊酸焦磷酸脱羧酶(也称为“焦磷酸甲羟戊酸脱羧酶”)。甲羟戊酸途径中，通过(1)乙酰-CoA C-乙酰转移酶，由乙酰-CoA合成乙酰乙酰-CoA，通过(2)HMG-CoA合成酶，由乙酰乙酰-CoA合成HMG-CoA，通过(3)HMG-CoA还原酶，将HMG-CoA转变为甲羟戊酸，通过(4)甲羟戊酸激酶和(5)磷酸甲羟戊酸激酶，将甲羟戊酸转变为甲羟戊酸5-焦磷酸，然后通过(6)甲羟戊酸焦磷酸脱羧酶生成异戊烯焦磷酸(IPP)。
本发明的基因编码选自含有SEQ ID NO.2的氨基酸序列1位-408位氨基酸的蛋白质(以下也称为“SEQ ID NO.2的蛋白质”)；含有SEQ IDNO.4的氨基酸序列1位-408位氨基酸的蛋白质(以下也称为“SEQ IDNO.4的蛋白质”)；含有SEQ ID NO.6的氨基酸序列1位-463位氨基酸的蛋白质(以下也称为“SEQ ID NO.6的蛋白质”)；含有SEQ ID NO.8的氨基酸序列1位-466位氨基酸的蛋白质(以下也称为“SEQ ID NO.8的蛋白质”)；含有SEQ ID NO.12的氨基酸序列1位-629位氨基酸的蛋白质(以下也称为“SEQ ID NO.12的蛋白质”)；含有SEQ ID NO.14的氨基酸序列1位-591位氨基酸的蛋白质(以下也称为“SEQ ID NO.14的蛋白质”)；含有SEQ ID NO.16的氨基酸序列1位-387位氨基酸的蛋白质(以下也称为“SEQ ID NO.16的蛋白质”)；含有SEQ ID NO.18的氨基酸序列1位-506位氨基酸的蛋白质(以下也称为“SEQ ID NO.18的蛋白质”)；含有SEQ IDNO.20的氨基酸序列1位-418位氨基酸的蛋白质(以下也称为“SEQ IDNO.20的蛋白质”)的蛋白质。含有SEQ ID NO.10的1位-466位的氨基酸的蛋白质与SEQ ID NO.8的蛋白质相同。
SEQ ID NO.2的蛋白质和SEQ ID NO.4的蛋白质是(1)乙酰-CoA C-乙酰转移酶。SEQ ID NO.2的蛋白质由含有SEQ ID NO.1的碱基序列101位-1327位的核苷酸的基因编码所得。该基因例如可含有SEQ ID NO.1的全部碱基序列(1位-1516位)。SEQ ID NO.4的蛋白质可由含有SEQ IDNO.3的碱基序列128位-1354位的核苷酸的基因编码获得。该基因例如可含有SEQ ID NO.3的全部碱基序列(1位-1757位)。
SEQ ID NO.6的蛋白质和SEQ ID NO.8的蛋白质是(2)HMG-CoA合成酶。SEQ ID NO.6的蛋白质由含有SEQ ID NO.5的碱基序列172位-1563位的核苷酸的基因编码所得。该基因例如可含有SEQ ID NO.5的全部碱基序列(1位-1892位)。SEQ ID NO.8的蛋白质可由含有SEQ ID NO.7的碱基序列216位-1616位(或SEQ ID NO.9的碱基序列216位-1616位)的核苷酸的基因编码获得。该基因中包含例如含有SEQ ID NO.7的碱基序列1位-1833位的核苷酸的基因、以及含有SEQ ID NO.9的碱基序列1位-1825位的核苷酸的基因。具有SEQ ID NO.9所示碱基序列的基因在其1689位-1825位的区内，相对于具有SEQ ID NO.7所示的碱基序列的基因的1689位-1833位的区，具有6处(SEQ ID NO.7的碱基序列1689位、1709位、1713位、1745位、1756位和1820位)的置换和1处(SEQ ID NO.7的碱基序列1777位)的缺失以及3’末端的聚腺苷部位的缩短。
SEQ ID NO.12的蛋白质和SEQ ID NO.14的蛋白质是(3)HMG-CoA还原酶。SEQ ID NO.12的蛋白质由含有SEQ ID NO.11的碱基序列32位-1921位的核苷酸的基因编码所得。该基因例如可含有SEQ ID NO.11的全部碱基序列(1位-2057位)。SEQ ID NO.14的蛋白质可由含有SEQ IDNO.13的碱基序列60位-1835位的核苷酸的基因编码获得。该基因例如可含有SEQ ID NO.13的全部碱基序列(1位-2225位)。
SEQ ID NO.16的蛋白质是(4)甲羟戊酸激酶。SEQ ID NO.16的蛋白质由含有SEQ ID NO.15的碱基序列61位-1224位的核苷酸的基因编码所得。该基因例如可含有SEQ ID NO.15的全部碱基序列(1位-1341位)。
SEQ ID NO.18的蛋白质是(5)磷酸甲羟戊酸激酶。SEQ ID NO.18的蛋白质由含有SEQ ID NO.17的碱基序列682位-2202位的核苷酸的基因编码所得。该基因例如可含有SEQ ID NO.17的全部碱基序列(1位-2432位)。
SEQ ID NO.20的蛋白质是(6)甲羟戊酸焦磷酸脱羧酶。SEQ IDNO.20的蛋白质由含有SEQ ID NO.19的碱基序列68位-1324位的核苷酸的基因编码所得。该基因例如可含有SEQ ID NO.19的全部碱基序列(1位-1542位)。
本发明的基因只要可以表达与具有上述氨基酸序列的酶功能性同等的酶、或者是可与具有上述碱基序列的基因发挥同样功能即可，可以编码一个或多个氨基酸残基不同的蛋白质，或者可以有一个或多个碱基不同。上述序列的不同可以由于碱基的置换、缺失和/或插入获得，或者发生天然诱变或者是人工诱变(例如定点突变导入法的应用)的任意方式。基因的功能的确定例如可采用如以下实施例2所记载的基本方法的、本领域技术人员通常使用的方法进行。
本发明的基因是本领域技术人员采用通常所使用的方法，按照本说明书记载的序列信息制备探针或引物，以杜仲的染色体DNA或cDNA为模板，通过PCR获得目标片段。当然也可以以RNA为模板，利用反转录PCR。本发明的基因除DNA、RNA等天然多核苷酸之外，也可以是含有人工的核苷酸衍生物的人工分子。本发明的基因还可以是DNA-RNA的嵌合分子。
本发明的基因可以使用本领域技术人员通常所采用的方法，导入到酵母等微生物或植物中。本发明的基因可在宿主中转化，以表达1个或多个、或者是全部。例如其构成可以是：表达存在于甲羟戊酸途径中的一组酶(上述(1)至(6))中的至少一个。
本发明的基因适合用于制备大量含有反式聚异戊二烯或环烯醚萜的基因重组植物(例如杜仲)。本发明的基因例如通过转化杜仲，可用于制备较多含有橡胶的植物。
实施例
以下给出实施例，进一步具体说明本发明，但本发明并不限于此。
(实施例1：杜仲中的EST分析)
(材料)
作为杜仲植物体试样，使用在5月下旬、从爱媛县生名村生长的杜仲标准树采集的当年幼枝的韧皮部(树皮)和木质部。
酵母的突变株文库使用市售的YKO Heterozygous Essential StrainCollection-Glycerol Stocks(Open Biosystems公司)。
(由杜仲中提取RNA)
将约4g杜仲植物体试样(当年枝的韧皮部和木质部)边用液氮冷却，边用乳钵、研磨棒破碎，悬浮于试样的10倍量(w/v)的2×CTAB溶液(2％(w/v)十六烷基三甲基溴化铵(CTAB)、1％(w/v)2-巯基乙醇、0.1M Tris-HCl(pH9.5)、1.4M NaCl、20mM EDTA。将其在65℃下温育10分钟，接着用氯仿/异戊醇处理(洗涤)(反复两次)。向回收的水层中加入1/4(v/v)量的10M LiCl并混合，在-20℃下温育2小时，进行RNA的选择性沉淀。将其离心，将沉淀溶解于适量的Tris-EDTA(TE)缓冲液中，离心分离，回收上清，排除多糖类。将回收的上清进行苯酚处理、苯酚/氯仿处理和氯仿/异戊醇处理，再次通过LiCl进行RNA的选择性沉淀。将沉淀用70％乙醇洗涤，减压干燥，接着溶解于焦碳酸二乙酯(DEPC)处理水中。通过吸光度测定对所得RNA进行定量，通过电泳确认。由约4g木质部中获得0.84mg RNA，由约4g韧皮部中获得2mgRNA(260nm和280nm下的吸收比分别为1.956和1.990)。
(杜仲cDNA文库的制备)
通过日立计测器服务株式会社的G-キッピング方法，由来自杜仲韧皮部和木质部的RNA试样制备cDNA文库。来自韧皮部的cDNA文库的文库大小为3.8×105，插入率为88％(24个样品/琼脂糖凝胶电泳)，全长率86％(对于插入的克隆)。来自木质部的cDNA的文库大小为2.2×105，插入率为79％(24个样品/琼脂糖凝胶电泳)，全长率63％(相对于有插入的克隆)。
(EST序列的序列分析)
在北里大学北里生命科学研究所基因组情报学研究室中，对于来自杜仲韧皮部和木质部的cDNA文库的各约20000个克隆进行序列分析。根据由序列分析得到的序列信息，除去未保有插入的克隆和无法阅读序列的克隆，获得精度高的序列信息。对于韧皮部和木质部的文库，分别得到了16567和16113长度的精度高的EST序列(合计32680)。
对于所得序列进行分组(聚类)(clustering)和注解(annotation)。“分组”是在EST序列中，将同一序列、类似序列分成一组。为了进行分组，使用NTT软件的VISUALBIO clustering。“注解”是通过与已知基因比较，对EST序列进行注解。注解时使用采用NCBI BLAST的同源性检索。检索时所使用的数据库是nr(All non-redundant GenBank CDStranslations+PDB+SwissProt+PIR(肽序列数据库))。
根据分组和注解所得的信息，发现了推定为编码乙酰-CoA C-乙酰转移酶、HMG-CoA合成酶、HMG-CoA还原酶、甲羟戊酸激酶和甲羟戊酸焦磷酸脱羧酶的EST序列。这些酶均是参与杜仲的类异戊二烯化合物的生物合成中初期合成途径的酶。
(实施例2：编码乙酰-CoA C-乙酰转移酶的基因1)
(全长cDNA的获得)
通过3’-RACE(cDNA末端的快速扩增，RACE)，由通过实施例1的分析得到的序列确定3’末端一侧的序列，获得全长cDNA。
3’-RACE使用3’-Full RACE Core Set(タカラバイオ株式会社制备)。反转录反应中使用oligo-dT引物。PCR扩增使用与oligo-dT的引物和与已知序列的一部分为相同的序列的有义引物。有义引物根据推定为实施例1所得的编码乙酰-CoA C-乙酰转移酶的EST序列信息设计。以实施例1所得的RNA为模板，第1次是使用N192-82-tree_1968_3R_S1(SEQ IDNO.21)作为有义引物，第2次是使用N192-82-tree_1968_3R_s2(SEQ IDNO.22)作为有义引物，按照说明书进行PCR。将反转录反应和由PCR得到的扩增片段TA克隆到pT7Blue载体中，进行序列分析。
所得全长cDNA具有SEQ ID NO.1的碱基序列。SEQ ID NO.1所述的碱基序列146位-1299位的核苷酸所示的区域的碱基序列与Kutki的乙酰-CoA C-乙酰转移酶(GenBank登录号DQ347964)具有82％的同源性。开放阅读框是101位-1327位。由该cDNA编码的推定的氨基酸序列如SEQID NO.2所示。SEQ ID NO.2的全长氨基酸序列(1位-408位)与Kutki的乙酰-CoA C-乙酰转移酶(登录号ABC74567)具有89％的同源性。
(使用酵母突变株文库的互补性试验)
使用用酵母突变株文库进行的互补性试验确认所得cDNA的功能。该互补性试验如下进行。
使用附加有与pYES2载体(invitrogen制备)的多克隆位点的40bp为相同序列的有义引物和反义引物，进行PCR(95℃下将5分钟作为1个循环；然后95℃下将1分钟、54℃下将1分钟和72℃下将1分钟作为30个循环；在72℃下将7分钟作为1个循环；以及4℃下将∞作为1个循环)，扩增目标基因的翻译区序列。
将该扩增片段和用限制酶处理成直链状的pYES2载体导入到Ura-的酵母株Y22800(EUROSCARF制备)中。作为分析对象的乙酰-CoA C-乙酰转移酶基因对于酵母的生长来说是必须的，因此，作为互补试验的背景，该酵母使用只有一侧的对位基因缺失目标基因的杂合二倍体。基因导入时使用Frozen-EZ Yeast Transformation II(ZYMO RESEARCH制备)。将转化后的酵母接种于极限完全培养基(无尿嘧啶)，在30℃下培养。将生长的集落接种于新的极限完全培养基(无尿嘧啶)中，接着在30℃下培养。酵母突变株是尿嘧啶需求性，pYES2载体中含有URA3基因(编码乳清苷5’-磷酸脱羧酶)的序列，因此只有转化的菌体可以生长。生长的菌体通过PCR检查其是否含有插入片段，接着，将该菌体转移至YPDA培养基中，在30℃下培养。将其转移至孢子形成培养基，在25℃下培养，将所得孢子进行四分体分析。
插入到pYES2载体的多克隆位点上的目标基因上的上游存在GAL1启动子，因此，目标基因只在半乳糖存在下诱导表达。因此，判定是否在YPD培养基或YPG培养基(碳源：半乳糖)上进行生长。在YPD培养基中，来自突变株的二分体致死，只有来自野生型的二分体生长。而在YPG培养基(碳源：半乳糖)上，具有GAL1启动子的pYES2载体诱导插入基因的表达，因此，如果功能互补，则来自突变株的可以存活，三分体或四分体生长。这样，通过进行互补的确认，可以进行目标基因的功能确定。
并且，将由四分体分析得到的来自八孢子囊的集落(可认为是单倍体)影印到(A)YPG培养基(碳源：半乳糖)、(B)极限完全培养基(碳源：半乳糖、不含有尿嘧啶)、(C)YPG培养基(碳源：半乳糖，含有抗生素G418)、或(D)YPG培养基(碳源：葡萄糖)上。
保有质粒的培养株在(B)的极限完全培养基(碳源：半乳糖，不含尿嘧啶)中生长。突变株具有G418抗性基因，因此可认为在(C)YPG培养基(碳源：半乳糖，含有抗生素G418)中生长。由这些培养基中的生长结果可以判别出保有质粒的单倍体突变株。在(D)的YPD培养基(碳源：葡萄糖)中，保有质粒的突变株(单倍体)应无法生长，但实际上难以判定其生长的有无。因此通过筛选判别生长的有无。
本实施例中，使用N219-36-tree-1968_S(SEQ ID NO.23)作为有义引物，使用N219-36-tree_1968_a(SEQ ID NO.24)作为反义引物，以上述“完全cDNA的获得”中得到的cDNA作为模板，进行PCR，扩增目标翻译区的序列。将该扩增片段供给上述互补性试验。在四分体分析中观察到在YPG培养基(碳源：半乳糖)上的生长。并且通过使用来自八孢子囊的集落的生长观察，观察到在(B)极限完全培养基(碳源：半乳糖，不含尿嘧啶)上的生长。结果，通过互补，确定了乙酰-CoA C-乙酰转移酶基因的功能。
(实施例3：编码乙酰-CoA C-乙酰转移酶的基因2)
使用根据实施例1得到的推定为编码乙酰-CoA C-乙酰转移酶的EST序列的信息设计的N192-84-tree_11012_3R_s1(SEQ ID NO.25)(第1次)和N192-84-tree_11012_3R_s2(SEQ ID NO.26)(第2次)作为3’-RACE用的有义引物，除此之外与实施例2同样，得到全长cDNA。
所得全长cDNA具有SEQ ID NO.3的碱基序列。SEQ ID NO.3的碱基序列173位-1305位的核苷酸所示的区域的碱基序列与Kutki的乙酰-CoAC-乙酰转移酶(登录号DQ347964)具有83％的同源性。开放阅读框为128位-1354位。由该cDNA编码的推定氨基酸序列如SEQ ID NO.4所示。SEQID NO.4的全长氨基酸序列(1位-408位)与Kutki的乙酰-CoA C-乙酰转移酶(登录号ABC74567)具有87％的同源性。
本实施例中，使用N219-36-tree_11012_s(SEQ ID NO.27)作为有义引物，使用N-219-36-tree_11012_a(SEQ ID NO.28)作为反义引物，以上述所得cDNA为模板，除此之外与实施例2同样进行PCR，扩增目标翻译区的序列。将该扩增片段与实施例2同样供给互补性试验，得到了与实施例2同样的结果。在四分体分析中观察到在YPG培养基(碳源：半乳糖)上的生长。并且，通过使用来自八孢子囊的集落的生长观察，观察到在(B)极限完全培养基(碳源：半乳糖，不含尿嘧啶)上的生长。结果，通过互补确定了乙酰-CoA C-乙酰转移酶基因的功能。
(实施例4：编码HMG-CoA合成酶的基因1)。
使用基于实施例1所得的推定为编码HMG-CoA合成酶的EST序列的信息设计的N192-71-tree_6098-3R_s1(SEQ ID NO.29)(第1次)和N192-71-tree_6098_3R_s2(SEQ ID NO.30)(第2次)作为3’-RACE的有义引物，除此之外与实施例2同样，得到全长cDNA。
所得的全长cDNA具有SEQ ID NO.5的碱基序列。SEQ ID NO.5的碱基序列172位-1187位的核苷酸所示的区域和1390位-1439位的核苷酸所示的区域的碱基序列，分别与巴西橡胶树的HMG-CoA合成酶(登录号AF429389)具有83％和86％的同源性。开放阅读框是172位-1563位。由该cDNA编码的推定氨基酸序列如SEQ ID NO.6所示。SEQ ID NO.6的全长氨基酸序列(1位-463位)与巴西橡胶树的HMG-CoA合成酶(登录号AAS46245)具有85％的同源性。
本实施例中，有义引物使用N219-36-tree_6098_s(SEQ ID NO.31)，反义引物使用N219-36-tree_6098_a(SEQ ID NO.32)，以上述所得cDNA为模板，除此之外与实施例2同样进行PCR，扩增目标翻译区的序列。与实施例2同样，将该扩增片段供给互补性试验。突变株使用Ura-的HMG-CoA合成酶缺损株Y26527(EUROSCARF制备)。在四分体分析中观察到在YPG培养基(碳源：半乳糖)上的生长。并且，在使用来自八孢子囊的集落的生长观察中，观察到在(B)极限完全培养基(碳源：半乳糖，不含尿嘧啶)上的生长。结果，通过互补确定了HMG-CoA合成酶基因的功能。
(实施例5：编码HMG-CoA合成酶的基因2)
使用实施例1所得的推定编码HMG-CoA合成酶的EST序列的信息设计的N192-36-tree_10370_3R_s1(SEQ ID NO.33)(第1次)和N192-36-tree_10370_3R_s2(SEQ ID NO.34)(第2次)，作为3’-RACE用的有义引物，除此之外与实施例2同样得到全长cDNA。得到了两个全长cDNA。
所得的第一全长cDNA具有SEQ ID NO.7的碱基序列。SEQ ID NO.7的碱基序列234位-1496位的核苷酸所示区域的碱基序列与巴西橡胶树的HMG-CoA合成酶(登录号AF429389)具有81％的同源性。所得第二全长cDNA具有SEQ ID NO.9的碱基序列。该第二cDNA在其1689位-1825位的区域内，对应第一cDNA的1689位-1833位的区域有6处(第一cDNA的1689位、1709位、1713位、1745位、1756位和1820位)置换和1处(第一cDNA的1777位)缺失、以及3’末端的聚腺苷部位的缩短。SEQ ID NO.9的碱基序列也与SEQ ID NO.7的碱基序列同样，与巴西橡胶树的HMG-CoA合成酶(登录号AF429389)具有81％的同源性。开放阅读框在两个碱基序列均为216位-1616位。由这些cDNA编码的推定氨基酸序列分别如SEQ ID NO.8和10所示，它们是相同的序列。该推定氨基酸序列在SEQ ID NO.8的6位-452位的氨基酸区域中，与巴西橡胶树的HMG-CoA合成酶(登录号AAS46245)具有84％的同源性。
本实施例中，使用N219-36-tree_10370_s(SEQ ID NO.35)作为有义引物，使用N219-36-tree_10370_a(SEQ ID NO.36)作为反义引物，以上述所得cDNA作为模板，除此之外与实施例2同样进行PCR，扩增目标翻译区的序列。与实施例2同样，将该扩增片段供给互补性试验。突变株使用Ura-的HMG-CoA合成酶缺损株Y26527(EUROSCARF制备)。在四分体分析中观察到YPG培养基(碳源：半乳糖)上的生长。并且，在使用来自八孢子囊的集落的生长观察中，观察到在(B)极限完全培养基(碳源：半乳糖，不含尿嘧啶)上的生长。结果，通过互补，确定了HMG-CoA合成酶基因的功能。
(实施例6：编码HMG-CoA还原酶的基因1)
关于编码HMG-CoA还原酶的基因，为了获得全长cDNA，采用5’RACE、3’RACE和反转录反应组合的方法。该方法中，使用5’-FullRACE Core Set(TaKaRa制备)和3’-Full RACE Core Set(TaKaRa制备)。根据实施例1所得的推定为编码HMG-CoA还原酶的EST序列设计N192-45-tree_8220_5R_a1(SEQ ID NO.37)(第一次)和N192-45-tree_8220_5R_a2(SEQ ID NO.38)(第二次)作为5’RACE用的反义引物，N192-45-tree_8220_5R_3R_s1(SEQ ID NO.39)(第一次)和N192-45-tree_8220_5R_3R_s2(SEQ ID NO.40)(第二次)作为3’-RACE用的有义引物，N192-45-tree_8220_5R_p(SEQ ID NO.41)作为反转录反应用。以实施例1所得的RNA为模板，与实施例2同样地，将它们反应所得的扩增片段TA克隆到pTCBlue载体上，进行序列分析。
所得全长cDNA具有SEQ ID NO.11的碱基序列。SEQ ID NO.11的碱基序列260位-303位的核苷酸所示区域、773位-936位的核苷酸所示区域、以及1148位-1845位的核苷酸所示的的区域的碱基序列分别与苹果(Malus x domestica)的HMG-CoA还原酶(登录号AF315713)具有88％、82％和81％的同源性。开放阅读框是32位-1921位。由该cDNA编码的推定氨基酸序列如SEQ ID NO.12所示。在SEQ ID NO.12的33位-612位的氨基酸区域中，该推定氨基酸序列与喜树(Camptotheca acuminata)的HMG-CoA还原酶(登录号P48021)具有78％的同源性。SEQ ID NO.12的33位-612位的氨基酸区域与非专利文献4的HMG-CoA还原酶(登录号AAV54051)具有73％的同源性。因此，可以认为序列号11的碱基序列是来自与非专利文献4的基因不同的基因座的HMG-CoA还原酶基因。
(实施例7：编码HMG-CoA还原酶的基因2)
根据实施例1所得的推定为编码HMG-CoA还原酶的EST序列设计N192-47-tree_13453_5R_a1(SEQ ID NO.42)(第一次)和N192-47-tree_13453_5R_a2(SEQ ID NO.43)(第二次)作为5’RACE用的反义引物，N192-47-tree_13453_5R_3R_s1(SEQ ID NO.44)(第一次)和N192-47-tree_13453_5R_3R_s2(SEQ ID NO.45)(第二次)作为3’-RACE用的有义引物，N192-47-tree_13453_5R_p(SEQ ID NO.46)作为反转录反应用，除此之外与实施例6同样，得到全长cDNA。
所得全长cDNA具有SEQ ID NO.13的碱基序列。SEQ ID NO.13的碱基序列169位-373位的核苷酸所示区域、751位-1600位的核苷酸所示区域、以及1637位-1720位的核苷酸所示的的区域的碱基序列，分别与喜树的HMG-CoA还原酶(登录号U72145)具有79％、80％和89％的同源性。开放阅读框是60位-1835位。由该cDNA编码的推定氨基酸序列如SEQ IDNO.14所示。在SEQ ID NO.14的1位-588位的氨基酸区域中，该推定氨基酸序列与烟草(Nicotiana tabacum)的HMG-CoA还原酶(登录号AAB87727)具有78％的同源性。SEQ ID NO.14的1位-589位的氨基酸区域与非专利文献4的HMG-CoA还原酶(登录号AAV54051)具有75％的同源性。因此，可以认为序列号13的碱基序列也是来自与非专利文献4的基因不同的基因座的HMG-CoA还原酶基因。
(实施例8：编码甲羟戊酸激酶的基因)
3’-RACE用的有义引物使用基于实施例1所得的推定为编码甲羟戊酸激酶的EST序列的信息设计的tree 1729引物(SEQ ID NO.47)(第一次)和tree 1729引物套(SEQ ID NO.48)(第二次)，除此之外与实施例2同样，得到全长cDNA。
所得全长cDNA具有SEQ ID NO.15的碱基序列。SEQ ID NO.15的碱基序列616位-1104位的核苷酸所示区域的碱基序列与巴西橡胶树的甲羟戊酸激酶(登录号AF429384)具有83％的同源性。开放阅读框是61位-1224位。由该cDNA编码的推定氨基酸序列如SEQ ID NO.16所示。SEQ IDNO.16全长氨基酸序列(1位-387位)与巴西橡胶树的甲羟戊酸激酶(登录号AAL18925)具有77％的同源性。
本实施例中，使用N219-36-tree_1729_s(SEQ ID NO.49)作为有义引物，使用N219-36-tree_1729_a(SEQ ID NO.50)作为反义引物，以上述所得cDNA作为模板，除此之外与实施例2同样进行PCR，扩增目标翻译区的序列。将该扩增片段与实施例2同样地供给互补性试验。突变株使用Ura-的甲羟戊酸激酶缺损株Y20794(EUROSCARF制备)。在四分体分析中观察到在YPG培养基(碳源：半乳糖)上的生长。并且，在使用来自八孢子囊的集落的生长观察中，观察到在(B)极限完全培养基(碳源：半乳糖，不含尿嘧啶)上的生长。结果，通过互补，确定了甲羟戊酸激酶基因的功能。
(实施例9：编码磷酸甲羟戊酸激酶的基因)
以实施例1所得cDNA文库作为模板，使用以下引物进行PCR，获得部分序列。该简并PCR中使用的引物如下：第一次使用S_No.243_P.10_1(SEQ ID NO.51)作为有义引物和使用AS_No.243_P.10_1(SEQ IDNO.52)作为反义引物；第一次使用S_No.243_P.10_2(SEQ ID NO.53)作为有义引物和使用AS_No.243_P.10_2(SEQ ID NO.54)作为反义引物，引物的设计基于BAD93946(拟南芥，Arabidopsis thaliana)、AAL18926(巴西橡胶树，Hevea brasiliensis)、CAB52264(粟酒裂殖酵母，Schizosaccharomyces pombe)、NP593421(粟酒裂殖酵母，Schizosaccharomyces pombe)、P24521(酿酒酵母，Saccharomycescerevisiae)和XP329795(粗糙脉孢霉，Neurospora crassa)的六个氨基酸序列的保守区域进行。PCR的条件如下：
(第一次PCR)
95℃5分钟进行一个循环；将95℃1分钟、55℃1分钟、72℃1分30秒进行30个循环；72℃7分钟进行一个循环；4℃∞进行一个循环；
(第二次PCR)
95℃5分钟进行一个循环；将95℃1分钟、55℃1分钟、72℃1分30秒进行30个循环；72℃7分钟进行一个循环；4℃∞进行一个循环；
上述部分序列中未得到3’末端和5’末端的碱基序列信息，因此，以实施例1所得的RNA作为模板，进行5’-RACE、3’-RACE和反转录反应。该方法中使用5’-Full RACE Core Set(Takara)和3’-Full RACE Core Set(Takara)。根据所得部分片段的信息设计第一次的No.243_P.51_race_a1(SEQ ID NO.56)和第二次的No.243_P.51_race_a2(SEQ ID NO.55)作为5’-RACE用的反义引物，第一次的No.243_P.51_race_s1(SEQ IDNO.57)和第二次的No.243_P.51_race_s2(SEQ ID NO.58)作为3’-RACE用的有义引物，No.243_P.51_race_p(SEQ ID NO.59)作为反转录反应用。由此得到3’末端和5’末端的碱基序列信息。
根据3’末端和5’末端的碱基序列信息，设计cDNA文库筛选用探针，使用SEQ ID NO.60的引物作为有义引物，使用SEQ ID NO.61的引物作为反义引物，进行PCR。以所得片段作为探针，筛选实施例1所得的cDNA文库。将所得片段TA克隆到pT7Blue载体上，进行序列分析。
所得cDNA片段的全部碱基序列如SEQ ID NO.17所示。SEQ IDNO.17的碱基序列681位-886位的核苷酸所示的区域、922位-1000位的核苷酸所示的区域、1042位-1285位的核苷酸所示的区域、1339位-1732位的核苷酸所示的区域、以及1807位-2054位的核苷酸所示区域的碱基序列分别与巴西橡胶树的磷酸甲羟戊酸激酶(登录号AF429385)具有83％、84％、81％、81％和78％的同源性。开放阅读框为682位-2202位。由该cDNA编码的推定氨基酸序列如SEQ ID NO.18所示。SEQ ID NO.18的全长氨基酸序列(1位-506位)与巴西橡胶树的磷酸甲羟戊酸激酶(登录号AAL18926)具有78％的同源性。
