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1. 发明申请

WO2017081520A1 PRODUCTION OF IN VIVO N-DEGLYCOSYLATED RECOMBINANT PROTEINS BY CO-EXPRESSION WITH ENDO H 审中-公开
标题翻译：与ENDO H共同生产体内N-脱酰基重组蛋白质
公开(公告)号：WO2017081520A1
公开(公告)日：2017-05-18
申请号：PCT/IB2015/058781
申请日：2015-11-13
申请人： MAMMEDOV, Tarlan
发明人： MAMMEDOV, Tarlan
IPC分类号： C12N9/14 , C07K14/36 , C12N15/82
CPC分类号： C12N15/8242 , C12N9/2402
摘要： Plants have emerged as an alternative expression system and are increasingly being used byindustry and academia for producing target proteins. However, the ability of plants to glycosylate proteins can be a significant limitation for those proteins, which do not require N- glycosylation. For example, Plasmodium falciparum proteins, or A chain of human factor XIIIdo not carry N-linked glycans, or the protective antigen (PA) of Bacillus anthracisis not a glycoprotein; however, these proteins contain potential N-linked glycosylation sites that can be aberrantly glycosylated during expression in yeast, mammalian, or plant systems, potentially leading toreduced functionality and immunogenicity because of incorrect⁄altered folding and⁄or masking of epitopes. To overcome this problem we have recently developed a strategy of enzymatic deglycosylation of proteins in vivo by co-expressing with bacterial PNGase F (Peptide: N-glycosidase F) using transient expression in plants (WIPO Patent Application WO/2012/170678), which allowed production of malaria vaccine candidate Pfs48/45, which can provide ahigh transmission blocking (TB) activity (Mamedov et al., 2012). In addition, other deglycosylated antigens induced significantly higher levels of toxin-neutralizing antibody responses in mice than compared with glycosylated forms (Mamedov et al, manuscript has been submitted). Although a PNGase F treatment (in vivodeglycosylation) removes the oligosaccharide intact, but causes amino acid change in the deglycosylated protein due to deamidation of the asparagine (N) in the NxS/T site (sequence) into an aspartate (D). In this study, a strategy was developed for production of target proteins in plants in non-N-glycosylated form, but with no amino acid change in the NxS/T site of the resulting deglycosylated proteins, which can provide production of non-N- glycosylated recombinant proteins in plants or other eukaryotic system with a native-like fold. Thus, materials and methods for in vivo de- glycosylation of recombinant N-glycosylated proteins by co-expression with Endo-β-N- acetylglucosaminidase H (Endo H) in plants, using a transient expression system are described in this invention. A method of expressing active Endo Hin plants is also provided.
摘要翻译：植物已经成为替代表达系统，越来越多地被工业界和学术界用于生产靶蛋白。然而，植物对蛋白质进行糖基化的能力可能是那些不需要N-糖基化的蛋白质的显着限制。例如，恶性疟原虫蛋白或人因子XIII的A链不携带N-连接的聚糖，或者炭疽芽孢杆菌的保护性抗原（PA）不是糖蛋白; 然而，这些蛋白质含有潜在的N-连接糖基化位点，其在酵母，哺乳动物或植物系统中的表达过程中可能异常糖基化，由于不正确的/改变的折叠和/或掩蔽表位，潜在地导致功能性降低和免疫原性降低。为了克服这个问题，我们最近通过使用植物中的瞬时表达（WIPO专利申请WO / 2012/170678）与细菌PNGase F（肽：N-糖苷酶F）共表达开发了体内蛋白质酶促去糖基化的策略（WIPO专利申请WO / 2012/170678）允许生产可提供高传输阻断（TB）活性的疟疾疫苗候选物Pfs48 / 45（Mamedov等，2012）。此外，与糖基化形式相比，其他去糖基化抗原在小鼠中诱导显着更高水平的毒素中和抗体应答（Mamedov等，手稿已提交）。尽管PNGase F处理（体内糖基化）除去了完整的寡糖，但由于NxS / T位点（序列）中的天冬酰胺（N）脱酰胺成天冬氨酸（D），导致去糖基化蛋白中的氨基酸改变。在这项研究中，制定了一种策略，用于以非N-糖基化形式在植物中产生目标蛋白质，但是所得到的去糖基化蛋白质的NxS / T位点没有氨基酸改变，这可以提供非N-糖基化在具有天然样折叠的植物或其他真核系统中糖基化重组蛋白。因此，本发明描述了使用瞬时表达系统通过在植物中与内切-β-N-乙酰氨基葡糖苷酶H（Endo H）共表达的用于体内重组N-糖基化蛋白质的糖基化的材料和方法。还提供了表达活性Endo Hin植物的方法。