本实施例中，使用SEQ ID NO.62的引物作为有义引物，使用SEQ IDNO.63的引物作为反义引物，以上述所得cDNA作为模板，除此之外与实施例2同样进行PCR，扩增目标翻译区的序列。将该扩增片段与实施例2同样供给互补性试验。突变株使用Ura-的磷酸甲羟戊酸激酶缺损株Y20806(EUROSCARF制备)。在四分体分析中观察到在YPG培养基(碳源：半乳糖)上的生长。并且，在使用来自八孢子囊的集落的生长观察中，观察到在(B)极限完全培养基(碳源：半乳糖，不含尿嘧啶)上的生长。结果，通过互补，确定了磷酸甲羟戊酸激酶基因的功能。
(实施例10：编码甲羟戊酸焦磷酸脱羧酶的基因)
使用基于实施例1所得的推定为甲羟戊酸焦磷酸脱羧酶的EST序列的信息设计的N192-102-tree_13103_3R_s1(SEQ ID NO.64)(第一次)和N192-102-tree_13103_3R_s2(SEQ ID NO.65)(第二次)作为3’-RACE用的有义引物，除此之外与实施例2同样，得到全长cDNA。
所得全长cDNA具有SEQ ID NO.19的碱基序列。SEQ ID NO.1的碱基序列100位-1325位的核苷酸所示区域的碱基序列与巴西橡胶树的甲羟戊酸焦磷酸脱羧酶(登录号AF429386)具有80％的同源性。开放阅读框为68位-1324位。由该cDNA编码的推定氨基酸序列如SEQ ID NO.20所示。在SEQ ID NO.20的5位-418位的氨基酸区域中，该推定氨基酸序列与巴西橡胶树的甲羟戊酸焦磷酸脱羧酶(登录号AAL18927)具有84％的同源性。
本实施例中，使用N219-36-tree_13103_s(SEQ ID NO.66)作为有义引物，使用N219-36-tree_13103_a(SEQ ID NO.67)作为反义引物，以上述所得cDNA作为模板，除此之外与实施例2同样进行PCR，扩增目标翻译区的序列。将该扩增片段与实施例2同样供给互补性试验。突变株使用Ura-的甲羟戊酸焦磷酸脱羧酶缺损株Y25418(EUROSCARF制备)。在四分体分析中观察到在YPG培养基(碳源：半乳糖)上的生长。并且，在使用来自八孢子囊的集落的生长观察中，观察到在(B)极限完全培养基(碳源：半乳糖，不含尿嘧啶)上的生长。结果，通过互补，确定了甲羟戊酸焦磷酸脱羧酶基因的功能。
产业实用性
本发明提供编码杜仲的类异戊二烯化合物的生物合成中的初期合成途径-甲羟戊酸途径的各种酶的基因。本发明的基因适合用于制备大量含有反式异戊二烯或环烯醚萜的基因重组植物(例如杜仲)。
序列表
<110>日立造船株式会社
大阪大学
<120>编码杜仲的甲羟戊酸途径的酶的基因
<130>P2-08H03050
<160>67
<170>PatentIn version 3.1
<210>1
<211>1516
<212>DNA
<213>杜仲
<220>
<221>CDS
<222>(101)..(1327)
<223>乙酰-CoA C乙酰转移酶
<400>1
cgattatcga ctctataggg aatccttaaa tttgaaactt ctcaccaacc tcttgtgatc 60
tcggagcgct cccaatcaac cggaggtttt aggtgctctc atg gct cca gca gca 115
Met Ala Pro Ala Ala
1 5
gca aca gag tgc tca gat tcc ata aag ccc aga gat gtt tgt att gtt 163
Ala Thr Glu Cys Ser Asp Ser Ile Lys Pro Arg Asp Val Cys Ile Val
10 15 20
gga gtt gca cga aca cca atg ggt gct ttt ctt gga tct ctt tca tca 211
Gly Val Ala Arg Thr Pro Met Gly Ala Phe Leu Gly Ser Leu Ser Ser
25 30 35
tta cca gcc acc aag ctt gga tct ata gct att gaa agt gct cta aaa 259
Leu Pro Ala Thr Lys Leu Gly Ser Ile Ala Ile Glu Ser Ala Leu Lys
40 45 50
aga gca aat gtt gac cca tca ctt gta gaa gaa gtg ttc ttt ggt aat 307
Arg Ala Asn Val Asp Pro Ser Leu Val Glu Glu Val Phe Phe Gly Asn
55 60 65
gtt ctt agt gca aat ttg ggt cag gct cca gct aga caa gca gca ctt 355
Val Leu Ser Ala Asn Leu Gly Gln Ala Pro Ala Arg Gln Ala Ala Leu
70 75 80 85
ggt gct gga ata cct agc aca gtg gtt tgc acc acc ata aac aaa gtt 403
Gly Ala Gly Ile Pro Ser Thr Val Val Cys Thr Thr Ile Asn Lys Val
90 95 100
tgt gct tca gga atg aaa gca act atg ctt gct gct cag agc atc cag 451
Cys Ala Ser Gly Met Lys Ala Thr Met Leu Ala Ala Gln Ser Ile Gln
105 110 115
ttg ggc atc agt gat gtt gtc ata gct ggt ggg atg gaa agc atg tct 499
Leu Gly Ile Ser Asp Val Val Ile Ala Gly Gly Met Glu Ser Met Ser
120 125 130
aat gta ccc aag tac ctt gca gaa gca agg aag gga tcc cga ctt gga 547
Asn Val Pro Lys Tyr Leu Ala Glu Ala Arg Lys Gly Ser Arg Leu Gly
135 140 145
cat gat act ctt gtt gat gga atg ttg aaa gat ggt ttg tgg gat gta 595
His Asp Thr Leu Val Asp Gly Met Leu Lys Asp Gly Leu Trp Asp Val
150 155 160 165
tat gat gac ttt gga atg gga gtg tgt gct gaa cta tgt gct gag cat 643
Tyr Asp Asp Phe Gly Met Gly Val Cys Ala Glu Leu Cys Ala Glu His
170 175 180
cat gga gta aca aga gat gaa cag gac aac tat gcc atc caa agc ttt 691
His Gly Val Thr Arg Asp Glu Gln Asp Asn Tyr Ala Ile Gln Ser Phe
185 190 195
gag cgt ggt att gct gct aaa gat tgt gat gca ttt gtt tgg gaa ata 739
Glu Arg Gly Ile Ala Ala Lys Asp Cys Asp Ala Phe Val Trp Glu Ile
200 205 210
gtt cca gtt gaa gtt tcc ggt gga aga gga aga cca tct aca att gtt 787
Val Pro Val Glu Val Ser Gly Gly Arg Gly Arg Pro Ser Thr Ile Val
215 220 225
gac aag gat gaa ggt cta gga aag ttt gat gct tca aag ttg aga aag 835
Asp Lys Asp Glu Gly Leu Gly Lys Phe Asp Ala Ser Lys Leu Arg Lys
230 235 240 245
ctc cga cca agt ttc aaa gaa act ggt ggt tct gtt act gct ggc aat 883
Leu Arg Pro Ser Phe Lys Glu Thr Gly Gly Ser Val Thr Ala Gly Asn
250 255 260
gct tct agt atc agt gat ggt gct gca gca ctc gtt tta gtt tgt gga 931
Ala Ser Ser Ile Ser Asp Gly Ala Ala Ala Leu Val Leu Val Cys Gly
265 270 275
gag aag gca ctt aag ctt ggt ctg caa gtg att gca aaa atc aca gga 979
Glu Lys Ala Leu Lys Leu Gly Leu Gln Val Ile Ala Lys Ile Thr Gly
280 285 290
tat gct gac gct gct cag gca cct gag ttg ttt aca act tct cca gct 1027
Tyr Ala Asp Ala Ala Gln Ala Pro Glu Leu Phe Thr Thr Ser Pro Ala
295 300 305
ctt gcc ata ccg aaa gct atc tca aat gct ggc ata gag gct tct caa 1075
Leu Ala Ile Pro Lys Ala Ile Ser Asn Ala Gly Ile Glu Ala Ser Gln
310 315 320 325
gtt gat tac tac gaa att aat gaa gcc ttt gcg gtt gtt gct ctg gca 1123
Val Asp Tyr Tyr Glu Ile Asn Glu Ala Phe Ala Val Val Ala Leu Ala
330 335 340
aat cag aaa ctt ctt gga tta gct ccg gaa aaa gta aat gta cat ggt 1171
Asn Gln Lys Leu Leu Gly Leu Ala Pro Glu Lys Val Asn Val His Gly
345 350 355
gga gct gta tct ctg ggg cat cct cta ggc tgc agc gga gca cgt att 1219
Gly Ala Val Ser Leu Gly His Pro Leu Gly Cys Ser Gly Ala Arg Ile
360 365 370
gta gtc aca ctt cta ggg gtg ttg agg cag aag aat ggg aaa tat gga 1267
Val Val Thr Leu Leu Gly Val Leu Arg Gln Lys Asn Gly Lys Tyr Gly
375 380 385
gta agt ggt gtc tgc aat ggg gga ggc ggt gca tct gcc ctt gtc gta 1315
Val Ser Gly Val Cys Asn Gly Gly Gly Gly Ala Ser Ala Leu Val Val
390 395 400 405
gag ctt gtc taa tccctaccta ttgaggctaa ttaagacttt aagaagaaat 1367
Glu Leu Val
tagcttcata tcaacttcag ttcagttcag aagatgtgca atttttttct ctccagtttg 1427
ttctcttcac taataaatgt attcaggttg taaacgagtt aatatatatt caataaatat 1487
ttccaatgtt caccgaaaaa aaaaaaaaa 1516
<210>2
<211>408
<212>PRT
<213>杜仲
<400>2
Met Ala Pro Ala Ala Ala Thr Glu Cys Ser Asp Ser Ile Lys Pro Arg
1 5 10 15
Asp Val Cys Ile Val Gly Val Ala Arg Thr Pro Met Gly Ala Phe Leu
20 25 30
Gly Ser Leu Ser Ser Leu Pro Ala Thr Lys Leu Gly Ser Ile Ala Ile
35 40 45
Glu Ser Ala Leu Lys Arg Ala Asn Val Asp Pro Ser Leu Val Glu Glu
50 55 60
Val Phe Phe Gly Asn Val Leu Ser Ala Asn Leu Gly Gln Ala Pro Ala
65 70 75 80
Arg Gln Ala Ala Leu Gly Ala Gly Ile Pro Ser Thr Val Val Cys Thr
85 90 95
Thr Ile Asn Lys Val Cys Ala Ser Gly Met Lys Ala Thr Met Leu Ala
100 105 110
Ala Gln Ser Ile Gln Leu Gly Ile Ser Asp Val Val Ile Ala Gly Gly
115 120 125
Met Glu Ser Met Ser Asn Val Pro Lys Tyr Leu Ala Glu Ala Arg Lys
130 135 140
Gly Ser Arg Leu Gly His Asp Thr Leu Val Asp Gly Met Leu Lys Asp
145 150 155 160
Gly Leu Trp Asp Val Tyr Asp Asp Phe Gly Met Gly Val Cys Ala Glu
165 170 175
Leu Cys Ala Glu His His Gly Val Thr Arg Asp Glu Gln Asp Asn Tyr
180 185 190
Ala Ile Gln Ser Phe Glu Arg Gly Ile Ala Ala Lys Asp Cys Asp Ala
195 200 205
Phe Val Trp Glu Ile Val Pro Val Glu Val Ser Gly Gly Arg Gly Arg
210 215 220
Pro Ser Thr Ile Val Asp Lys Asp Glu Gly Leu Gly Lys Phe Asp Ala
225 230 235 240
Ser Lys Leu Arg Lys Leu Arg Pro Ser Phe Lys Glu Thr Gly Gly Ser
245 250 255
Val Thr Ala Gly Asn Ala Ser Ser Ile Ser Asp Gly Ala Ala Ala Leu
260 265 270
Val Leu Val Cys Gly Glu Lys Ala Leu Lys Leu Gly Leu Gln Val Ile
275 280 285
Ala Lys Ile Thr Gly Tyr Ala Asp Ala Ala Gln Ala Pro Glu Leu Phe
290 295 300
Thr Thr Ser Pro Ala Leu Ala Ile Pro Lys Ala Ile Ser Asn Ala Gly
305 310 315 320
Ile Glu Ala Ser Gln Val Asp Tyr Tyr Glu Ile Asn Glu Ala Phe Ala
325 330 335
Val Val Ala Leu Ala Asn Gln Lys Leu Leu Gly Leu Ala Pro Glu Lys
340 345 350
Val Asn Val His Gly Gly Ala Val Ser Leu Gly His Pro Leu Gly Cys
355 360 365
Ser Gly Ala Arg Ile Val Val Thr Leu Leu Gly Val Leu Arg Gln Lys
370 375 380
Asn Gly Lys Tyr Gly Val Ser Gly Val Cys Asn Gly Gly Gly Gly Ala
385 390 395 400
Ser Ala Leu Val Val Glu Leu Val
405
<210>3
<211>1757
<212>DNA
<213>杜仲
<220>
<221>CDS
<222>(128)..(1354)
<223>乙酰-CoA C乙酰转移酶
<400>3
aatcactcct atagggaatt ccttaagatt tggacctctc tctctctctc tctctctctc 60
tctctccact gcttctggct gcttcactgt gagcaattaa ctgagatata catttgcata 120
tattatc atg gct cct gca gcg tca aca ggt ggt tca gag tca ata aag 169
Met Ala Pro Ala Ala Ser Thr Gly Gly Ser Glu Ser Ile Lys
1 5 10
cct aga gat gtt tgt att gtt ggt gtt gca cgt aca cca atg ggt ggc 217
Pro Arg Asp Val Cys Ile Val Gly Val Ala Arg Thr Pro Met Gly Gly
15 20 25 30
ttt ctt gga act ctt tcg tcc ttg gca gca acc aag ctt gga tct ata 265
Phe Leu Gly Thr Leu Ser Ser Leu Ala Ala Thr Lys Leu Gly Ser Ile
35 40 45
gct att gaa tgc gct ctt aaa aga gct ggc gtt gat cca tcg ctt gtg 313
Ala Ile Glu Cys Ala Leu Lys Arg Ala Gly Val Asp Pro Ser Leu Val
50 55 60
caa gaa gtg cta ttt ggg aat gtt ctc agt gca aat ttg ggg cag gct 361
Gln Glu Val Leu Phe Gly Asn Val Leu Ser Ala Asn Leu Gly Gln Ala
65 70 75
cct gct aga caa gca gca ctt ggt gca gga ata cct agc tca gtg gtt 409
Pro Ala Arg Gln Ala Ala Leu Gly Ala Gly Ile Pro Ser Ser Val Val
80 85 90
tgc acc aca att aac aaa gtt tgt gct tca gga atg aaa gca act atg 457
Cys Thr Thr Ile Asn Lys Val Cys Ala Ser Gly Met Lys Ala Thr Met
95 100 105 110
ctt gca gca cag agt atc cag ttg ggc ctc aat gac gtt gta gta gca 505
Leu Ala Ala Gln Ser Ile Gln Leu Gly Leu Asn Asp Val Val Val Ala
115 120 125
ggt ggc atg gaa agc atg tct aat gtc ccc aaa tac att gca gaa gca 553
Gly Gly Met Glu Ser Met Ser Asn Val Pro Lys Tyr Ile Ala Glu Ala
130 135 140
agg aag gga tct cgt ctt gga cac gat act ctt gtt gat gga atg ctg 601
Arg Lys Gly Ser Arg Leu Gly His Asp Thr Leu Val Asp Gly Met Leu
145 150 155
aaa gat gga ttg tgg gat gtt tat aat gat tgt gga atg gga gtt tgt 649
Lys Asp Gly Leu Trp Asp Val Tyr Asn Asp Cys Gly Met Gly Val Cys
160 165 170
gct gaa ata tgt gct gag cat cat gga gta aca aga gag gag cag gat 697
Ala Glu Ile Cys Ala Glu His His Gly Val Thr Arg Glu Glu Gln Asp
175 180 185 190
gat tat gct att cag agc ttc gaa cgt ggt att gcc gct caa gat agt 745
Asp Tyr Ala Ile Gln Ser Phe Glu Arg Gly Ile Ala Ala Gln Asp Ser
195 200 205
ggt gcc ttt gca tgg gag atc gtt cca gtt gaa gtt gct ggc gga agg 793
Gly Ala Phe Ala Trp Glu Ile Val Pro Val Glu Val Ala Gly Gly Arg
210 215 220
gga aaa cca tcg aca gtt gtg gat aaa gat gaa ggt tta gga aag ttt 841
Gly Lys Pro Ser Thr Val Val Asp Lys Asp Glu Gly Leu Gly Lys Phe
225 230 235
gat gct gca aag ttg agg aag cta cgg cca agt ttc aag gaa agt ggt 889
Asp Ala Ala Lys Leu Arg Lys Leu Arg Pro Ser Phe Lys Glu Ser Gly
240 245 250
ggt agt gtt act gct ggc aat gct tct agc ata agt gat ggt gct gcc 937
Gly Ser Val Thr Ala Gly Asn Ala Ser Ser Ile Ser Asp Gly Ala Ala
255 260 265 270
gca ctg gtt tta gtc agt gga cag aaa gca att gag ctt ggg ctg acc 985
Ala Leu Val Leu Val Ser Gly Gln Lys Ala Ile Glu Leu Gly Leu Thr
275 280 285
gtg att gca aaa atc aca ggt tat gct gat gct gct cag gca ccc gag 1033
Val Ile Ala Lys Ile Thr Gly Tyr Ala Asp Ala Ala Gln Ala Pro Glu
290 295 300
ttg ttt aca act tct ccg gcc ctt gca ata cca aaa gct att tca aat 1081
Leu Phe Thr Thr Ser Pro Ala Leu Ala Ile Pro Lys Ala Ile Ser Asn
305 310 315
gct ggc cta gag tcc tcc caa gtt gat tat tat gaa atc aat gaa gca 1129
Ala Gly Leu Glu Ser Ser Gln Val Asp Tyr Tyr Glu Ile Asn Glu Ala
320 325 330
ttc gcg gtt gtg gct ctt gca aat cag aaa ctg ctt gaa ctt gct cct 1177
Phe Ala Val Val Ala Leu Ala Asn Gln Lys Leu Leu Glu Leu Ala Pro
335 340 345 350
gac aaa gtt aac gta cat ggt gga gct gta tct cta ggg cat ccg cta 1225
Asp Lys Val Asn Val His Gly Gly Ala Val Ser Leu Gly His Pro Leu
355 360 365
ggt tgc agt ggt gcc cgt atc ttg gtc acc ctt ttg ggg gta tta aag 1273
Gly Cys Ser Gly Ala Arg Ile Leu Val Thr Leu Leu Gly Val Leu Lys
370 375 380
cag aag aat gcg aag tat gga gtt ggc ggt gtc tgt aac ggg gga ggt 1321
Gln Lys Asn Ala Lys Tyr Gly Val Gly Gly Val Cys Asn Gly Gly Gly
385 390 395
gga gca tca gcc ctc gtt gtt gag ctt ctg taa tcgcatttta ttagaatagt 1374
Gly Ala Ser Ala Leu Val Val Glu Leu Leu
400 405
taagaagaga aggatctaaa tctaactaag atttagcttg gccgtgtgtt caatgttgtt 1434
ctcttttttc tcctctctct tcttgattaa aaaaaaaaaa atgcaccctg gtgttacaaa 1494
gttccagcta tatctggagt ctgagtaaag atcggatcac ttgggtttat cgtacgcagc 1554
attagcttag tttgtaagat gttgtattga tgaatgaatt tgtatccaat aagaaggtgc 1614
atttttctaa tgatgtgatt ttgtgtttgg agagggcaat atgaaaagac gttgaggcca 1674
aaagttgtgg gtgttgaata aaccacatga atttgttgtt tttataaaag aaaccctatg 1734
taatttgcca aaaaaaaaaa aaa 1757
<210>4
<211>408
<212>PRT
<213>杜仲
<400>4
Met Ala Pro Ala Ala Ser Thr Gly Gly Ser Glu Ser Ile Lys Pro Arg
1 5 10 15
Asp Val Cys Ile Val Gly Val Ala Arg Thr Pro Met Gly Gly Phe Leu
20 25 30
Gly Thr Leu Ser Ser Leu Ala Ala Thr Lys Leu Gly Ser Ile Ala Ile
35 40 45
Glu Cys Ala Leu Lys Arg Ala Gly Val Asp Pro Ser Leu Val Gln Glu
50 55 60
Val Leu Phe Gly Asn Val Leu Ser Ala Asn Leu Gly Gln Ala Pro Ala
65 70 75 80
Arg Gln Ala Ala Leu Gly Ala Gly Ile Pro Ser Ser Val Val Cys Thr
85 90 95
Thr Ile Asn Lys Val Cys Ala Ser Gly Met Lys Ala Thr Met Leu Ala
100 105 110
Ala Gln Ser Ile Gln Leu Gly Leu Asn Asp Val Val Val Ala Gly Gly
115 120 125
Met Glu Ser Met Ser Asn Val Pro Lys Tyr Ile Ala Glu Ala Arg Lys
130 135 140
Gly Ser Arg Leu Gly His Asp Thr Leu Val Asp Gly Met Leu Lys Asp
145 150 155 160
Gly Leu Trp Asp Val Tyr Asn Asp Cys Gly Met Gly Val Cys Ala Glu
165 170 175
Ile Cys Ala Glu His His Gly Val Thr Arg Glu Glu Gln Asp Asp Tyr
180 185 190
Ala Ile Gln Ser Phe Glu Arg Gly Ile Ala Ala Gln Asp Ser Gly Ala
195 200 205
Phe Ala Trp Glu Ile Val Pro Val Glu Val Ala Gly Gly Arg Gly Lys
210 215 220
Pro Ser Thr Val Val Asp Lys Asp Glu Gly Leu Gly Lys Phe Asp Ala
225 230 235 240
Ala Lys Leu Arg Lys Leu Arg Pro Ser Phe Lys Glu Ser Gly Gly Ser
245 250 255
Val Thr Ala Gly Asn Ala Ser Ser Ile Ser Asp Gly Ala Ala Ala Leu
260 265 270
Val Leu Val Ser Gly Gln Lys Ala Ile Glu Leu Gly Leu Thr Val Ile
275 280 285
Ala Lys Ile Thr Gly Tyr Ala Asp Ala Ala Gln Ala Pro Glu Leu Phe
290 295 300
Thr Thr Ser Pro Ala Leu Ala Ile Pro Lys Ala Ile Ser Asn Ala Gly
305 310 315 320
Leu Glu Ser Ser Gln Val Asp Tyr Tyr Glu Ile Asn Glu Ala Phe Ala
325 330 335
Val Val Ala Leu Ala Asn Gln Lys Leu Leu Glu Leu Ala Pro Asp Lys
340 345 350
Val Asn Val His Gly Gly Ala Val Ser Leu Gly His Pro Leu Gly Cys
355 360 365
Ser Gly Ala Arg Ile Leu Val Thr Leu Leu Gly Val Leu Lys Gln Lys
370 375 380
Asn Ala Lys Tyr Gly Val Gly Gly Val Cys Asn Gly Gly Gly Gly Ala
385 390 395 400
Ser Ala Leu Val Val Glu Leu Leu
405
<210>5
<211>1892
<212>DNA
<213>杜仲
<220>
<221>CDS
<222>(172)..(1563)
<223>HMG-CoA合成酶
<400>5
aagatttgac tcctccgaat ttccagctct gtctctcctc tctctctctc catagctgtt 60
ctgctcagtg agggaagggt tggccgttgt tcttcctttg atctgtttgt tctcgagaat 120
ccgcgacgtt gaaggcgtgg gagggaggaa gaacgaacag gaagggcaat c atg gct 177
Met Ala
1
aaa aat gtt ggg att ctc gcg atg gaa atc tac ttc cct cct act tgt 225
Lys Asn Val Gly Ile Leu Ala Met Glu Ile Tyr Phe Pro Pro Thr Cys
5 10 15
att cag cag gaa aca ttg gaa gct cat gat gga gca agt aaa ggg aaa 273
Ile Gln Gln Glu Thr Leu Glu Ala His Asp Gly Ala Ser Lys Gly Lys
20 25 30
tac aca att gga ctt gga caa gat tgc atg gca ttt tgt acc gag gtt 321
Tyr Thr Ile Gly Leu Gly Gln Asp Cys Met Ala Phe Cys Thr Glu Val
35 40 45 50
gaa gat gtc atc tca atg agt ttg aca gcc gtt act tct ctt ctt gag 369
Glu Asp Val Ile Ser Met Ser Leu Thr Ala Val Thr Ser Leu Leu Glu
55 60 65
aaa tat ggg att gat cct aaa caa att ggt cgt ctt gaa gtg gga agt 417
Lys Tyr Gly Ile Asp Pro Lys Gln Ile Gly Arg Leu Glu Val Gly Ser
70 75 80
gag act gtt ata gac aag agc aaa tcc att aag aca ttc ctg atg caa 465
Glu Thr Val Ile Asp Lys Ser Lys Ser Ile Lys Thr Phe Leu Met Gln
85 90 95
atc ttt gag gaa tgt gga aat aca gac att gaa ggg gtt gac tca acc 513
Ile Phe Glu Glu Cys Gly Asn Thr Asp Ile Glu Gly Val Asp Ser Thr
100 105 110
aat gca tgc tat ggt gga act gct gca tta ttc aac tgt gtc aat tgg 561
Asn Ala Cys Tyr Gly Gly Thr Ala Ala Leu Phe Asn Cys Val Asn Trp
115 120 125 130
gtg gag agt agt tca tgg gat gga cgc tat gga ctt gtt gtg tgc acg 609
Val Glu Ser Ser Ser Trp Asp Gly Arg Tyr Gly Leu Val Val Cys Thr
135 140 145
gac agt gcg gtc tat gct gag gga cca gct cga cca act ggt ggt gcc 657
Asp Ser Ala Val Tyr Ala Glu Gly Pro Ala Arg Pro Thr Gly Gly Ala
150 155 160
gct gct att gct atc cta ata ggg cct gat gct cct att gca ttt gag 705
Ala Ala Ile Ala Ile Leu Ile Gly Pro Asp Ala Pro Ile Ala Phe Glu
165 170 175
agc aaa atc agg gga agc cat atg tct cac gcc tat gac ttc tac aag 753
Ser Lys Ile Arg Gly Ser His Met Ser His Ala Tyr Asp Phe Tyr Lys
180 185 190
ccc aac cta gca agt gaa tat ccg gtt gtt gat gga aaa ctt tca caa 801
Pro Asn Leu Ala Ser Glu Tyr Pro Val Val Asp Gly Lys Leu Ser Gln
195 200 205 210
acg tgt tat ctt atg gca ctt gat tct tgc tac aaa cgc ctc tgt cat 849
Thr Cys Tyr Leu Met Ala Leu Asp Ser Cys Tyr Lys Arg Leu Cys His
215 220 225
aag tac gag aaa ctg gag ggt aaa caa ttt tct gta aat gat gcc gac 897
Lys Tyr Glu Lys Leu Glu Gly Lys Gln Phe Ser Val Asn Asp Ala Asp
230 235 240
tac ttt gta ttt cat tct ccg tat aac aag ctt gta cag aag agc ttt 945
Tyr Phe Val Phe His Ser Pro Tyr Asn Lys Leu Val Gln Lys Ser Phe
245 250 255
gct cga ttg ttg ttc aac gat tac ttg agg aac gcc aac tcc att gat 993
Ala Arg Leu Leu Phe Asn Asp Tyr Leu Arg Asn Ala Asn Ser Ile Asp
260 265 270
ggg gca gca aag gaa aag ctg gga ccc ttt tct tcc tta act ggt gat 1041
Gly Ala Ala Lys Glu Lys Leu Gly Pro Phe Ser Ser Leu Thr Gly Asp
275 280 285 290
gaa agc tac cag agt cgt gat ctt gag aag gct acc caa caa gtt gca 1089
Glu Ser Tyr Gln Ser Arg Asp Leu Glu Lys Ala Thr Gln Gln Val Ala
295 300 305
aaa ccc ttt tat gat gca aaa gtt caa cca aca act tta ata cca aag 1137
Lys Pro Phe Tyr Asp Ala Lys Val Gln Pro Thr Thr LeuIle Pro Lys
310 315 320
caa gtc gga aac atg tat acc gca tct cta tat act gca ttt gca tcc 1185
Gln Val Gly Asn Met Tyr Thr Ala Ser Leu Tyr Thr Ala Phe Ala Ser
325 330 335
ctt att cac aac aaa cac gat aca ctg gat ggg cag agg gtg ata ttg 1233
Leu Ile His Asn Lys His Asp Thr Leu Asp Gly Gln Arg Val Ile Leu
340 345 350
ttc tct tat ggg agt gga tta acc gcc acg atg ttt tct ctc cgg att 1281
Phe Ser Tyr Gly Ser Gly Leu Thr Ala Thr Met Phe Ser Leu Arg Ile
355 360 365 370
cgc agt aag cca caa cct ttt tgc ttg tca aac atc gtg aat gtg atg 1329
Arg Ser Lys Pro Gln Pro Phe Cys Leu Ser Asn Ile Val Asn Val Met
375 380 385
aat gtt gct gag aaa ttg aat tcg aga cat gag gtc tct cca gaa aaa 1377
Asn Val Ala Glu Lys Leu Asn Ser Arg His Glu Val Ser Pro Glu Lys
390 395 400
ttc gtc gaa aca atg aag cta atg gag cat agg tat gga gcc aag gac 1425
Phe Val Glu Thr Met Lys Leu Met Glu His Arg Tyr Gly Ala Lys Asp
405 410 415
ttt gta act agc aaa gac att agc ctt cta tct cca ggc aca tac tat 1473
Phe Val Thr Ser Lys Asp Ile Ser Leu Leu Ser Pro Gly Thr Tyr Tyr
420 425 430
ctc acc cat gtg gac tct atg tac cga aga ttc tat gag aag gcg gca 1521
Leu Thr His Val Asp Ser Met Tyr Arg Arg Phe Tyr Glu Lys Ala Ala
435 440 445 450
aaa acc gct cat gag aac ggc tca ttg gct aac ggt cac taa 1563
Lys Thr Ala His Glu Asn Gly Ser Leu Ala Asn Gly His
455 460
taacaataga ctgaccaaaa ctggcattta tttttctagc atcatagaac aataatttgg 1623
tttgcgaatc agtttgttct gctttttagc tactcgtttg tgtgtcacgg ggaaatcccc 1683
attggcactc ctgaaatgat cacctgcact agaaatcatg tacttggagt gcaaatgaac 1743
atttcgggag agtcaatagt gttttaaggc atgtaaaagg caattctata aaggttgtaa 1803
tttcttcgtt gaagttgaaa cgttatattg tgtttataac tatgaaaata ttgatatttt 1863
ctatattttc atgaaaaaaa aaaaaaaaa 1892
<210>6
<211>463
<212>PRT
<213>杜仲
<400>6
Met Ala Lys Asn Val Gly Ile Leu Ala Met Glu Ile Tyr Phe Pro Pro
1 5 10 15
Thr Cys Ile Gln Gln Glu Thr Leu Glu Ala His Asp Gly Ala Ser Lys
20 25 30
Gly Lys Tyr Thr Ile Gly Leu Gly Gln Asp Cys Met Ala Phe Cys Thr
35 40 45
Glu Val Glu Asp Val Ile Ser Met Ser Leu Thr Ala Val Thr Ser Leu
50 55 60
Leu Glu Lys Tyr Gly Ile Asp Pro Lys Gln Ile Gly Arg Leu Glu Val
65 70 75 80
Gly Ser Glu Thr Val Ile Asp Lys Ser Lys Ser Ile Lys Thr Phe Leu
85 90 95
Met Gln Ile Phe Glu Glu Cys Gly Asn Thr Asp Ile Glu Gly Val Asp
100 105 110
Ser Thr Asn Ala Cys Tyr Gly Gly Thr Ala Ala Leu Phe Asn Cys Val
115 120 125
Asn Trp Val Glu Ser Ser Ser Trp Asp Gly Arg Tyr Gly Leu Val Val
130 135 140
Cys Thr Asp Ser Ala Val Tyr Ala Glu Gly Pro Ala Arg Pro Thr Gly
145 150 155 160
Gly Ala Ala Ala Ile Ala Ile Leu Ile Gly Pro Asp Ala Pro Ile Ala
165 170 175
Phe Glu Ser Lys Ile Arg Gly Ser His Met Ser His Ala Tyr Asp Phe
180 185 190
Tyr Lys Pro Asn Leu Ala Ser Glu Tyr Pro Val Val Asp Gly Lys Leu
195 200 205
Ser Gln Thr Cys Tyr Leu Met Ala Leu Asp Ser Cys Tyr Lys Arg Leu
210 215 220
Cys His Lys Tyr Glu Lys Leu Glu Gly Lys Gln Phe Ser Val Asn Asp
225 230 235 240
Ala Asp Tyr Phe Val Phe His Ser Pro Tyr Asn Lys Leu Val Gln Lys
245 250 255
Ser Phe Ala Arg Leu Leu Phe Asn Asp Tyr Leu Arg Asn Ala Asn Ser
260 265 270
Ile Asp Gly Ala Ala Lys Glu Lys Leu Gly Pro Phe Ser Ser Leu Thr
275 280 285
Gly Asp Glu Ser Tyr Gln Ser Arg Asp Leu Glu Lys Ala Thr Gln Gln
290 295 300
Val Ala Lys Pro Phe Tyr Asp Ala Lys Val Gln Pro Thr Thr Leu Ile
305 310 315 320
Pro Lys Gln Val Gly Asn Met Tyr Thr Ala Ser Leu Tyr Thr Ala Phe
325 330 335
Ala Ser Leu Ile His Asn Lys His Asp Thr Leu Asp Gly Gln Arg Val
340 345 350
Ile Leu Phe Ser Tyr Gly Ser Gly Leu Thr Ala Thr Met Phe Ser Leu
355 360 365
Arg Ile Arg Ser Lys Pro Gln Pro Phe Cys Leu Ser Asn Ile Val Asn
370 375 380
Val Met Asn Val Ala Glu Lys Leu Asn Ser Arg His Glu Val Ser Pro
385 390 395 400
Glu Lys Phe Val Glu Thr Met Lys Leu Met Glu His Arg Tyr Gly Ala
405 410 415
Lys Asp Phe Val Thr Ser Lys Asp Ile Ser Leu Leu Ser Pro Gly Thr
420 425 430
Tyr Tyr Leu Thr His Val Asp Ser Met Tyr Arg Arg Phe Tyr Glu Lys
435 440 445
Ala Ala Lys Thr Ala His Glu Asn Gly Ser Leu Ala Asn Gly His
450 455 460
<210>7
<211>1833
<212>DNA
<213>杜仲
<220>
<221>CDS
<222>(216)..(1616)
<223>HMG-CoA合成酶
<400>7
cgttttatcg actcctatag ggaattcctt aagatttgag caactttcca gtttccagtt 60
tccagactcc cccccccccc cctctctctc tctctctctc tctctctccg tctgcgaaaa 120
tcaaaattta tagagagaag aattgttctt catcaatttc atccgttcgt ttctcgtgag 180
gtttaaagga gagggaaaga gagaagaagg aaaaa atg gct tcg caa ccg aaa 233
Met Ala Ser Gln Pro Lys
1 5
aat gtt gga att ctc gct atg gag atc tat ttt cct ccc acc tgt gtc 281
Asn Val Gly Ile Leu Ala Met Glu Ile Tyr Phe Pro Pro Thr Cys Val
10 15 20
cag cag gaa gca atg gaa act cat gat gga gta agt aaa gga aag tac 329
Gln Gln Glu Ala Met Glu Thr His Asp Gly Val Ser Lys Gly Lys Tyr
25 30 35
aca att ggc ctt gga caa gat tgc atg gcg ttt tgt acg gag gtc gaa 377
Thr Ile Gly Leu Gly Gln Asp Cys Met Ala Phe Cys Thr Glu Val Glu
40 45 50
gat gtc att tcc atg agt ttg aca gct gtt act tcc ctt ctt gag aaa 425
Asp Val Ile Ser Met Ser Leu Thr Ala Val Thr Ser Leu Leu Glu Lys
55 60 65 70
tac ggg atc gat ccc acg caa att ggt cgt atg gaa gtg ggc agt gaa 473
Tyr Gly Ile Asp Pro Thr Gln Ile Gly Arg Met Glu Val Gly Ser Glu
75 80 85
act gtt att gac aag agc aaa tcc att aaa act ttc ttg atg caa att 521
Thr Val Ile Asp Lys Ser Lys Ser Ile Lys Thr Phe Leu Met Gln Ile
90 95 100
ttt gag gaa tgt gga aat acc gac att gaa ggt gtt gat tca acc aat 569
Phe Glu Glu Cys Gly Asn Thr AspIle Glu Gly Val Asp Ser Thr Asn
105 110 115
gct tgc tat ggg gga acc gct gca tta cta aat tgc cca att ggg tgg 617
Ala Cys Tyr Gly Gly Thr Ala Ala Leu Leu Asn Cys Pro Ile Gly Trp
120 125 130
aga gca gtt cgt ggg atg gac gtt atg gaa ctt gtt gtg tgc acc gat 665
Arg Ala Val Arg Gly Met Asp Val Met Glu Leu Val Val Cys Thr Asp
135 140 145 150
agt gcg gtc tat gct gaa gga cca gct cga ccg act ggt gga gct gct 713
Ser Ala Val Tyr Ala Glu Gly Pro Ala Arg Pro Thr Gly Gly Ala Ala
155 160 165
gct att gcc atg ctg att ggc ccc gat gct cca att gca ttt gaa agc 761
Ala Ile Ala Met Leu Ile Gly Pro Asp Ala Pro Ile Ala Phe Glu Ser
170 175 180
aaa atc agg gcg agt cat atg tct cat gcc tat gat ttc tac aag cca 809
Lys Ile Arg Ala Ser His Met Ser His Ala Tyr Asp Phe Tyr Lys Pro
185 190 195
aat ctt gca agt gaa tat ccg gtt gtt gat gga aag ctt tcc caa act 857
Asn Leu Ala Ser Glu Tyr Pro Val Val Asp Gly Lys Leu Ser Gln Thr
200 205 210
tgt tat ctg atg gca ctt gat tct tgc tac aac cgt tta tgc aat aag 905
Cys Tyr Leu Met Ala Leu Asp Ser Cys Tyr Asn Arg Leu Cys Asn Lys
215 220 225 230
tac aag aaa ttg gag ggc aaa caa ttt tcg gtt gcc gat gct gac tac 953
Tyr Lys Lys Leu Glu Gly Lys Gln Phe Ser Val Ala Asp Ala Asp Tyr
235 240 245
ttt gta ttt cac tct cca tac aac aag ctt gta cag aag agc ttt gct 1001
Phe Val Phe His Ser Pro Tyr Asn Lys Leu Val Gln Lys Ser Phe Ala
250 255 260
cga ttg ttg ttc aat gat ttc ttg aga aat gcc agc tct att gat gag 1049
Arg Leu Leu Phe Asn Asp Phe Leu Arg Asn Ala Ser Ser Ile Asp Glu
265 270 275
gct gct aaa gaa aag ctc agt ccg ttt tca tct tta acc ggc gat gaa 1097
Ala Ala Lys Glu Lys Leu Ser Pro Phe Ser Ser Leu Thr Gly Asp Glu
280 285 290
agc tac caa agc cgt gat ctt gaa aag gcg tcc cag caa gtg gca aag 1145
Ser Tyr Gln Ser Arg Asp Leu Glu Lys Ala Ser Gln Gln Val Ala Lys
295 300 305 310
cca ttc tat gat gaa aaa gtt caa ccc tcc act ttg ata cca aag caa 1193
Pro Phe Tyr Asp Glu Lys Val Gln Pro Ser Thr Leu Ile Pro Lys Gln
315 320 325
gtc gga aac atg tac act gcc tct cta tat gct gca ttt gcc tcc ctt 1241
Val Gly Asn Met Tyr Thr Ala Ser Leu Tyr Ala Ala Phe Ala Ser Leu
330 335 340
ctt cac aac aag cat agc aca ctg gct ggg cag agg gtg ata att ttc 1289
Leu His Asn Lys His Ser Thr Leu Ala Gly Gln Arg Val Ile Ile Phe
345 350 355
tct tac ggg agt gga ttg acg gcc acg atg ttc tct ttc cgt att cgc 1337
Ser Tyr Gly Ser Gly Leu Thr Ala Thr Met Phe Ser Phe Arg Ile Arg
360 365 370
gaa ggc caa cac cct ttt agc ttg gga aac att gcc aaa gtg atg aat 1385
Glu Gly Gln His Pro Phe Ser Leu Gly Asn Ile Ala Lys Val Met Asn
375 380 385 390
gtt gac gaa aaa ttg aag tcc agg cat gag ttt acg cct gaa aaa ttc 1433
Val Asp Glu Lys Leu Lys Ser Arg His Glu Phe Thr Pro Glu Lys Phe
395 400 405
gtc gaa act atg aag cta atg gag cat agg tac gga gca aag gac att 1481
Val Glu Thr Met Lys Leu Met Glu His Arg Tyr Gly Ala Lys Asp Ile
410 415 420
gtg aca agc aag gat aca agc ctt ctt tca cca ggc act tac tat ctc 1529