2. 发明申请

WO2016183467A1 GLUCANASE PRODUCTION AND METHODS OF USING THE SAME 审中-公开
标题翻译：葡萄糖苷酶的生产及其使用方法
公开(公告)号：WO2016183467A1
公开(公告)日：2016-11-17
申请号：PCT/US2016/032418
申请日：2016-05-13
申请人： AGRIVIDA, INC.
发明人： RAAB, R. Michael , BOUGRI, Oleg , LI, Xuemei
IPC分类号： C07K14/415 , C12N15/00 , C12N15/52 , C12N9/24
CPC分类号： C12N9/24 , C12N15/8242 , C12Q1/6895 , C12Q2600/13
摘要： Methods and compositions are described for producing a glucanase in transgenic plants and then incorporating parts of the transgenic plants in animal feed. The feed glucanase enzyme displays activity across a broad pH range, and tolerance to temperatures that are often encountered during the process of preparing animal feeds. Producing the enzyme in the transgenic plant enhances the thermal stability of the enzyme.
摘要翻译：描述了用于在转基因植物中产生葡聚糖酶的方法和组合物，然后将部分转基因植物掺入动物饲料中。饲料葡聚糖酶显示在宽的pH范围内的活性，以及在制备动物饲料的过程中经常遇到的温度的耐受性。在转基因植物中产生酶增强酶的热稳定性。

3. 发明申请

WO2016070206A3 POLYPEPTIDE FOR THE ENZYMATIC DETOXIFICATION OF ZEARALENONE, ISOLATED POLYNUCLEOTIDE, AND ASSOCIATED ADDITIVE, USE AND METHOD 审中-公开
标题翻译：多肽，其用于玉米赤霉烯酮和孤立的多核苷酸和其它材料，用途和方法相同的酶法解毒
公开(公告)号：WO2016070206A3
公开(公告)日：2016-07-21
申请号：PCT/AT2015000138
申请日：2015-11-04
申请人： ERBER AG
发明人： TORRES ACOSTA JEAN ANTONIO , ADAM GERHARD , KUNZ-VEKIRU ELISAVET , MOLL WULF-DIETER , MITTERBAUER RUDOLF , SCHMEITZL CLEMENS
IPC分类号： C12N9/02 , C12N15/82
CPC分类号： C12N9/0073 , C12N15/82 , C12N15/8242
摘要： The invention relates to a polypeptide for the enzymatic detoxification of zearalenone, said polypeptide being a monooxygenase which converts the keto group in position 7 of zearalenone into an ester group, the monooxygenase in particular being an amino acid sequence selected from the group comprising sequence ID No. 1, 2 and 3 or a functional variant thereof. The functional variant and at least one of the amino acid sequences has a sequence identity of at least 60%, preferably at least 70%, more preferably at least 80% and most preferably 90%.
摘要翻译：对于多肽玉米赤霉烯酮的酶促解毒的多肽是在选自序列ID号的基团转化为酯基特别是单加氧酶在玉米赤霉烯酮的7位酮基。 1，2和3或功能性变体选择的氨基酸序列，其中所述功能性变体和氨基酸序列至少60％的至少一种，优选至少70％，更优选至少80％，并且最优选由至少90％序列同一性的之间。

4. 发明申请

WO2016016208A1 SOLANUM LYCOPERSICUM PLANTS HAVING PINK GLOSSY FRUITS 审中-公开
标题翻译：具有粉红色水果水果的SOLANUM LYCOPERSICUM植物
公开(公告)号：WO2016016208A1
公开(公告)日：2016-02-04
申请号：PCT/EP2015/067200
申请日：2015-07-28
申请人： NUNHEMS B.V.
发明人： VRIEZEN, Hendrik Willem , VERBAKEL, Henk
IPC分类号： A01H5/08 , C12N15/82
CPC分类号： A01H5/08 , A01H1/02 , C12N15/8242 , C12N15/825
摘要： The present invention relates to cultivated plant of the species Solanum lycopersicum comprising a myb12 allele having one or more mutations, said mutations resulting in production of a mutant myb12 protein, fruits of such plants exhibiting a pink appearance and in addition comprising a mutant cuticle deficiency ( cd ) allele, whereby the fruits become glossy.
摘要翻译：本发明涉及茄科茄子的栽培植物，其包含具有一个或多个突变的myb12等位基因，所述突变导致产生突变型myb12蛋白，这种植物表现出粉红色外观的果实，并且还包含突变体角质层缺陷（ cd）等位基因，从而使果实变得光滑。