Val Thr Ser Lys Asp Thr Ser Leu Leu Ser Pro Gly Thr Tyr Tyr Leu
425 430 435
act gaa gtc gac tcg atg tat cga aga ttc tac tcc aag aaa gat gaa 1577
Thr Glu Val Asp Ser Met Tyr Arg Arg Phe Tyr Ser Lys Lys Asp Glu
440 445 450
aac aca agc gag ggc tta ccg cat tca aat ggt ttt tga attcacgaaa 1626
Asn Thr Ser Glu Gly Leu Pro His Ser Asn Gly Phe
455 460 465
tttagcgatt caaagatgtg atttttttta agaataacgg tgattctggg tagaaaatat 1686
agttttgcct gggagaatct gcctccatgt ttattttagt tgaattcaac tttcaagcaa 1746
atgtttgtca atggttgatc attgattgtt tgatgtattg tgggtttgcc taatgaaatg 1806
tctacgttgc tactaaaaaa aaaaaaa 1833
<210>8
<211>466
<212>PRT
<213>杜仲
<400>8
Met Ala Ser Gln Pro Lys Asn Val Gly Ile Leu Ala Met Glu Ile Tyr
1 5 10 15
Phe Pro Pro Thr Cys Val Gln Gln Glu Ala Met Glu Thr His Asp Gly
20 25 30
Val Ser Lys Gly Lys Tyr Thr Ile Gly Leu Gly Gln Asp Cys Met Ala
35 40 45
Phe Cys Thr Glu Val Glu Asp Val Ile Ser Met Ser Leu Thr Ala Val
50 55 60
Thr Ser Leu Leu Glu Lys Tyr Gly Ile Asp Pro Thr Gln Ile Gly Arg
65 70 75 80
Met Glu Val Gly Ser Glu Thr Val Ile Asp Lys Ser Lys Ser Ile Lys
85 90 95
Thr Phe Leu Met Gln Ile Phe Glu Glu Cys Gly Asn Thr Asp Ile Glu
100 105 110
Gly Val Asp Ser Thr Asn Ala Cys Tyr Gly Gly Thr Ala Ala Leu Leu
115 120 125
Asn Cys Pro Ile Gly Trp Arg Ala Val Arg Gly Met Asp Val Met Glu
130 135 140
Leu Val Val Cys Thr Asp Ser Ala Val Tyr Ala Glu Gly Pro Ala Arg
145 150 155 160
Pro Thr Gly Gly Ala Ala Ala Ile Ala Met Leu Ile Gly Pro Asp Ala
165 170 175
Pro Ile Ala Phe Glu Ser Lys Ile Arg Ala Ser His Met Ser His Ala
180 185 190
Tyr Asp Phe Tyr Lys Pro Asn Leu Ala Ser Glu Tyr Pro Val Val Asp
195 200 205
Gly Lys Leu Ser Gln Thr Cys Tyr Leu Met Ala Leu Asp Ser Cys Tyr
210 215 220
Asn Arg Leu Cys Asn Lys Tyr Lys Lys Leu Glu Gly Lys Gln Phe Ser
225 230 235 240
Val Ala Asp Ala Asp Tyr Phe Val Phe His Ser Pro Tyr Asn Lys Leu
245 250 255
Val Gln Lys Ser Phe Ala Arg Leu Leu Phe Asn Asp Phe Leu Arg Asn
260 265 270
Ala Ser Ser Ile Asp Glu Ala Ala Lys Glu Lys Leu Ser Pro Phe Ser
275 280 285
Ser Leu Thr Gly Asp Glu Ser Tyr Gln Ser Arg Asp Leu Glu Lys Ala
290 295 300
Ser Gln Gln Val Ala Lys Pro Phe Tyr Asp Glu Lys Val Gln Pro Ser
305 310 315 320
Thr Leu Ile Pro Lys Gln Val Gly Asn Met Tyr Thr Ala Ser Leu Tyr
325 330 335
Ala Ala Phe Ala Ser Leu Leu His Asn Lys His Ser Thr Leu Ala Gly
340 345 350
Gln Arg Val Ile Ile Phe Ser Tyr Gly Ser Gly Leu Thr Ala Thr Met
355 360 365
Phe Ser Phe Arg Ile Arg Glu Gly Gln His Pro Phe Ser Leu Gly Asn
370 375 380
Ile Ala Lys Val Met Asn Val Asp Glu Lys Leu Lys Ser Arg His Glu
385 390 395 400
Phe Thr Pro Glu Lys Phe Val Glu Thr Met Lys Leu Met Glu His Arg
405 410 415
Tyr Gly Ala Lys Asp Ile Val Thr Ser Lys Asp Thr Ser Leu Leu Ser
420 425 430
Pro Gly Thr Tyr Tyr Leu Thr Glu Val Asp Ser Met Tyr Arg Arg Phe
435 440 445
Tyr Ser Lys Lys Asp Glu Asn Thr Ser Glu Gly Leu Pro His Ser Asn
450 455 460
Gly Phe
465
<210>9
<211>1825
<212>DNA
<213>杜仲
<220>
<221>CDS
<222>(216)..(1616)
<223>HMG-CoA合成酶
<400>9
cgttttatcg actcctatag ggaattcctt aagatttgag caactttcca gtttccagtt 60
tccagactcc cccccccccc cctctctctc tctctctctc tctctctccg tctgcgaaaa 120
tcaaaattta tagagagaag aattgttctt catcaatttc atccgttcgt ttctcgtgag 180
gtttaaagga gagggaaaga gagaagaagg aaaaa atg gct tcg caa ccg aaa 233
Met Ala Ser Gln Pro Lys
1 5
aat gtt gga att ctc gct atg gag atc tat ttt cct ccc acc tgt gtc 281
Asn Val Gly Ile Leu Ala Met Glu Ile Tyr Phe Pro Pro Thr Cys Val
10 15 20
cag cag gaa gca atg gaa act cat gat gga gta agt aaa gga aag tac 329
Gln Gln Glu Ala Met Glu Thr His Asp Gly Val Ser Lys Gly Lys Tyr
25 30 35
aca att ggc ctt gga caa gat tgc atg gcg ttt tgt acg gag gtc gaa 377
Thr Ile Gly Leu Gly Gln Asp Cys Met Ala Phe Cys Thr Glu Val Glu
40 45 50
gat gtc att tcc atg agt ttg aca gct gtt act tcc ctt ctt gag aaa 425
Asp Val Ile Ser Met Ser Leu Thr Ala Val Thr Ser Leu Leu Glu Lys
55 60 65 70
tac ggg atc gat ccc acg caa att ggt cgt atg gaa gtg ggc agt gaa 473
Tyr Gly Ile Asp Pro Thr Gln Ile Gly Arg Met Glu Val Gly Ser Glu
75 80 85
act gtt att gac aag agc aaa tcc att aaa act ttc ttg atg caa att 521
Thr Val Ile Asp Lys Ser Lys Ser Ile Lys Thr Phe Leu Met Gln Ile
90 95 100
ttt gag gaa tgt gga aat acc gac att gaa ggt gtt gat tca acc aat 569
Phe Glu Glu Cys Gly Asn Thr Asp Ile Glu Gly Val Asp Ser Thr Asn
105 110 115
gct tgc tat ggg gga acc gct gca tta cta aat tgc cca att ggg tgg 617
Ala Cys Tyr Gly Gly Thr Ala Ala Leu Leu Asn Cys Pro Ile Gly Trp
120 125 130
aga gca gtt cgt ggg atg gac gtt atg gaa ctt gtt gtg tgc acc gat 665
Arg Ala Val Arg Gly Met Asp Val Met Glu Leu Val Val Cys Thr Asp
135 140 145 150
agt gcg gtc tat gct gaa gga cca gct cga ccg act ggt gga gct gct 713
Ser Ala Val Tyr Ala Glu Gly Pro Ala Arg Pro Thr Gly Gly Ala Ala
155 160 165
gct att gcc atg ctg att ggc ccc gat gct cca att gca ttt gaa agc 761
Ala Ile Ala Met Leu Ile Gly Pro Asp Ala Pro Ile Ala Phe Glu Ser
170 175 180
aaa atc agg gcg agt cat atg tct cat gcc tat gat ttc tac aag cca 809
Lys Ile Arg Ala Ser His Met Ser His Ala Tyr Asp Phe Tyr Lys Pro
185 190 195
aat ctt gca agt gaa tat ccg gtt gtt gat gga aag ctt tcc caa act 857
Asn Leu Ala Ser Glu Tyr Pro Val Val Asp Gly Lys Leu Ser Gln Thr
200 205 210
tgt tat ctg atg gca ctt gat tct tgc tac aac cgt tta tgc aat aag 905
Cys Tyr Leu Met Ala Leu Asp Ser Cys Tyr Asn Arg Leu Cys Asn Lys
215 220 225 230
tac aag aaa ttg gag ggc aaa caa ttt tcg gtt gcc gat gct gac tac 953
Tyr Lys Lys Leu Glu Gly Lys Gln Phe Ser Val Ala Asp Ala Asp Tyr
235 240 245
ttt gta ttt cac tct cca tac aac aag ctt gta cag aag agc ttt gct 1001
Phe Val Phe His Ser Pro Tyr Asn Lys Leu Val Gln Lys Ser Phe Ala
250 255 260
cga ttg ttg ttc aat gat ttc ttg aga aat gcc agc tct att gat gag 1049
Arg Leu Leu Phe Asn Asp Phe Leu Arg Asn Ala Ser Ser Ile Asp Glu
265 270 275
gct gct aaa gaa aag ctc agt ccg ttt tca tct tta acc ggc gat gaa 1097
Ala Ala Lys Glu Lys Leu Ser Pro Phe Ser Ser Leu Thr Gly Asp Glu
280 285 290
agc tac caa agc cgt gat ctt gaa aag gcg tcc cag caa gtg gca aag 1145
Ser Tyr Gln Ser Arg Asp Leu Glu Lys Ala Ser Gln Gln Val Ala Lys
295 300 305 310
cca ttc tat gat gaa aaa gtt caa ccc tcc act ttg ata cca aag caa 1193
Pro Phe Tyr Asp Glu Lys Val Gln Pro Ser Thr Leu Ile Pro Lys Gln
315 320 325
gtc gga aac atg tac act gcc tct cta tat gct gca ttt gcc tcc ctt 1241
Val Gly Asn Met Tyr Thr Ala Ser Leu Tyr Ala Ala Phe Ala Ser Leu
330 335 340
ctt cac aac aag cat agc aca ctg gct ggg cag agg gtg ata att ttc 1289
Leu His Asn Lys His Ser Thr Leu Ala Gly Gln Arg Val Ile Ile Phe
345 350 355
tct tac ggg agt gga ttg acg gcc acg atg ttc tct ttc cgt att cgc 1337
Ser Tyr Gly Ser Gly Leu Thr Ala Thr Met Phe Ser Phe Arg Ile Arg
360 365 370
gaa ggc caa cac cct ttt agc ttg gga aac att gcc aaa gtg atg aat 1385
Glu Gly Gln His Pro Phe Ser Leu Gly Asn Ile Ala Lys Val Met Asn
375 380 385 390
gtt gac gaa aaa ttg aag tcc agg cat gag ttt acg cct gaa aaa ttc 1433
Val Asp Glu Lys Leu Lys Ser Arg His Glu Phe Thr Pro Glu Lys Phe
395 400 405
gtc gaa act atg aag cta atg gag cat agg tac gga gca aag gac att 1481
Val Glu Thr Met Lys Leu Met Glu His Arg Tyr Gly Ala Lys Asp Ile
410 415 420
gtg aca agc aag gat aca agc ctt ctt tca cca ggc act tac tat ctc 1529
Val Thr Ser Lys Asp Thr Ser Leu Leu Ser Pro Gly Thr Tyr Tyr Leu
425 430 435
act gaa gtc gac tcg atg tat cga aga ttc tac tcc aag aaa gat gaa 1577
Thr Glu Val Asp Ser Met Tyr Arg Arg Phe Tyr Ser Lys Lys Asp Glu
440 445 450
aac aca agc gag ggc tta ccg cat tca aat ggt ttt tga attcacgaaa 1626
Asn Thr Ser Glu Gly Leu Pro His Ser Asn Gly Phe
455 460 465
tttagcgatt caaagatgtg atttttttta agaataacgg tgattctggg tagaaaatat 1686
aggtttgcct gggagaatct gcatccgtgt ttattttagt tgaattcaac tttcaagcga 1746
atgtttgtcg atggttgatc attgattgtt gatgtattgt gggtttgcct aatgaaatgt 1806
ctacgttgct acaaaaaaa 1825
<210>10
<211>466
<212>PRT
<213>杜仲
<400>10
Met Ala Ser Gln Pro Lys Asn Val Gly Ile Leu Ala Met Glu Ile Tyr
1 5 10 15
Phe Pro Pro Thr Cys Val Gln Gln Glu Ala Met Glu Thr His Asp Gly
20 25 30
Val Ser Lys Gly Lys Tyr Thr Ile Gly Leu Gly Gln Asp Cys Met Ala
35 40 45
Phe Cys Thr Glu Val Glu Asp Val Ile Ser Met Ser Leu Thr Ala Val
50 55 60
Thr Ser Leu Leu Glu Lys Tyr Gly Ile Asp Pro Thr Gln Ile Gly Arg
65 70 75 80
Met Glu Val Gly Ser Glu Thr Val Ile Asp Lys Ser Lys Ser Ile Lys
85 90 95
Thr Phe Leu Met Gln Ile Phe Glu Glu Cys Gly Asn Thr Asp Ile Glu
100 105 110
Gly Val Asp Ser Thr Asn Ala Cys Tyr Gly Gly Thr Ala Ala Leu Leu
115 120 125
Asn Cys Pro Ile Gly Trp Arg Ala Val Arg Gly Met Asp Val Met Glu
130 135 140
Leu Val Val Cys Thr Asp Ser Ala Val Tyr Ala Glu Gly Pro Ala Arg
145 150 155 160
Pro Thr Gly Gly Ala Ala Ala Ile Ala Met Leu Ile Gly Pro Asp Ala
165 170 175
Pro Ile Ala Phe Glu Ser Lys Ile Arg Ala Ser His Met Ser His Ala
180 185 190
Tyr Asp Phe Tyr Lys Pro Asn Leu Ala Ser Glu Tyr Pro Val Val Asp
195 200 205
Gly Lys Leu Ser Gln Thr Cys Tyr Leu Met Ala Leu Asp Ser Cys Tyr
210 215 220
Asn Arg Leu Cys Asn Lys Tyr Lys Lys Leu Glu Gly Lys Gln Phe Ser
225 230 235 240
Val Ala Asp Ala Asp Tyr Phe Val Phe His Ser Pro Tyr Asn Lys Leu
245 250 255
Val Gln Lys Ser Phe Ala Arg Leu Leu Phe Asn Asp Phe Leu Arg Asn
260 265 270
Ala Ser Ser Ile Asp Glu Ala Ala Lys Glu Lys Leu Ser Pro Phe Ser
275 280 285
Ser Leu Thr Gly Asp Glu Ser Tyr Gln Ser Arg Asp Leu Glu Lys Ala
290 295 300
Ser Gln Gln Val Ala Lys Pro Phe Tyr Asp Glu Lys Val Gln Pro Ser
305 310 315 320
Thr Leu Ile Pro Lys Gln Val Gly Asn Met Tyr Thr Ala Ser Leu Tyr
325 330 335
Ala Ala Phe Ala Ser Leu Leu His Asn Lys His Ser Thr Leu Ala Gly
340 345 350
Gln Arg Val Ile Ile Phe Ser Tyr Gly Ser Gly Leu Thr Ala Thr Met
355 360 365
Phe Ser Phe Arg Ile Arg Glu Gly Gln His Pro Phe Ser Leu Gly Asn
370 375 380
Ile Ala Lys Val Met Asn Val Asp Glu Lys Leu Lys Ser Arg His Glu
385 390 395 400
Phe Thr Pro Glu Lys Phe Val Glu Thr Met Lys Leu Met Glu His Arg
405 410 415
Tyr Gly Ala Lys Asp Ile Val Thr Ser Lys Asp Thr Ser Leu Leu Ser
420 425 430
Pro Gly Thr Tyr Tyr Leu Thr Glu Val Asp Ser Met Tyr Arg Arg Phe
435 440 445
Tyr Ser Lys Lys Asp Glu Asn Thr Ser Glu Gly Leu Pro His Ser Asn
450 455 460
Gly Phe
465
<210>11
<211>2057
<212>DNA
<213>杜仲
<220>
<221>CDS
<222>(32)..(1921)
<223>HMG-CoA还原酶
<400>11
atcactccta tagggaatcc ttaagattta a atg tgg aac tca aac aaa aat 52
Met Trp Asn Ser Asn Lys Asn
1 5
cac ata cgc cac tct ctc tct cct tcc cgt ttc tca aga tac aca tac 100
His Ile Arg His Ser Leu Ser Pro Ser Arg Phe Ser Arg Tyr Thr Tyr
10 15 20
atc gat tcg tcg acg gaa tcg aaa tca atg gac gtc cga cgg cga tcg 148
Ile Asp Ser Ser Thr Glu Ser Lys Ser Met Asp Val Arg Arg Arg Ser
25 30 35
atg aaa ccc acc gcc gtt cat cat acc ccc acg cat tca tat gcc ctc 196
Met Lys Pro Thr Ala Val His His Thr Pro Thr His Ser Tyr Ala Leu
40 45 50 55
aaa ccc ccc caa aag gat gac gat gat tta cct aaa gct tcc gat gca 244
Lys Pro Pro Gln Lys Asp Asp Asp Asp Leu Pro Lys Ala Ser Asp Ala
60 65 70
ctt cct ttc cat ctt tac cta aca aac gct gta ttc ttc acc ctc ttc 292
Leu Pro Phe His Leu Tyr Leu Thr Asn Ala Val Phe Phe Thr Leu Phe
75 80 85
ttc tcc gtc gtg tat tac ctc ctc agc agg tgg cgc gaa aag atc cgc 340
Phe Ser Val Val Tyr Tyr Leu Leu Ser Arg Trp Arg Glu Lys Ile Arg
90 95 100
aac tcc act cct ctc cac atc gta tct ctc tcc gag atc gcg gca ata 388
Asn Ser Thr Pro Leu His Ile Val Ser Leu Ser Glu Ile Ala Ala Ile
105 110 115
gtc gcc ttc gtt gct tcc ttc att tac ctc ctt ggg ttc ttc ggc atc 436
Val Ala Phe Val Ala Ser Phe Ile Tyr Leu Leu Gly Phe Phe Gly Ile
120 125 130 135
gga ttc gtt cag tcc ttc gct ccc cgc tct tcc cac gac cat gat gtc 484
Gly Phe Val Gln Ser Phe Ala Pro Arg Ser Ser His Asp His Asp Val
140 145 150
gac gac gaa att tgg gtt aac gat gac atc atc cgt aag gat gac gac 532
Asp Asp Glu Ile Trp Val Asn Asp Asp Ile Ile Arg Lys Asp Asp Asp
155 160 165
ggt cga acc ggc cct tgt gct gtc aat tgc cca ctt ccc aaa ccg tac 580
Gly Arg Thr Gly Pro Cys Ala Val Asn Cys Pro Leu Pro Lys Pro Tyr
170 175 180
aac aag gtg gtt gta gaa gaa tcc ccg gat cca cct ctc gtc tca tct 628
Asn Lys Val Val Val Glu Glu Ser Pro Asp Pro Pro Leu Val Ser Ser
185 190 195
gca gag gac gag gaa atc atc aat tct gtc gtc gaa ggg aag att ccg 676
Ala Glu Asp Glu Glu Ile Ile Asn Ser Val Val Glu Gly Lys Ile Pro
200 205 210 215
tcc tat gct ctc gaa tcg aag ctt agt gat tgc cgt cgt gcc gca gca 724
Ser Tyr Ala Leu Glu Ser Lys Leu Ser Asp Cys Arg Arg Ala Ala Ala
220 225 230
gtt cgc cgc gaa gca ttg caa agg atg acc ggg aag tcc ctc gtg ggg 772
Val Arg Arg Glu Ala Leu Gln Arg Met Thr Gly Lys Ser Leu Val Gly
235 240 245
ctg cca ctg gag ggc ttc gat tac gag tcc att ctg ggg cag tgt tgc 820
Leu Pro Leu Glu Gly Phe Asp Tyr Glu Ser Ile Leu Gly Gln Cys Cys
250 255 260
gag atg cca gtg ggg tac gtc cag atc cca gtg ggt att gca ggg ccg 868
Glu Met Pro Val Gly Tyr Val Gln Ile Pro Val Gly Ile Ala Gly Pro
265 270 275
ctg ctg ctg gat ggc aga gag ttt tca gtt ccg atg gcg acg acg gag 916
Leu Leu Leu Asp Gly Arg Glu Phe Ser Val Pro Met Ala Thr Thr Glu
280 285 290 295
ggc tgt ttg gtg gcc agc aca aac agg gga tgc aag gca att ttc gcc 964
Gly Cys Leu Val Ala Ser Thr Asn Arg Gly Cys Lys Ala Ile Phe Ala
300 305 310
tca ggt ggt gca tcc agc gta gtt tat agg gac ggg atg acc agg gcg 1012
Ser Gly Gly Ala Ser Ser Val Val Tyr Arg Asp Gly Met Thr Arg Ala
315 320 325
ccg gct gtc agg ttc ggc tca gcc aag agg gct gca gaa ttg aag ctg 1060
Pro Ala Val Arg Phe Gly Ser Ala Lys Arg Ala Ala Glu Leu Lys Leu
330 335 340
ttt ttg gag aat ccc gtc aat ttc gaa acc tta gct ctt gtt ttc aac 1108
Phe Leu Glu Asn Pro Val Asn Phe Glu Thr Leu Ala Leu Val Phe Asn
345 350 355
aaa tcc agt cga ttc gcg aag ctg cag agc atc aaa tgc gcc att gct 1156
Lys Ser Ser Arg Phe Ala Lys Leu Gln Ser Ile Lys Cys Ala Ile Ala
360 365 370 375
gga aag aat ctg tac gcg agg ttc tcc tgc agc act ggg gat gcc atg 1204
Gly Lys Asn Leu Tyr Ala Arg Phe Ser Cys Ser Thr Gly Asp Ala Met
380 385 390
gga atg aat atg gtc tcc aaa ggt gtg cag aac gtt ttg gat ttc ctt 1252
Gly Met Asn Met Val Ser Lys Gly Val Gln Asn Val Leu Asp Phe Leu
395 400 405
cag gcg gac ttc ccc gac atg gat gtg att ggc atc tct gga aac tac 1300
Gln Ala Asp Phe Pro Asp Met Asp Val Ile Gly Ile Ser Gly Asn Tyr
410 415 420
tgt tca gac aag aag ccg gct gca gta aac tgg att gaa ggt cgt gga 1348
Cys Ser Asp Lys Lys Pro Ala Ala Val Asn Trp Ile Glu Gly Arg Gly
425 430 435
aag tct gtg gtt tgt gag gct atc ata aag gag gag gtg gta aag aag 1396
Lys Ser Val Val Cys Glu Ala Ile Ile Lys Glu Glu Val Val Lys Lys
440 445 450 455
gtg ctg aag acg gag gtt gct tcc ctg gtt gag ctt aac atg ctg aag 1444
Val Leu Lys Thr Glu Val Ala Ser Leu Val Glu Leu Asn Met Leu Lys
460 465 470
aac ctg gtg ggg tct gcc atg gct ggt gct ctg gga ggc ttc aac gcc 1492
Asn Leu Val Gly Ser Ala Met Ala Gly Ala Leu Gly Gly Phe Asn Ala
475 480 485
cat gcc agt aac atc gta tca gcc att tat ata gca act gga caa gac 1540
His Ala Ser Asn Ile Val Ser Ala Ile Tyr Ile Ala Thr Gly Gln Asp
490 495 500
cca gcg cag aac att gag agc tcc cac tgc atc acc atg atg gaa gct 1588
Pro Ala Gln Asn Ile Glu Ser Ser His Cys Ile Thr Met Met Glu Ala
505 510 515
gtc aac ggt gga aag gac ctt cac gtc tca gtg acg atg cca tct gta 1636
Val Asn Gly Gly Lys Asp Leu His Val Ser Val Thr Met Pro Ser Val
520 525 530 535
gag gta ggt aca gtt gga ggt gga acc caa ctt gca tct cag tcg gca 1684
Glu Val Gly Thr Val Gly Gly Gly Thr Gln Leu Ala Ser Gln Ser Ala
540 545 550
tgt ctg aac ctt ctt ggg gtg aag ggt gcc agc aaa gag tta ccg gga 1732
Cys Leu Asn Leu Leu Gly Val Lys Gly Ala Ser Lys Glu Leu Pro Gly
555 560 565
tcc aat gca agg ctg ttg gct acc gtt gta gct ggt tct gta ctg gca 1780
Ser Asn Ala Arg Leu Leu Ala Thr Val Val Ala Gly Ser Val Leu Ala
570 575 580
ggg gag ctc tcc ctt atg tct gca ctt gcc gcg gga cag ctt gtc aag 1828
Gly Glu Leu Ser Leu Met Ser Ala Leu Ala Ala Gly Gln Leu Val Lys
585 590 595
agt cac atg aag tac aat agg tcc tct aac aag acc act cat ggt caa 1876
Ser His Met Lys Tyr Asn Arg Ser Ser Asn Lys Thr Thr His Gly Gln
600 605 610 615
gtt acc gaa act cca ttc aac cca act ctt gct tct tct tat taa 1921
Val Thr Glu Thr Pro Phe Asn Pro Thr Leu Ala Ser Ser Tyr
620 625
attggtgtct ttgaggaata aggagagaca tggaagctaa cccaaactaa agaaacactc 1981
attacatgtg tatttgacta ctatttgtcg ttatcaacaa cttttactat accatttgga 2041
cgaaaaaaaa aaaaaa 2057
<210>12
<211>629
<212>PRT
<213>杜仲
<400>12
Met Trp Asn Ser Asn Lys Asn His Ile Arg His Ser Leu Ser Pro Ser
1 5 10 15
Arg Phe Ser Arg Tyr Thr Tyr Ile Asp Ser Ser Thr Glu Ser Lys Ser
20 25 30
Met Asp Val Arg Arg Arg Ser Met Lys Pro Thr Ala Val His His Thr
35 40 45
Pro Thr His Ser Tyr Ala Leu Lys Pro Pro Gln Lys Asp Asp Asp Asp
50 55 60
Leu Pro Lys Ala Ser Asp Ala Leu Pro Phe His Leu Tyr Leu Thr Asn
65 70 75 80
Ala Val Phe Phe Thr Leu Phe Phe Ser Val Val Tyr Tyr Leu Leu Ser
85 90 95
Arg Trp Arg Glu Lys Ile Arg Asn Ser Thr Pro Leu His Ile Val Ser
100 105 110
Leu Ser Glu Ile Ala Ala Ile Val Ala Phe Val Ala Ser Phe Ile Tyr
115 120 125
Leu Leu Gly Phe Phe Gly Ile Gly Phe Val Gln Ser Phe Ala Pro Arg
130 135 140
Ser Ser His Asp His Asp Val Asp Asp Glu Ile Trp Val Asn Asp Asp
145 150 155 160
Ile Ile Arg Lys Asp Asp Asp Gly Arg Thr Gly Pro Cys Ala Val Asn
165 170 175
Cys Pro Leu Pro Lys Pro Tyr Asn Lys Val Val Val Glu Glu Ser Pro
180 185 190
Asp Pro Pro Leu Val Ser Ser Ala Glu Asp Glu Glu Ile Ile Asn Ser
195 200 205
Val Val Glu Gly Lys Ile Pro Ser Tyr Ala Leu Glu Ser Lys Leu Ser
210 215 220
Asp Cys Arg Arg Ala Ala Ala Val Arg Arg Glu Ala Leu Gln Arg Met
225 230 235 240
Thr Gly Lys Ser Leu Val Gly Leu Pro Leu Glu Gly Phe Asp Tyr Glu
245 250 255
Ser Ile Leu Gly Gln Cys Cys Glu Met Pro Val Gly Tyr Val Gln Ile
260 265 270
Pro Val Gly Ile Ala Gly Pro Leu Leu Leu Asp Gly Arg Glu Phe Ser
275 280 285
Val Pro Met Ala Thr Thr Glu Gly Cys Leu Val Ala Ser Thr Asn Arg
290 295 300
Gly Cys Lys Ala Ile Phe Ala Ser Gly Gly Ala Ser Ser Val Val Tyr
305 310 315 320
Arg Asp Gly Met Thr Arg Ala Pro Ala Val Arg Phe Gly Ser Ala Lys
325 330 335
Arg Ala Ala Glu Leu Lys Leu Phe Leu Glu Asn Pro Val Asn Phe Glu
340 345 350
Thr Leu Ala Leu Val Phe Asn Lys Ser Ser Arg Phe Ala Lys Leu Gln
355 360 365
Ser Ile Lys Cys Ala Ile Ala Gly Lys Asn Leu Tyr Ala Arg Phe Ser
370 375 380
Cys Ser Thr Gly Asp Ala Met Gly Met Asn Met Val Ser Lys Gly Val
385 390 395 400
Gln Asn Val Leu Asp Phe Leu Gln Ala Asp Phe Pro Asp Met Asp Val
405 410 415
Ile Gly Ile Ser Gly Asn Tyr Cys Ser Asp Lys Lys Pro Ala Ala Val
420 425 430
Asn Trp Ile Glu Gly Arg Gly Lys Ser Val Val Cys Glu Ala Ile Ile
435 440 445
Lys Glu Glu Val Val Lys Lys Val Leu Lys Thr Glu Val Ala Ser Leu
450 455 460
Val Glu Leu Asn Met Leu Lys Asn Leu Val Gly Ser Ala Met Ala Gly
465 470 475 480
Ala Leu Gly Gly Phe Asn Ala His Ala Ser Asn Ile Val Ser Ala Ile
485 490 495
Tyr Ile Ala Thr Gly Gln Asp Pro Ala Gln Asn Ile Glu Ser Ser His
500 505 510
Cys Ile Thr Met Met Glu Ala Val Asn Gly Gly Lys Asp Leu His Val
515 520 525
Ser Val Thr Met Pro Ser Val Glu Val Gly Thr Val Gly Gly Gly Thr
530 535 540
Gln Leu Ala Ser Gln Ser Ala Cys Leu Asn Leu Leu Gly Val Lys Gly
545 550 555 560
Ala Ser Lys Glu Leu Pro Gly Ser Asn Ala Arg Leu Leu Ala Thr Val
565 570 575
Val Ala Gly Ser Val Leu Ala Gly Glu Leu Ser Leu Met Ser Ala Leu
580 585 590
Ala Ala Gly Gln Leu Val Lys Ser His Met Lys Tyr Asn Arg Ser Ser
595 600 605
Asn Lys Thr Thr His Gly Gln Val Thr Glu Thr Pro Phe Asn Pro Thr
610 61 5 620
Leu Ala Ser Ser Tyr
625
<210>13
<211>2225
<212>DNA
<213>杜仲
<220>
<221>CDS
<222>(60)..(1835)
<223>HMG-CoA还原酶
<400>13
aagaagagag caccatttct ctctcttctc ccaacaaaca cacactagag agagagaga 59
atg gac aat cgc cgg cgt caa tcc aag cca gcc cgg ccg gca aag atc 107
Met Asp Asn Arg Arg Arg Gln Ser Lys Pro Ala Arg Pro Ala Lys Ile
1 5 10 15
gcc gcc ggc gac tct ctc aag cca cat aaa ctc caa cac tac caa act 155
Ala Ala Gly Asp Ser Leu Lys Pro His Lys Leu Gln His Tyr Gln Thr
20 25 30
tcc act gtc aaa gcc tcc gac gcc ctt cct ctt ccc ctg tac ctc atg 203
Ser Thr Val Lys Ala Ser Asp Ala Leu Pro Leu Pro Leu Tyr Leu Met
35 40 45
aac ggc ttg ttc ttc acc atg ttc ttc tcc gtc atg tac ttc ctc ctc 251
Asn Gly Leu Phe Phe Thr Met Phe Phe Ser Val Met Tyr Phe Leu Leu
50 55 60
cac cgc tgg cgt gaa aag atc cgc aat tcc act ccc ctc cac gtc gtc 299
His Arg Trp Arg Glu Lys Ile Arg Asn Ser Thr Pro Leu His Val Val
65 70 75 80
acg ctt tcc gag ctg gcg gcc gtg gtt tcc tta atc gcc tcc gcg att 347
Thr Leu Ser Glu Leu Ala Ala Val Val Ser Leu Ile Ala Ser Ala Ile
85 90 95
tac ctc ctg ggc ttt ttc ggc atc gat ttc gtg cag tcc ttg att ttc 395
Tyr Leu Leu Gly Phe Phe Gly Ile Asp Phe Val Gln Ser Leu Ile Phe
100 105 110
cgg agc tcc cct gaa gga tgg gat aca gag gac gag acg tcg gag caa 443
Arg Ser Ser Pro Glu Gly Trp Asp Thr Glu Asp Glu Thr Ser Glu Gln
115 120 125
caa ttg atc gtg gaa gac agc cgc ggc ggt cca gcg acg tcg att ggc 491
Gln Leu Ile Val Glu Asp Ser Arg Gly Gly Pro Ala Thr Ser Ile Gly
130 135 140
tgt tcg ctt cct ccg att gca ccc aag gcg ttg gtt ccg tcc atg gag 539
Cys Ser Leu Pro Pro Ile Ala Pro Lys Ala Leu Val Pro Ser Met Glu
145 150 155 160
aca gcg gaa ttc gcc gtc cct caa ccg ctg acg gag gaa gac gag gag 587
Thr Ala Glu Phe Ala Val Pro Gln Pro Leu Thr Glu Glu Asp Glu Glu
165 170 175
ata atc aaa tca gtg gtg gcc ggg aca act ccg tcg tac tcc ttg gag 635
Ile Ile Lys Ser Val Val Ala Gly Thr Thr Pro Ser Tyr Ser Leu Glu
180 185 190
tcg aag ctc ggg gac tgc cgg aga gca gcc gca att cga agg gag tca 683
Ser Lys Leu Gly Asp Cys Arg Arg Ala Ala Ala Ile Arg Arg Glu Ser
195 200 205
ctc cag aga atc acc gga aag tct ctg gcg ggg ctt ccg tta gac gga 731
Leu Gln Arg Ile Thr Gly Lys Ser Leu Ala Gly Leu Pro Leu Asp Gly
210 215 220
ttc gat tac aat tcg att ctg gga cag tgc tgt gag atg ccg gtg ggg 779
Phe Asp Tyr Asn Ser Ile Leu Gly Gln Cys Cys Glu Met Pro Val Gly
225 230 235 240
tac att cag att ccg gtg gga atc gcc ggc cca ttg ttg ctg gac gga 827
Tyr Ile Gln Ile Pro Val Gly Ile Ala Gly Pro Leu Leu Leu Asp Gly
245 250 255
gct gaa tat tct gtg ccg atg gcg aca acg gag ggt tgc ctg gtg gcc 875
Ala Glu Tyr Ser Val Pro Met Ala Thr Thr Glu Gly Cys Leu Val Ala
260 265 270
agc acc aac agg gga tgc aag gcc atc tat ctc tcc ggc gga gcc acc 923
Ser Thr Asn Arg Gly Cys Lys Ala Ile Tyr Leu Ser Gly Gly Ala Thr
275 280 285
tct atg ttg ttg gcg gac gcc atg acc aga gct ccg gtg gtt aga ttc 971
Ser Met Leu Leu Ala Asp Ala Met Thr Arg Ala Pro Val Val Arg Phe
290 295 300
gga acc gcc aaa aga gcc gcc gag ttg aaa ttc ttc ctt gaa gat ccc 1019
Gly Thr Ala Lys Arg Ala Ala Glu Leu Lys Phe Phe Leu Glu Asp Pro
305 310 315 320
atg aat ttc gac act ctc tct gtt gtc ttc aac aaa tcc agc aga ttt 1067
Met Asn Phe Asp Thr Leu Ser Val Val Phe Asn Lys Ser Ser Arg Phe
325 330 335
ggt agg cta cag ggc att cag tgt gca atc gcc gga aaa aat ctg tac 1115
Gly Arg Leu Gln Gly Ile Gln Cys Ala Ile Ala Gly Lys Asn Leu Tyr
340 345 350
atc cgg ttc agc tgc agc acc ggc gac gcc atg ggg atg aac atg gtc 1163
Ile Arg Phe Ser Cys Ser Thr Gly Asp Ala Met Gly Met Asn Met Val
355 360 365
tcc aaa ggc gtt cag aac gtt cta gat ttc ctt cag gca gat ttc ccc 1211
Ser Lys Gly Val Gln Asn Val Leu Asp Phe Leu Gln Ala Asp Phe Pro
370 375 380
gac atg gac gtc atc ggt atc tcc ggc aat tac tgc tcc gac aag aaa 1259
Asp Met Asp Val Ile Gly Ile Ser Gly Asn Tyr Cys Ser Asp Lys Lys
385 390 395 400
cca gcg gcg gtg aac tgg atc cag ggc cgc ggt aag tcg gtg gtc tgc 1307
Pro Ala Ala Val Asn Trp Ile Gln Gly Arg Gly Lys Ser Val Val Cys
405 410 415
gaa gcc gtc atc aag gaa gag gtg gtg aag aag gtg ctg aag acc aac 1355
Glu Ala Val Ile Lys Glu Glu Val Val Lys Lys Val Leu Lys Thr Asn
420 425 430
gtc gcg gcg ctg gtg gaa ctc aac atg ctg aaa aac ctc gcc gga tcg 1403
Val Ala Ala Leu Val Glu Leu Asn Met Leu Lys Asn Leu Ala Gly Ser
435 440 445
gcg gtt gcc ggt tca ctc ggt ggg ttc aac gcc cac gcc agt aat att 1451
Ala Val Ala Gly Ser Leu Gly Gly Phe Asn Ala His Ala Ser Asn Ile
450 455 460
gtc tcc gcc gtg tat att gcc acc ggc caa gat ccg gca cag aac gtg 1499
Val Ser Ala Val Tyr Ile Ala Thr Gly Gln Asp Pro Ala Gln Asn Val
465 470 475 480
gag agc tcc cac tgc atc acc atg atg gag cct ctc aac gac ggg aag 1547
Glu Ser Ser His Cys Ile Thr Met Met Glu Pro Leu Asn Asp Gly Lys
485 490 495
gac ctt cat gtc tcc gtc acc atg cct tcc att gag gtg gga aca gta 1595
Asp Leu His Val Ser Val Thr Met Pro Ser Ile Glu Val Gly Thr Val
500 505 510
gga gga gga aca cag ttg gcg tca cag tcg gca tgc ttg aac cta ctg 1643
Gly Gly Gly Thr Gln Leu Ala Ser Gln Ser Ala Cys Leu Asn Leu Leu
515 520 525
gga gta aag ggt gcg aac aag gag tcg cca gga tca aac gca agg cta 1691
Gly Val Lys Gly Ala Asn Lys Glu Ser Pro Gly Ser Asn Ala Arg Leu
530 535 540
ttg gcc acc atc gtc gcc ggt tct gtt ctc gcc ggc gaa tta tcc cta 1739
Leu Ala Thr Ile Val Ala Gly Ser Val Leu Ala Gly Glu Leu Ser Leu
545 550 555 560
atg tcg gca atc gcc gcc ggt caa ctg gtg aaa agc cac atg aag tac 1787
Met Ser Ala Ile Ala Ala Gly Gln Leu Val Lys Ser His Met Lys Tyr
565 570 575
aac aga tcc acc aaa gac atc aca aag gtc gtc tcc acc acc aac taa 1835
Asn Arg Ser Thr Lys Asp Ile Thr Lys Val Val Ser Thr Thr Asn
580 585 590
aacataaaaa aggcaactac tttttattta tttgtataaa gaaaaaatag acccccccaa 1895
aagggaaaaa ataatgatca agaaaaagct cgaaataaaa atctccatta ctgaaagtgg 1955
tagagagaga aagagggagg agaggatttg tggtgtattt tctagtacag aggggaagaa 2015
agtagagcgt accatgatga agacgaggtg gtggtggtcg tcgtttaatt ttttttttta 2075
aattctgaaa aatcataaat gtcagcagag ctttgttgac atgaagatgc tcttctcgca 2135
cgatgcatgt ggttcttgat gtctttgtaa attaattatc aaagggaaaa aggcactgcc 2195
atttccttta ccatcgaaaa aaaaaaaaaa 2225
<210>14
<211>591
<212>PRT
<213>杜仲
<400>14