5. 发明申请

WO2015181647A1 AUGMENTATION DE LA RECOMBINAISON MÉIOTIQUE CHEZ LES PLANTES PAR INHIBITION D'UNE PROTÉINE RECQ4 OU TOP3A DU COMPLEXE RTR 审中-公开
标题翻译：通过抑制RTR复合物的RECQ4或TOP3A蛋白质在植物中的生物重建中增加
公开(公告)号：WO2015181647A1
公开(公告)日：2015-12-03
申请号：PCT/IB2015/052276
申请日：2015-03-27
申请人： INSTITUT NATIONAL DE LA RECHERCHE AGRONOMIQUE
发明人： MERCIER, Raphaël , SEGUELA-ARNAUD, Mathilde , CRISMANI, Wayne
IPC分类号： C07K14/415 , C12N15/82 , C12N9/90
CPC分类号： C12N15/8242 , C07K14/415 , C12N9/90 , C12N15/8218 , C12N15/8241 , C12Y599/01002
摘要： L'invention est relative à une méthode pour augmenter la fréquence de recombinaison méiotique chez les plantes, par Γ inhibition de la protéine RECQ4 ou TOP3A, notamment par mutagénèse ou extinction du gène RECQ4 ou TOP3A codant pour ladite protéine. La présente invention est utilisable notamment dans le domaine de l'amélioration des plantes et de la cartographie génétique.
摘要翻译：本发明涉及通过抑制RECQ4或TOP3A蛋白，特别是通过编码所述蛋白质的RECQ4或TOP3A基因的诱变或灭活来增加植物中减数分裂重组频率的方法。本发明可用于植物育种和遗传作图领域。

6. 发明申请

WO2013153237A1 QTL RESPONSIBLE FOR TOMATO FRUIT FIRMNESS 审中-公开
标题翻译： QTL对TOMATO果实的责任
公开(公告)号：WO2013153237A1
公开(公告)日：2013-10-17
申请号：PCT/EP2013/062071
申请日：2013-06-11
申请人： SYNGENTA PARTICIPATIONS AG
发明人： BAXTER, Charles , GRIVET, Laurent , BONNET, Julien , CHAPMAN, Natalie, Helene , SEYMOUR, Graham, Barron
IPC分类号： A01H5/08 , C12N15/82 , C12Q1/68 , A01H5/00
CPC分类号： G06F19/18 , A01H1/02 , A01H5/08 , C12N15/1079 , C12N15/8242 , C12Q1/6895 , C12Q2600/13
摘要： This invention relates to QTL responsible for significantly increased firmness in tomato fruit in the cultivated plant producing said tomato fruit, compared to fruit from a control tomato plant which does not have said genetic elements. A cultivated tomato plant producing tomato fruit with significantly increased fruit firmness and a method for detecting QTLs linked to significantly increased fruit firmness are also provided.
摘要翻译：本发明涉及与不具有所述遗传元件的对照番茄植物的果实相比，该QTL负责显着提高番茄果实在生产所述番茄果实的栽培植物中的坚实性。还提供了一种种植番茄果实，具有显着增加的果实硬度，以及一种检测与显着提高果实硬度相关联的QTL的方法。

7. 发明申请

WO2012095843A8 ORGANISMS WITH ALTERED STEROIDAL SAPONIN AND STEROIDAL ALKALOID LEVELS AND METHODS FOR PRODUCING SAME 审中-公开
标题翻译：具有改变的STEROIDAL SAPONIN和STEROIDAL ALKALOID水平的组织及其生产方法
公开(公告)号：WO2012095843A8
公开(公告)日：2012-10-18
申请号：PCT/IL2012000019
申请日：2012-01-12
申请人： YEDA RES & DEV , AHARONI ASAPH , ITKIN MAXIM
发明人： AHARONI ASAPH , ITKIN MAXIM
IPC分类号： C12N15/82
CPC分类号： C12N15/8242 , C07K14/415 , C12N9/0077 , C12N15/8243
摘要： The present invention relates to a key gene (designated GAME4) in the biosynthesis of steroidal saponins and steroidal (glyco) alkaloids and to means and methods for altering its expression. The invention further provides genetically modified organisms having altered levels of steroidal saponins and/or steroidal alkaloids compared to unmodified organisms. The metabolites produced by the genetically modified organisms may be further used in the pharmaceutics industry.
摘要翻译：本发明涉及甾体皂苷和类固醇（糖）生物碱的生物合成中的关键基因（称为GAME4），以及改变其表达的方法和方法。本发明进一步提供与未修饰的生物体相比具有改变的甾体皂苷和/或类固醇生物碱水平的转基因生物。由转基因生物产生的代谢物可进一步用于制药工业。

8. 发明申请

WO2012069539A1 DUAL PURPOSE POLLENIZER WATERMELONS 审中-公开
标题翻译：双用途POLLENIZER WATERMELONS
公开(公告)号：WO2012069539A1
公开(公告)日：2012-05-31
申请号：PCT/EP2011/070817
申请日：2011-11-23