Met Asp Asn Arg Arg Arg Gln Ser Lys Pro Ala Arg Pro Ala Lys Ile
1 5 10 15
Ala Ala Gly Asp Ser Leu Lys Pro His Lys Leu Gln His Tyr Gln Thr
20 25 30
Ser Thr Val Lys Ala Ser Asp Ala Leu Pro Leu Pro Leu Tyr Leu Met
35 40 45
Asn Gly Leu Phe Phe Thr Met Phe Phe Ser Val Met Tyr Phe Leu Leu
50 55 60
His Arg Trp Arg Glu Lys Ile Arg Asn Ser Thr Pro Leu His Val Val
65 70 75 80
Thr Leu Ser Glu Leu Ala Ala Val Val Ser Leu Ile Ala Ser Ala Ile
85 90 95
Tyr Leu Leu Gly Phe Phe Gly Ile Asp Phe Val Gln Ser Leu Ile Phe
100 105 110
Arg Ser Ser Pro Glu Gly Trp Asp Thr Glu Asp Glu Thr Ser Glu Gln
115 120 125
Gln Leu Ile Val Glu Asp Ser Arg Gly Gly Pro Ala Thr Ser Ile Gly
130 135 140
Cys Ser Leu Pro Pro Ile Ala Pro Lys Ala Leu Val Pro Ser Met Glu
145 150 155 160
Thr Ala Glu Phe Ala Val Pro Gln Pro Leu Thr Glu Glu Asp Glu Glu
165 170 175
Ile Ile Lys Ser Val Val Ala Gly Thr Thr Pro Ser Tyr Ser Leu Glu
180 185 190
Ser Lys Leu Gly Asp Cys Arg Arg Ala Ala Ala Ile Arg Arg Glu Ser
195 200 205
Leu Gln Arg Ile Thr Gly Lys Ser Leu Ala Gly Leu Pro Leu Asp Gly
210 215 220
Phe Asp Tyr Asn Ser Ile Leu Gly Gln Cys Cys Glu Met Pro Val Gly
225 230 235 240
Tyr Ile Gln Ile Pro Val Gly Ile Ala Gly Pro Leu Leu Leu Asp Gly
245 250 255
Ala Glu Tyr Ser Val Pro Met Ala Thr Thr Glu Gly Cys Leu Val Ala
260 265 270
Ser Thr Asn Arg Gly Cys Lys Ala Ile Tyr Leu Ser Gly Gly Ala Thr
275 280 285
Ser Met Leu Leu Ala Asp Ala Met Thr Arg Ala Pro Val Val Arg Phe
290 295 300
Gly Thr Ala Lys Arg Ala Ala Glu Leu Lys Phe Phe Leu Glu Asp Pro
305 310 315 320
Met Asn Phe Asp Thr Leu Ser Val Val Phe Asn Lys Ser Ser Arg Phe
325 330 335
Gly Arg Leu Gln Gly Ile Gln Cys Ala Ile Ala Gly Lys Asn Leu Tyr
340 345 350
Ile Arg Phe Ser Cys Ser Thr Gly Asp Ala Met Gly Met Asn Met Val
355 360 365
Ser Lys Gly Val Gln Asn Val Leu Asp Phe Leu Gln Ala Asp Phe Pro
370 375 380
Asp Met Asp Val Ile Gly Ile Ser Gly Asn Tyr Cys Ser Asp Lys Lys
385 390 395 400
Pro Ala Ala Val Asn Trp Ile Gln Gly Arg Gly Lys Ser Val Val Cys
405 410 415
Glu Ala Val Ile Lys Glu Glu Val Val Lys Lys Val Leu Lys Thr Asn
420 425 430
Val Ala Ala Leu Val Glu Leu Asn Met Leu Lys Asn Leu Ala Gly Ser
435 440 445
Ala Val Ala Gly Ser Leu Gly Gly Phe Asn Ala His Ala Ser Asn Ile
450 455 460
Val Ser Ala Val Tyr Ile Ala Thr Gly Gln Asp Pro Ala Gln Asn Val
465 470 475 480
Glu Ser Ser His Cys Ile Thr Met Met Glu Pro Leu Asn Asp Gly Lys
485 490 495
Asp Leu His Val Ser Val Thr Met Pro Ser Ile Glu Val Gly Thr Val
500 505 510
Gly Gly Gly Thr Gln Leu Ala Ser Gln Ser Ala Cys Leu Asn Leu Leu
515 520 525
Gly Val Lys Gly Ala Asn Lys Glu Ser Pro Gly Ser Asn Ala Arg Leu
530 535 540
Leu Ala Thr Ile Val Ala Gly Ser Val Leu Ala Gly Glu Leu Ser Leu
545 550 555 560
Met Ser Ala Ile Ala Ala Gly Gln Leu Val Lys Ser His Met Lys Tyr
565 570 575
Asn Arg Ser Thr Lys Asp Ile Thr Lys Val Val Ser Thr Thr Asn
580 585 590
<210>15
<211>1341
<212>DNA
<213>杜仲
<220>
<221>CDS
<222>(61)..(1224)
<223>甲羟戊酸激酶
<400>15
ctcgttcgta ctgaaatcaa accgagaaag caactagtca aataattggg gcggagacga 60
atg gag gtg aga gct aga gca cct ggg aaa att att atc gcc ggc gag 108
Met Glu Val Arg Ala Arg Ala Pro Gly Lys Ile Ile Ile Ala Gly Glu
1 5 10 15
cac gcc gtc gtc cac gga tca acg gcc gtt gcc gcc tca gtc gat ctc 156
His Ala Val Val His Gly Ser Thr Ala Val Ala Ala Ser Val Asp Leu
20 25 30
tac acc tgc gtc tcc ctt cgc ttc ccc acc cct tct gac aat gat gac 204
Tyr Thr Cys Val Ser Leu Arg Phe Pro Thr Pro Ser Asp Asn Asp Asp
35 40 45
tcc ttg aag ttc cac ctc aaa gat ttg gat tta gag ttt tcc tgg cca 252
Ser Leu Lys Phe His Leu Lys Asp Leu Asp Leu Glu Phe Ser Trp Pro
50 55 60
att gaa aga gtg aaa gaa gca ctt tct gaa tta gga agt tgc gcc cct 300
Ile Glu Arg Val Lys Glu Ala Leu Ser Glu Leu Gly Ser Cys Ala Pro
65 70 75 80
tca tca cct acc tca tgt tca ctg ggg acc atg aaa gca att gca gca 348
Ser Ser Pro Thr Ser Cys Ser Leu Gly Thr Met Lys Ala Ile Ala Ala
85 90 95
cta gtt gaa gaa cat act att cca gag gca aac att gga ctt gct gct 396
Leu Val Glu Glu His Thr Ile Pro Glu Ala Asn Ile Gly Leu Ala Ala
100 105 110
ggt gtt tca gct ttt tta tgg ctt tat atc tct atc cag ggt tat aaa 444
Gly Val Ser Ala Phe Leu Trp Leu Tyr Ile Ser Ile Gln Gly Tyr Lys
115 120 125
cct gga aat gtg gtt atc act tcc gag ctt cca ctt ggc tca ggc ttg 492
Pro Gly Asn Val Val Ile Thr Ser Glu Leu Pro Leu Gly Ser Gly Leu
130 135 140
ggt tca tct gcc tct ttc tgt gtg gca ctt gca gga gcc ctg ctt gct 540
Gly Ser Ser Ala Ser Phe Cys Val Ala Leu Ala Gly Ala Leu Leu Ala
145 150 155 160
ttg tca gat tct gtg tct ctg gat ttg agc cat gaa ggg tgg ttg atg 588
Leu Ser Asp Ser Val Ser Leu Asp Leu Ser His Glu Gly Trp Leu Met
165 170 175
ttt gga gaa agt gag ttg gaa ctg gtt aac aaa tgg gct ttc gaa gga 636
Phe Gly Glu Ser Glu Leu Glu Leu Val Asn Lys Trp Ala Phe Glu Gly
180 185 190
gaa aag ata ctc cat ggg aag cca tct ggg ata gac aac aca gtc agc 684
Glu Lys Ile Leu His Gly Lys Pro Ser Gly Ile Asp Asn Thr Val Ser
195 200 205
aca tat ggt aac atg atc aag ttc aag tta ggt gat ttg acg cgc atc 732
Thr Tyr Gly Asn Met Ile Lys Phe Lys Leu Gly Asp Leu Thr Arg Ile
210 215 220
aat gcg ggc atg cca ctt aaa atg ctt ata act aat act aaa gtt gga 780
Asn Ala Gly Met Pro Leu Lys Met Leu Ile Thr Asn Thr Lys Val Gly
225 230 235 240
cgg aac aca aaa gcg ttg gtt gct agc gta tca gag agg aca gta agg 828
Arg Asn Thr Lys Ala Leu Val Ala Ser Val Ser Glu Arg Thr Val Arg
245 250 255
cat cct aat gct atg gcc tct gtg ttt agt gct gtt gat tca atc agc 876
His Pro Asn Ala Met Ala Ser Val Phe Ser Ala Val Asp Ser Ile Ser
260 265 270
aat gaa ttg gca aca ata atc caa tct cct gct tct gat gat ttt gcc 924
Asn Glu Leu Ala Thr Ile Ile Gln Ser Pro Ala Ser Asp Asp Phe Ala
275 280 285
ata acc gag aag gaa gag aaa ctt gaa gaa cta atg gaa atg aat caa 972
Ile Thr Glu Lys Glu Glu Lys Leu Glu Glu Leu Met Glu Met Asn Gln
290 295 300
ggg ctg ctg caa tgc atg gga gtt agc cat tct tct ata gaa act gtg 1020
Gly Leu Leu Gln Cys Met Gly Val Ser His Ser Ser Ile Glu Thr Val
305 310 315 320
ctt cgg aca aca ttg aaa tac aaa tta gct tcg aag tta act gga gct 1068
Leu Arg Thr Thr Leu Lys Tyr Lys Leu Ala Ser Lys Leu Thr Gly Ala
325 330 335
ggg ggt gga ggg tgc gtg ctg aca ctg ctt cca acc tta ctg tcg gaa 1116
Gly Gly Gly Gly Cys Val Leu Thr Leu Leu Pro Thr Leu Leu Ser Glu
340 345 350
acg att gtt ggc aga gta att agt gag cta gag gcc tgc ggt ttc caa 1164
Thr Ile Val Gly Arg Val Ile Ser Glu Leu Glu Ala Cys Gly Phe Gln
355 360 365
tgt tta ctt gcc gga atc ggc ggg aaa ggt gtt cag atc tgc ttt ggc 1212
Cys Leu Leu Ala Gly Ile Gly Gly Lys Gly Val Gln Ile Cys Phe Gly
370 375 380
ggc tcg tct tga tttcgctccg gagaatcata gcaaaataag tctcagttac 1264
Gly Ser Ser
385
aaaaactcca aatattgatg agttgaataa tatatatata tattgtttgc tttggcaatc 1324
ctaaaaaaaa aaaaaaa 1341
<210>16
<211>387
<212>PRT
<213>杜仲
<400>16
Met Glu Val Arg Ala Arg Ala Pro Gly Lys Ile Ile Ile Ala Gly Glu
1 5 10 15
His Ala Val Val His Gly Ser Thr Ala Val Ala Ala Ser Val Asp Leu
20 25 30
Tyr Thr Cys Val Ser Leu Arg Phe Pro Thr Pro Ser Asp Asn Asp Asp
35 40 45
Ser Leu Lys Phe His Leu Lys Asp Leu Asp Leu Glu Phe Ser Trp Pro
50 55 60
Ile Glu Arg Val Lys Glu Ala Leu Ser Glu Leu Gly Ser Cys Ala Pro
65 70 75 80
Ser Ser Pro Thr Ser Cys Ser Leu Gly Thr Met Lys Ala Ile Ala Ala
85 90 95
Leu Val Glu Glu His ThrIle Pro Glu Ala Asn Ile Gly Leu Ala Ala
100 105 110
Gly Val Ser Ala Phe Leu Trp Leu Tyr Ile Ser Ile Gln Gly Tyr Lys
115 120 125
Pro Gly Asn Val Val Ile Thr Ser Glu Leu Pro Leu Gly Ser Gly Leu
130 135 140
Gly Ser Ser Ala Ser Phe Cys Val Ala Leu Ala Gly Ala Leu Leu Ala
145 150 155 160
Leu Ser Asp Ser Val Ser Leu Asp Leu Ser His Glu Gly Trp Leu Met
165 170 175
Phe Gly Glu Ser Glu Leu Glu Leu Val Asn Lys Trp Ala Phe Glu Gly
180 185 190
Glu Lys Ile Leu His Gly Lys Pro Ser Gly Ile Asp Asn Thr Val Ser
195 200 205
Thr Tyr Gly Asn Met Ile Lys Phe Lys Leu Gly Asp Leu Thr Arg Ile
210 215 220
Asn Ala Gly Met Pro Leu Lys Met Leu Ile Thr Asn Thr Lys Val Gly
225 230 235 240
Arg Asn Thr Lys Ala Leu Val Ala Ser Val Ser Glu Arg Thr Val Arg
245 250 255
His Pro Asn Ala Met Ala Ser Val Phe Ser Ala Val Asp Ser Ile Ser
260 265 270
Asn Glu Leu Ala Thr Ile Ile Gln Ser Pro Ala Ser Asp Asp Phe Ala
275 280 285
Ile Thr Glu Lys Glu Glu Lys Leu Glu Glu Leu Met Glu Met Asn Gln
290 295 300
Gly Leu Leu Gln Cys Met Gly Val Ser His Ser Ser Ile Glu Thr Val
305 310 315 320
Leu Arg Thr Thr Leu Lys Tyr Lys Leu Ala Ser Lys Leu Thr Gly Ala
325 330 335
Gly Gly Gly Gly Cys Val Leu Thr Leu Leu Pro Thr Leu Leu Ser Glu
340 345 350
Thr Ile Val Gly Arg Val Ile Ser Glu Leu Glu Ala Cys Gly Phe Gln
355 360 365
Cys Leu Leu Ala Gly Ile Gly Gly Lys Gly Val Gln Ile Cys Phe Gly
370 375 380
Gly Ser Ser
385
<210>17
<211>2432
<212>DNA
<213>杜仲
<220>
<221>CDS
<222>(682)..(2202)
<223>磷酸甲羟戊酸激酶
<400>17
tatagttgaa cttctatatt atgatatcat gtcgatttta tagtacattt ttgcctggaa 60
atattacacc ggacgacatt ttagtgaaat ttcactaacc aacattataa gtattgagct 120
catcatactc catgcttaaa caagatttca gtgaaatttc actaaatact tatacaagaa 180
ccagcattat aaacacaaac ctcttcaaat cttttatgat caagaaataa cacaagcgtt 240
tggattgtcc tcagcactgt gaacatgata gtattgtgtc atgtatgggt tataagcctt 300
ataggccttc acaagttctt ctatttgttc gttctggtct tttttcttgt catcgtcttt 360
ctttccgtct tctttctttc cctcatcctt ctttccatca tctttctttt cctcctttgc 420
aggtccaact gtcaatatct ctgcatgata taagttttct cactttactc actacttcga 480
cgggatcgac gaatttcata tttgaacgcc gaagcttagt ttgtgtcgat cgactgcgaa 540
tcggaggagg acgattgtcg atcgccggca caactacatt ctccggcgta atatccattt 600
tccgtgaatt aaccggattc ggttgtattc cagatcaatt gaatctcagc tttccaagat 660
cagattgaaa tttctagaga a atg gct gta gtt gca tct gct ccc ggg aag 711
Met Ala Val Val Ala Ser Ala Pro Gly Lys
1 5 10
gtt ttg atg act ggg ggt tac ctt att ctt gag agg cct aat gct ggg 759
Val Leu Met Thr Gly Gly Tyr Leu Ile Leu Glu Arg Pro Asn Ala Gly
15 20 25
att gtg ctc agt aca aat gct cga ttt tat gca att gtg aaa cca ctt 807
Ile Val Leu Ser Thr Asn Ala Arg Phe Tyr Ala Ile Val Lys Pro Leu
30 35 40
tat gaa gaa gtt aga cct gac agt tgg gct tgg gca tgg aca gac gta 855
Tyr Glu Glu Val Arg Pro Asp Ser Trp Ala Trp Ala Trp Thr Asp Val
45 50 55
aaa tta aca tct cca caa atg gca aga gaa act atg tat aag ttg tcg 903
Lys Leu Thr Ser Pro Gln Met Ala Arg Glu Thr Met Tyr Lys Leu Ser
60 65 70
ctt aaa cat att ata acc cag tgt gtc tct cca agt cag tca ggg aat 951
Leu Lys His Ile Ile Thr Gln Cys Val Ser Pro Ser Gln Ser Gly Asn
75 80 85 90
cca ttc gta gaa cat gcg gtg caa tat gct gta gca gca gca cat gca 999
Pro Phe Val Glu His Ala Val Gln Tyr Ala Val Ala Ala Ala His Ala
95 100 105
att ttt gac aat aac aaa aaa gat gca ttg cag aaa cgg ctt ttg caa 1047
Ile Phe Asp Asn Asn Lys Lys Asp Ala Leu Gln Lys Arg Leu Leu Gln
110 115 120
ggc ctt gac att aca ata tta ggt tgt aat gac ttc tat tca tac cgg 1095
Gly Leu Asp Ile Thr Ile Leu Gly Cys Asn Asp Phe Tyr Ser Tyr Arg
125 130 135
ggc gag att gaa gca cgt gga ctt cct ttg aca cct gat tct ttg gct 1143
Gly Glu Ile Glu Ala Arg Gly Leu Pro Leu Thr Pro Asp Ser Leu Ala
140 145 150
tct ctt cct cca ttt tct tct att acc ttt aat gct gaa gag tct agt 1191
Ser Leu Pro Pro Phe Ser Ser Ile Thr Phe Asn Ala Glu Glu Ser Ser
155 160 165 170
gaa aag aat tgc aag cct gaa gtt gcc aaa act ggg ttg gga tct tct 1239
Glu Lys Asn Cys Lys Pro Glu Val Ala Lys Thr Gly Leu Gly Ser Ser
175 180 185
gca gct atg aca acc gca gtg gtt gct gct tta ctt cat tac ctt gaa 1287
Ala Ala Met Thr Thr Ala Val Val Ala Ala Leu Leu His Tyr Leu Glu
190 195 200
gtt gtg gac ctt tcc tcc ttg act gaa gct caa aat cag aat gtt gag 1335
Val Val Asp Leu Ser Ser Leu Thr Glu Ala Gln Asn Gln Asn Val Glu
205 210 215
caa gat ctt gat ttg gta cat gtg ata gct caa act gca cac tgt att 1383
Gln Asp Leu Asp Leu Val His Val Ile Ala Gln Thr Ala His Cys Ile
220 225 230
gcc cag gga aaa gtt gga agt ggg ttt gat gtc agt tct gca gtt tat 1431
Ala Gln Gly Lys Val Gly Ser Gly Phe Asp Val Ser Ser Ala Val Tyr
235 240 245 250
ggc agt caa cat tac gtc cga ttt tca cct gac gta ctt tct tct gct 1479
Gly Ser Gln His Tyr Val Arg Phe Ser Pro Asp Val Leu Ser Ser Ala
255 260 265
cag ggt gct gta aaa ggc atg ccg cta gaa gaa gtc att tca gat gtc 1527
Gln Gly Ala Val Lys Gly Met Pro Leu Glu Glu Val Ile Ser Asp Val
270 275 280
ctg aaa gcc aag tgg gac cat gag agg act aaa ttc tca ttg cct cca 1575
Leu Lys Ala Lys Trp Asp His Glu Arg Thr Lys Phe Ser Leu Pro Pro
285 290 295
atg atg acc ctt cta cta gga gag cca gga act gga ggg tca tcc aca 1623
Met Met Thr Leu Leu Leu Gly Glu Pro Gly Thr Gly Gly Ser Ser Thr
300 305 310
cca tca atg gta ggt gct gtg aag aag tgg cag aaa tcc gat cca caa 1671
Pro Ser Met Val Gly Ala Val Lys Lys Trp Gln Lys Ser Asp Pro Gln
315 320 325 330
cag gca cta gac acg tgg agg cag tta tca gaa gca aat tca gca ctt 1719
Gln Ala Leu Asp Thr Trp Arg Gln Leu Ser Glu Ala Asn Ser Ala Leu
335 340 345
gaa aca caa ttc act gtg cta agc aaa ttg gct gaa gaa cat cca gat 1767
Glu Thr Gln Phe Thr Val Leu Ser Lys Leu Ala Glu Glu His Pro Asp
350 355 360
gca tat aaa tct gtt gtc gag agc tgc agc atg ctt aag tca gaa aag 1815
Ala Tyr Lys Ser Val Val Glu Ser Cys Ser Met Leu Lys Ser Glu Lys
365 370 375
tgg att gag caa gct agt gac cca agc caa gct gaa gtt gtt aag gca 1863
Trp Ile Glu Gln Ala Ser Asp Pro Ser Gln Ala Glu Val Val Lys Ala
380 385 390
ttg tta ggg gca aga gac gcg atg ctt gca atc agg tct cac atg cgc 1911
Leu Leu Gly Ala Arg Asp Ala Met Leu Ala Ile Arg Ser His Met Arg
395 400 405 410
tac atg ggc aag gct gca ggc atc cca ata gaa ccc gaa tct caa act 1959
Tyr Met Gly Lys Ala Ala Gly Ile Pro Ile Glu Pro Glu Ser Gln Thr
415 420 425
cag ctc ttg gat gct act atg aat atg gag ggg gtt tta ttg gct gga 2007
Gln Leu Leu Asp Ala Thr Met Asn Met Glu Gly Val Leu Leu Ala Gly
430 435 440
gtt ccc ggt gca ggc gga ttc gac gca gtt ttt gca gtc acc tta ggt 2055
Val Pro Gly Ala Gly Gly Phe Asp Ala Val Phe Ala Val Thr Leu Gly
445 450 455
aat tca agc acc aat ttg acc aaa gca tgg agt tcg cgc aat gta tta 2103
Asn Ser Ser Thr Asn Leu Thr Lys Ala Trp Ser Ser Arg Asn Val Leu
460 465 470
gcc ttg cta gtc aga gaa gat cct cgt ggg gtt tcc tta gag agt agc 2151
Ala Leu Leu Val Arg Glu Asp Pro Arg Gly Val Ser Leu Glu Ser Ser
475 480 485 490
gat cca cga gct aag gat att tca act gcc ctc tct tcg gtt gag att 2199
Asp Pro Arg Ala Lys Asp Ile Ser Thr Ala Leu Ser Ser Val Glu Ile
495 500 505
taa taaagcttct actttgctaa catgtctaaa tagattgaaa agaccataat 2252
aagttatagc aaatgcagct tgtttgagct aaaaataaat ggttttcttg aaaatttatg 2312
taatcaaaag ttaggcagtg tttggacaac tctcatttac tctattctgt gtaacaataa 2372
gattgtgcat gttgttctct tgaaagaccg aaatgcctct tgtagcaaaa aaaaaaaaaa 2432
<210>18
<211>506
<212>PRT
<213>杜仲
<400>18
Met Ala Val Val Ala Ser Ala Pro Gly Lys Val Leu Met Thr Gly Gly
1 5 10 15
Tyr Leu Ile Leu Glu Arg Pro Asn Ala Gly Ile Val Leu Ser Thr Asn
20 25 30
Ala Arg Phe Tyr Ala Ile Val Lys Pro Leu Tyr Glu Glu Val Arg Pro
35 40 45
Asp Ser Trp Ala Trp Ala Trp Thr Asp Val Lys Leu Thr Ser Pro Gln
50 55 60
Met Ala Arg Glu Thr Met Tyr Lys Leu Ser Leu Lys His Ile Ile Thr
65 70 75 80
Gln Cys Val Ser Pro Ser Gln Ser Gly Asn Pro Phe Val Glu His Ala
85 90 95
Val Gln Tyr Ala Val Ala Ala Ala His Ala Ile Phe Asp Asn Asn Lys
100 105 110
Lys Asp Ala Leu Gln Lys Arg Leu Leu Gln Gly Leu Asp Ile Thr Ile
115 120 125
Leu Gly Cys Asn Asp Phe Tyr Ser Tyr Arg Gly Glu Ile Glu Ala Arg
130 135 140
Gly Leu Pro Leu Thr Pro Asp Ser Leu Ala Ser Leu Pro Pro Phe Ser
145 150 155 160
Ser Ile Thr Phe Asn Ala Glu Glu Ser Ser Glu Lys Asn Cys Lys Pro
165 170 175
Glu Val Ala Lys Thr Gly Leu Gly Ser Ser Ala Ala Met Thr Thr Ala
180 185 190
Val Val Ala Ala Leu Leu His Tyr Leu Glu Val Val Asp Leu Ser Ser
195 200 205
Leu Thr Glu Ala Gln Asn Gln Asn Val Glu Gln Asp Leu Asp Leu Val
210 215 220
His Val Ile Ala Gln Thr Ala His Cys Ile Ala Gln Gly Lys Val Gly
225 230 235 240
Ser Gly Phe Asp Val Ser Ser Ala Val Tyr Gly Ser Gln His Tyr Val
245 250 255
Arg Phe Ser Pro Asp Val Leu Ser Ser Ala Gln Gly Ala Val Lys Gly
260 265 270
Met Pro Leu Glu Glu Val Ile Ser Asp Val Leu Lys Ala Lys Trp Asp
275 280 285
His Glu Arg Thr Lys Phe Ser Leu Pro Pro Met Met Thr Leu Leu Leu
290 295 300
Gly Glu Pro Gly Thr Gly Gly Ser Ser Thr Pro Ser Met Val Gly Ala
305 310 315 320
Val Lys Lys Trp Gln Lys Ser Asp Pro Gln Gln Ala Leu Asp Thr Trp
325 330 335
Arg Gln Leu Ser Glu Ala Asn Ser Ala Leu Glu Thr Gln Phe Thr Val
340 345 350
Leu Ser Lys Leu Ala Glu Glu His Pro Asp Ala Tyr Lys Ser Val Val
355 360 365
Glu Ser Cys Ser Met Leu Lys Ser Glu Lys Trp Ile Glu Gln Ala Ser
370 375 380
Asp Pro Ser Gln Ala Glu Val Val Lys Ala Leu Leu Gly Ala Arg Asp
385 390 395 400
Ala Met Leu Ala Ile Arg Ser His Met Arg Tyr Met Gly Lys Ala Ala
405 410 415
Gly Ile Pro Ile Glu Pro Glu Ser Gln Thr Gln Leu Leu Asp Ala Thr
420 425 430
Met Asn Met Glu Gly Val Leu Leu Ala Gly Val Pro Gly Ala Gly Gly
435 440 445
Phe Asp Ala Val Phe Ala Val Thr Leu Gly Asn Ser Ser Thr Asn Leu
450 455 460
Thr Lys Ala Trp Ser Ser Arg Asn Val Leu Ala Leu Leu Val Arg Glu
465 470 475 480
Asp Pro Arg Gly Val Ser Leu Glu Ser Ser Asp Pro Arg Ala Lys Asp
485 490 495
Ile Ser Thr Ala Leu Ser Ser Val Glu Ile
500 505
<210>19
<211>1542
<212>DNA
<213>杜仲
<220>
<221>CDS
<222>(68)..(1324)
<223>甲羟戊酸焦磷酸脱酸酶
<400>19
ggatctgctt ctcctacttt ctctctctag ggaattcttc agttttttca tttttttggg 60
gggagaa atg gcc gat gaa tca cag aaa tgg gtg ttg acg gtg acg gcg 109
Met Ala Asp Glu Ser Gln Lys Trp Val Leu Thr Val Thr Ala
1 5 10
cag acg ccg acg aac ata gca gtg atc aag tat tgg ggg aag agg gat 157
Gln Thr Pro Thr Asn Ile Ala Val Ile Lys Tyr Trp Gly Lys Arg Asp
15 20 25 30
gaa acc cta att ttg ccg atc aat gac agt atc agt gtc act cta gat 205
Glu Thr Leu Ile Leu Pro Ile Asn Asp Ser Ile Ser Val Thr Leu Asp
35 40 45
cca gaa cac ttg tgt act acc acc acc gtt gcc gtg agt ccg agc ttc 253
Pro Glu His Leu Cys Thr Thr Thr Thr Val Ala Val Ser Pro Ser Phe
50 55 60
gat caa gat cgc atg tgg ctc aat ggc aag gaa atc tct ctt tct gga 301
Asp Gln Asp Arg Met Trp Leu Asn Gly Lys Glu Ile Ser Leu Ser Gly
65 70 75
ggt aga tat caa aat tgt ttg agg gaa att cga gct cga gct agt gat 349
Gly Arg Tyr Gln Asn Cys Leu Arg Glu Ile Arg Ala Arg Ala Ser Asp
80 85 90
gtt gag gac gag aaa aag ggt att aaa atc acg aaa aaa gat tgg gag 397
Val Glu Asp Glu Lys Lys Gly Ile Lys Ile Thr Lys Lys Asp Trp Glu
95 100 105 110
aaa ttg cat ttg cat gtt gct tct tat aac aat ttc cca acg gct gct 445
Lys Leu His Leu His Val Ala Ser Tyr Asn Asn Phe Pro Thr Ala Ala
115 120 125
ggt ttg gct tcc tca gct gcc ggt ttt gct tgt ctt gtt ttt gcg ctt 493
Gly Leu Ala Ser Ser Ala Ala Gly Phe Ala Cys Leu Val Phe Ala Leu
130 135 140
gca aag ttt atg aac gtc cag gaa gat caa agc aaa ctt tcg gct ata 541
Ala Lys Phe Met Asn Val Gln Glu Asp Gln Ser Lys Leu Ser Ala Ile
145 150 155
gcg agg caa ggt tca ggc agc gct tgt cgc agt cta ttt ggc gga ttt 589
Ala Arg Gln Gly Ser Gly Ser Ala Cys Arg Ser Leu Phe Gly Gly Phe
160 165 170
gtg aaa tgg atc atg gga aaa caa gag gat gga agt gac agc atc gcc 637
Val Lys Trp Ile Met Gly Lys Gln Glu Asp Gly Ser Asp Ser Ile Ala
175 180 185 190
gtt caa tta aca gat gag aaa cac tgg gat gat ctt gtt att gtt atc 685
Val Gln Leu Thr Asp Glu Lys His Trp Asp Asp Leu Val Ile Val Ile
195 200 205
gct gtg gta agt tca cga cag aag gaa aca agt agc act tca gga atg 733
Ala Val Val Ser Ser Arg Gln Lys Glu Thr Ser Ser Thr Ser Gly Met
210 215 220
cgc gag agt gtc gaa acg agt aag ctc ata caa cat aga gca gag gaa 781
Arg Glu Ser Val Glu Thr Ser Lys Leu Ile Gln His Arg Ala Glu Glu
225 230 235
gta gta cca aaa cgc ata att caa atg gaa gaa gcc att aaa aat cgc 829
Val Val Pro Lys Arg Ile Ile Gln Met Glu Glu Ala Ile Lys Asn Arg
240 245 250
gat ttt cct tct ttt gca aac ctc gcc tgt gct gac agc aat cag ttt 877
Asp Phe Pro Ser Phe Ala Asn Leu Ala Cys Ala Asp Ser Asn Gln Phe
255 260 265 270
cac gct gtc tgc ctc gac aca tct cct cct att ttc tac atg aac gat 925
His Ala Val Cys Leu Asp Thr Ser Pro Pro Ile Phe Tyr Met Asn Asp
275 280 285
acc tct cat agg att att agc tat gtt gag aaa tgg aac cgt tct gaa 973
Thr Ser His Arg Ile Ile Ser Tyr Val Glu Lys Trp Asn Arg Ser Glu
290 295 300
gga acc cct cag gtt gct tat act ttt gat gct ggg cca aat gcc gtc 1021
Gly Thr Pro Gln Val Ala Tyr Thr Phe Asp Ala Gly Pro Asn Ala Val
305 310 315
cta atc gca cgc aat aga aaa acc gct gct ctc ttg ctc caa agg ttg 1069
Leu Ile Ala Arg Asn Arg Lys Thr Ala Ala Leu Leu Leu Gln Arg Leu
320 325 330
ctc tat tac ttc ccc cca caa tcg gat acc gat ttg aac agt tat gtt 1117
Leu Tyr Tyr Phe Pro Pro Gln Ser Asp Thr Asp Leu Asn Ser Tyr Val
335 340 345 350
atc ggc gat aag tcg att tta caa gat gtc ggg gtt aag gac atg aaa 1165
Ile Gly Asp Lys Ser Ile Leu Gln Asp Val Gly Val Lys Asp Met Lys
355 360 365
gat gtg gaa gct ctg cct tca cct cca gaa ata aaa gat gcc caa aaa 1213
Asp Val Glu Ala Leu Pro Ser Pro Pro Glu Ile Lys Asp Ala Gln Lys
370 375 380
tac aag gga gat gtt agt tat ttc ata tgc aca aga cct ggg aaa ggc 1261
Tyr Lys Gly Asp Val Ser Tyr Phe Ile Cys Thr Arg Pro Gly Lys Gly
385 390 395
ccg gtt tta ctc gcc gac gaa agc caa gct ctt ctt aac cct aaa acc 1309
Pro Val Leu Leu Ala Asp Glu Ser Gln Ala Leu Leu Asn Pro Lys Thr
400 405 410
ggg ctg cca aaa taa ataatgaaag gagaatactc aatttataag cttttgtaat 1364
Gly Leu Pro Lys
415
gctttgtttt caagaaggat ttgcttacag atgatgtttg taacctttgg aaacttgatc 1424
ttttgttttt agggtttgcc attaagttgg atcttctttg tactacccat tttgtggatg 1484
aataatttga tcttgaacca ataaccattt aactttgttg attcgaaaaa aaaaaaaa 1542
<210>20
<211>418
<212>PRT
<213>杜仲
<400>20
Met Ala Asp Glu Ser Gln Lys Trp Val Leu Thr Val Thr Ala Gln Thr
1 5 10 15
Pro Thr Asn Ile Ala Val Ile Lys Tyr Trp Gly Lys Arg Asp Glu Thr
20 25 30
Leu Ile Leu Pro Ile Asn Asp Ser Ile Ser Val Thr Leu Asp Pro Glu
35 40 45
His Leu Cys Thr Thr Thr Thr Val Ala Val Ser Pro Ser Phe Asp Gln
50 55 60
Asp Arg Met Trp Leu Asn Gly Lys Glu Ile Ser Leu Ser Gly Gly Arg
65 70 75 80
Tyr Gln Asn Cys Leu Arg Glu Ile Arg Ala Arg Ala Ser Asp Val Glu
85 90 95
Asp Glu Lys Lys Gly Ile Lys Ile Thr Lys Lys Asp Trp Glu Lys Leu
100 105 110
His Leu His Val Ala Ser Tyr Asn Asn Phe Pro Thr Ala Ala Gly Leu
115 120 125
Ala Ser Ser Ala Ala Gly Phe Ala Cys Leu Val Phe Ala Leu Ala Lys
130 135 140
Phe Met Asn Val Gln Glu Asp Gln Ser Lys Leu Ser Ala Ile Ala Arg
145 150 155 160
Gln Gly Ser Gly Ser Ala Cys Arg Ser Leu Phe Gly Gly Phe Val Lys
165 170 175
Trp Ile Met Gly Lys Gln Glu Asp Gly Ser Asp Ser Ile Ala Val Gln
180 185 190
Leu Thr Asp Glu Lys His Trp Asp Asp Leu Val Ile Val Ile Ala Val
195 200 205
Val Ser Ser Arg Gln Lys Glu Thr Ser Ser Thr Ser Gly Met Arg Glu
210 215 220
Ser Val Glu Thr Ser Lys Leu Ile Gln His Arg Ala Glu Glu Val Val
225 230 235 240
Pro Lys Arg Ile Ile Gln Met Glu Glu Ala Ile Lys Asn Arg Asp Phe
245 250 255
Pro Ser Phe Ala Asn Leu Ala Cys Ala Asp Ser Asn Gln Phe His Ala
260 265 270
Val Cys Leu Asp Thr Ser Pro Pro Ile Phe Tyr Met Asn Asp Thr Ser
275 280 285
His Arg Ile Ile Ser Tyr Val Glu Lys Trp Asn Arg Ser Glu Gly Thr
290 295 300
Pro Gln Val Ala Tyr Thr Phe Asp Ala Gly Pro Asn Ala Val Leu Ile
305 310 315 320
Ala Arg Asn Arg Lys Thr Ala Ala Leu Leu Leu Gln Arg Leu Leu Tyr
325 330 335
Tyr Phe Pro Pro Gln Ser Asp Thr Asp Leu Asn Ser Tyr Val Ile Gly
340 345 350
Asp Lys Ser Ile Leu Gln Asp Val Gly Val Lys Asp Met Lys Asp Val
355 360 365
Glu Ala Leu Pro Ser Pro Pro Glu Ile Lys Asp Ala Gln Lys Tyr Lys
370 375 380
Gly Asp Val Ser Tyr Phe Ile Cys Thr Arg Pro Gly Lys Gly Pro Val
385 390 395 400
Leu Leu Ala Asp Glu Ser Gln Ala Leu Leu Asn Pro Lys Thr Gly Leu
405 410 415
Pro Lys
<210>21
<211>22
<212>DNA
<213>人工
<220>
<223>N192-82-tree_1968_3R_s1引物
<400>21
cattaccagc caccaagctt gg 22
<210>22
<211>23
<212>DNA
<213>人工
<220>
<223>N192-82-tree_1968_3R_s2引物
<400>22
ccaagtacct tgcagaagca agg 23
<210>23
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_1968_s引物
<400>23
tagcagctgt aatacgactc actataggga atattaagct accggaggtt ttaggtgctc 60
<210>24
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_1968_a引物
<400>24
gtaagcgtga cataactaat tacatgatgc ggccctctag tcaataggta gggattagac 60
<210>25
<211>23
<212>DNA
<213>人工
<220>
<223>N192-84-tree_11012_3R_s1引物
<400>25
ccaatgggtg gctttcttgg aac 23
<210>26
<211>22
<212>DNA
<213>人工
<220>
<223>N192-84-tree_11012_3R_s2引物
<400>26
tgacgttgta gtagcaggtg gc 22
<210>27
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_11012_s引物
<400>27
tagcagctgt aatacgactc actataggga atattaagct tcactgtgag caattaactg 60
<210>28
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_11012_a引物
<400>28
gtaagcgtga cataactaat tacatgatgc ggccctctag tgcgattaca gaagctcaac 60
<210>29
<211>22
<212>DNA
<213>人工
<220>
<223>N192-71-tree_6098_3R_s1引物
<400>29
ggtcgtcttg aagtgggaag tg 22
<210>30
<211>22
<212>DNA
<213>人工
<220>
<223>N192-71-tree_6098_3R_s2引物
<400>30
tgggtggaga gtagttcatg gg 22
<210>31
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_6098_s引物
<400>31
tagcagctgt aatacgactc actataggga atattaagct tgtttgttct cgagaatccg 60
<210>32
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_6098_a引物
<400>32
gtaagcgtga cataactaat tacatgatgc ggccctctag aatgccagtt ttggtcagtc 60
<210>33
<211>22
<212>DNA
<213>人工
<220>
<223>N192-36-tree_10370_3R_s1引物
<400>33
ctgtgtccag caggaagcaa tg 22
<210>34
<211>22
<212>DNA
<213>人工
<220>
<223>N192-36-tree_10370_3R_s2引物
<400>34
caattgggtg gagagcagtt cg 22
<210>35
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_10370_s引物
<400>35
tagcagctgt aatacgactc actataggga atattaagct atccgttcgt ttctcgtgag 60
<210>36
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_10370_a引物
<400>36
gtaagcgtga cataactaat tacatgatgc ggccctctag ttttctaccc agaatcaccg 60
<210>37
<211>23
<212>DNA
<213>人工
<220>
<223>N192-45-tree_8220_5R_a1引物
<400>37
ggggtttgag ggcatatgaa tgc 23
<210>38
<211>22
<212>DNA
<213>人工
<220>
<223>N192-45-tree_8220_5R_a2引物
<400>38
cgtcggacgt ccattgattt cg 22
<210>39
<211>22
<212>DNA
<213>人工
<220>
<223>N192-45-tree_8220_5R_3R_s1引物
<400>39
atccgtaagg atgacgacgg tc 22
<210>40
<211>22
<212>DNA
<213>人工
<220>
<223>N192-45-tree_8220_5R_3R_s2引物
<400>40
tccacctctc gtctcatctg ca 22
<210>41
<211>15
<212>DNA
<213>人工
<220>
<223>N192-45-tree_8220_5R_p引物
<400>41
cctttgcaat gcttc 15
<210>42
<211>22
<212>DNA
<213>人工
<220>
<223>N192-47-tree_13453_5R_a1引物
<400>42
ttgcggatct tttcacgcca gc 22
<210>43
<211>22
<212>DNA
<213>人工
<220>
<223>N192-47-tree_13453_5R_a2引物
<400>43
tatgtggctt gagagagtcg cc 22
<210>44
<211>22
<212>DNA
<213>人工
<220>
<223>N192-47-tree_13453_5R_3R_s1引物
<400>44
attgcaccca aggcgttggt tc 22
<210>45
<211>22
<212>DNA
<213>人工
<220>
<223>N192-47-tree_13453_5R_3R_s2引物
<400>45
atcaaatcag tggtggccgg ga 22
<210>46
<211>14
<212>DNA
<213>人工
<220>
<223>N192-47-tree_13453_5R_p引物
<400>46
ttccggtgat tctc 14
<210>47
<211>22
<212>DNA
<213>人工
<220>
<223>tree 1729引物
<400>47
cacctacctc atgttcactg gg 22
<210>48
<211>23
<212>DNA
<213>人工
<220>
<223>tree 1729引物套
<400>48
gaggcaaaca ttggacttgc tgc 23
<210>49
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_1729_s引物
<400>49
tagcagctgt aatacgactc actataggga atattaagct ggggcggaga cgaatggagg 60
<210>50
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_1729_a引物
<400>50
gtaagcgtga cataactaat tacatgatgc ggccctctag atcaagacga gccgccaaag 60
<210>51
<211>23
<212>DNA
<213>人工
<220>
<223>S_No.243_P.10_1引物
<220>
<221>misc_feature
<222>(21)..(21)
<223>n为a、t、c或g中任一个。
<220>
<221>misc_feature
<222>(6)..(6)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(9)..(9)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(12)..(12)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(15)..(15)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(18)..(18)
<223>n为肌苷。
<400>51
aaracnggny tnggnwsnws ngc 23
<210>52
<211>20
<212>DNA
<213>人工
<220>
<223>AS_No.243_P.10_1引物
<220>
<221>misc_feature
<222>(9)..(9)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(12)..(12)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(15)..(15)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(18)..(18)
<223>n为a、c、t或g中任一个。
<400>52
gcrtcrwanc cnccngcncc 20
<210>53
<211>28
<212>DNA
<213>人工
<220>
<223>S_No.243_P.10_2引物
<220>
<221>misc_feature
<222>(6)..(6)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(12)..(12)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(15)..(15)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(18)..(18)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(21)..(21)
<223>n为肌苷。
<400>53
carggnaarr tnggnwsngg nttygayg 28
<210>54
<211>20
<212>DNA
<213>人工
<220>
<223>AS_No.243_P.10_2引物
<220>
<221>misc_feature
<222>(6)..(6)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(9)..(9)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(12)..(12)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(15)..(15)
<223>n为肌苷。
<220>
<221>misc_feature
<222>(18)..(18)
<223>n为a、c、t或g中任一个。
<400>54
tcrwanccnc cngcnccngg 20
<210>55
<211>22
<212>DNA
<213>人工
<220>
<223>No.243_P.51_race_a2引物
<400>55
cccacttggc tttcaggaca tc 22
<210>56
<211>22
<212>DNA
<213>人工
<220>
<223>No.243_P.51_race_a1引物
<400>56
gcacctacca ttgatggtgt gg 22
<210>57
<211>22
<212>DNA
<213>人工
<220>
<223>No.243_P.51_race_s1引物
<400>57
atctgttgtc gagagctgca gc 22
<210>58
<211>22
<212>DNA
<213>人工
<220>
<223>No.243_P.51_race_s2引物
<400>58
agacgcgatg cttgcaatca gg 22
<210>59
<211>15
<212>DNA
<213>人工
<220>
<223>No.243_P.51_race_p引物
<400>59
gcagccttgc ccatg 15
<210>60
<211>26
<212>DNA
<213>人工
<220>
<223>cDNA文库筛选用有义引物no.243
<400>60
acctgacgta ctttcttctg ctcagg 26
<210>61
<211>26
<212>DNA
<213>人工
<220>
<223>cDNA文库筛选用反义引物no.243
<400>61
tccaagagct gagtttgaga ttcggg 26
<210>62
<211>60
<212>DNA
<213>人工
<220>
<223>互补性试验用有义引物no.243
<400>62
tagcagctgt aatacgactc actataggga atattaagct accggattcg gttgtattcc 60
<210>63
<211>60
<212>DNA
<213>人工
<220>
<223>互补性试验用反义引物no.243
<400>63
gtaagcgtga cataactaat tacatgatgc ggccctctag agagttgtcc aaacactgcc 60
<210>64
<211>23
<212>DNA
<213>人工
<220>
<223>N192-102-tree_13103_3R_s1引物
<400>64
gatcaagtat tgggggaaga ggg 23
<210>65
<211>22
<212>DNA
<213>人工
<220>
<223>N192-102-tree_13103_3R_s2引物
<400>65
cgagctagtg atgttgagga cg 22
<210>66
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_13103_s引物
<400>66
tagcagctgt aatacgactc actataggga atattaagct agaaatggcc gatgaatcac 60
<210>67
<211>60
<212>DNA
<213>人工
<220>
<223>N219-36-tree_13103_a引物
<400>67
gtaagcgtga cataactaat tacatgatgc ggccctctag aaattgagta ttctcctttc 60
60
Claims (3)
1.基因，该基因是参与由杜仲的甲羟戊酸合成异戊烯焦磷酸的基因，该基因编码由SEQ ID NO.2的氨基酸序列中第1位-408位的氨基酸、或SEQ ID NO.4的氨基酸序列中第1位-408位的氨基酸组成的乙酰-CoA C-乙酰转移酶。
2.如权利要求1所述的基因，其中，所述基因为含有SEQ IDNO.1的碱基序列中第101位-1327位的核苷酸的基因、或含有SEQ IDNO.3的碱基序列中第128位-1354位核苷酸的基因。
3.如权利要求2所述的基因，其中，所述述基因为含有SEQ IDNO.1的碱基序列中第1位-1516位核苷酸的基因、或含有SEQ IDNO.3的碱基序列中第1位-1757位核苷酸的基因。
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2007163872 | 2007-06-21 | ||
| JP2007-163872 | 2007-06-21 | ||
| JP2007163872A JP2009000046A (ja) | 2007-06-21 | 2007-06-21 | トチュウのメバロン酸経路の酵素をコードする遺伝子 |
Related Child Applications (5)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110358647.8A Division CN102505018B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN201110358667.5A Division CN102505019B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN201110358587XA Division CN102505017A (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN2011103585653A Division CN102492701B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN201110355190.5A Division CN102453724B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101328485A CN101328485A (zh) | 2008-12-24 |
| CN101328485B true CN101328485B (zh) | 2012-02-15 |
Family
ID=40204494
Family Applications (6)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110355190.5A Expired - Fee Related CN102453724B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN201110358667.5A Expired - Fee Related CN102505019B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN201110358587XA Pending CN102505017A (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN2008101253609A Expired - Fee Related CN101328485B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN2011103585653A Expired - Fee Related CN102492701B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN201110358647.8A Expired - Fee Related CN102505018B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
Family Applications Before (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110355190.5A Expired - Fee Related CN102453724B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN201110358667.5A Expired - Fee Related CN102505019B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN201110358587XA Pending CN102505017A (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
Family Applications After (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011103585653A Expired - Fee Related CN102492701B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
| CN201110358647.8A Expired - Fee Related CN102505018B (zh) | 2007-06-21 | 2008-06-20 | 编码杜仲的甲羟戊酸途径的酶的基因 |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JP2009000046A (zh) |
| CN (6) | CN102453724B (zh) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150132813A1 (en) * | 2012-05-11 | 2015-05-14 | Regents Of The University Of Minnesota | Biosynthetic pathways, recombinant cells, and methods |
| CN103571857B (zh) * | 2012-07-27 | 2016-03-23 | 中国中医科学院中药研究所 | 芍药乙酰CoA酰基转移酶(PLAACT)基因及其编码产物和应用 |
| JP5870464B2 (ja) * | 2014-08-12 | 2016-03-01 | 日立造船株式会社 | 長鎖トランス型プレニル二リン酸合成酵素遺伝子 |
| CN105039369A (zh) * | 2015-09-02 | 2015-11-11 | 江西中医药大学 | 穿心莲甲羟戊酸 5-焦磷酸脱羧酶基因 |
| JP6090700B2 (ja) * | 2015-12-25 | 2017-03-08 | 日立造船株式会社 | 長鎖トランス型プレニル二リン酸合成酵素遺伝子 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1531594A (zh) * | 2000-10-30 | 2004-09-22 | �����ɷ� | 新的草甘膦n-乙酰转移酶(gat)基因 |
| CN1772908A (zh) * | 2005-10-17 | 2006-05-17 | 中国农业科学院生物技术研究所 | 草甘膦乙酰转移酶基因及其应用 |
| CN1806042A (zh) * | 2003-06-12 | 2006-07-19 | 帝斯曼知识产权资产管理有限公司 | 具有反馈抗性的甲羟戊酸激酶 |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI250210B (en) * | 1998-05-06 | 2006-03-01 | Dsm Ip Assets Bv | An isolated DNA sequence coding for an enzyme involved in the mevalonate pathway or the pathway from isopentenyl pyrophosphate to farnesyl pyrophosphate |
| US6916972B2 (en) * | 1998-11-05 | 2005-07-12 | E. I. Du Pont De Nemours And Company | Mevalonate synthesis enzymes |
| CA2449885A1 (en) * | 2001-06-06 | 2002-12-12 | Roche Vitamins Ag | Improved isoprenoid production |
| CN100415882C (zh) * | 2003-10-28 | 2008-09-03 | 北京农业生物技术研究中心 | 一种调控毛白杨中木质素的方法 |
| WO2005071072A1 (fr) * | 2004-01-13 | 2005-08-04 | The Institute Of Biophysics Chinese Academy Of Sciences | Transcriptase inverse du virus de la leucemie murine recombinee, gene codant pour ladite transcriptase inverse et methode permettant d'exprimer ladite transcriptase inverse |
| KR100704776B1 (ko) * | 2004-11-06 | 2007-04-10 | 경상대학교산학협력단 | 내재적 비메발론산 경로는 불활성화되어 있고, 외래메발론산 경로에 관련되는 모든 효소를 코딩하는 ｄｎａ로형질전환된 숙주세포를 이용한 메발론산 경로 및비메발론산 경로 중 어느 하나 또는 모두를 저해하는물질을 탐색하는 방법 |
| US8338155B2 (en) * | 2004-12-14 | 2012-12-25 | Dsm Ip Assets B.V. | Modified mevalonate kinase with reduced feedback inhibition |
| ES2546484T3 (es) * | 2005-03-18 | 2015-09-24 | Dsm Ip Assets B.V. | Producción de carotenoides en levadura y hongos oleaginosos |
-
2007
- 2007-06-21 JP JP2007163872A patent/JP2009000046A/ja active Pending
-
2008
- 2008-06-20 CN CN201110355190.5A patent/CN102453724B/zh not_active Expired - Fee Related
- 2008-06-20 CN CN201110358667.5A patent/CN102505019B/zh not_active Expired - Fee Related
- 2008-06-20 CN CN201110358587XA patent/CN102505017A/zh active Pending
- 2008-06-20 CN CN2008101253609A patent/CN101328485B/zh not_active Expired - Fee Related
- 2008-06-20 CN CN2011103585653A patent/CN102492701B/zh not_active Expired - Fee Related
- 2008-06-20 CN CN201110358647.8A patent/CN102505018B/zh not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1531594A (zh) * | 2000-10-30 | 2004-09-22 | �����ɷ� | 新的草甘膦n-乙酰转移酶(gat)基因 |
| CN1806042A (zh) * | 2003-06-12 | 2006-07-19 | 帝斯曼知识产权资产管理有限公司 | 具有反馈抗性的甲羟戊酸激酶 |
| CN1772908A (zh) * | 2005-10-17 | 2006-05-17 | 中国农业科学院生物技术研究所 | 草甘膦乙酰转移酶基因及其应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102505017A (zh) | 2012-06-20 |
| CN102505018B (zh) | 2014-06-04 |
| CN102505019B (zh) | 2015-03-11 |
| CN102505019A (zh) | 2012-06-20 |
| CN102492701A (zh) | 2012-06-13 |
| JP2009000046A (ja) | 2009-01-08 |
| CN102453724B (zh) | 2014-01-01 |
| CN102453724A (zh) | 2012-05-16 |
| CN102505018A (zh) | 2012-06-20 |
| CN102492701B (zh) | 2013-11-06 |
| CN101328485A (zh) | 2008-12-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU763068B2 (en) | Nucleic acids encoding (taxus) geranylgeranyl diphosphate synthase, and methods of use | |
| CN101328485B (zh) | 编码杜仲的甲羟戊酸途径的酶的基因 | |
| CN102124111B (zh) | 参与生物合成的新基因 | |
| EP3482627A1 (en) | Endophytes and related methods | |
| CN101182544A (zh) | 转ads基因提高青蒿中青蒿素含量的方法 | |
| CN110892074A (zh) | 用于增加香蕉的保质期的组成物及方法 | |
| CN112424364A (zh) | 减少咖啡豆中咖啡因含量的组合物及方法 | |
| Nitschke et al. | Complementation of the cs dis2‐11 cell cycle mutant of Schizosaccharomyces pombe by a protein phosphatase from Arabidopsis thaliana. | |
| CN111386342A (zh) | 具有蓝色系花色的转化植物及其制作方法 | |
| CN101687907A (zh) | 转移酶、差向异构酶、编码它们的多核苷酸及其用途 | |
| Schardl et al. | Molecular biology and evolution of the grass endophytes | |
| CN110592114B (zh) | 水稻生长素糖基转移酶基因的应用 | |
| HU221515B (en) | Transgenic plants with improved biomass production | |
| US8476491B2 (en) | Method of utilizing the PTS gene and RNA interference of the ads gene to increase patchouli alcohol content in Artemisia annua L. | |
| CN101663393A (zh) | 天冬酰胺和蛋白质增强的玉米植物和种子 | |
| CN103052709A (zh) | 原伊鲁烯合酶 | |
| CN103649318B (zh) | 通过改变激酶和磷酸酶的表达水平加快植物生长并提高产量的方法 | |
| US20030148489A1 (en) | Farnesyl pyrophoshate synthase protein, nucleic acid and promoter region thereof | |
| JP2013226153A (ja) | トチュウのメバロン酸経路の酵素をコードする遺伝子 | |
| AU2019200890B2 (en) | Novel endophytes (3) | |
| CN106868028A (zh) | 粳稻的als突变型基因及其蛋白和应用 | |
| AU2021202922A1 (en) | Designer endophytes (4) | |
| CN110628809A (zh) | 一种利用AaTGA6基因提高青蒿中青蒿素含量的方法 | |
| Jumali et al. | Molecular cloning and characterization of strictosidine synthase, a key gene in biosynthesis of mitragynine from Mitragyna speciosa | |
| CN114457095A (zh) | 水稻丙酮酸激酶基因OsPK5的基因工程应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120215 Termination date: 20200620 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |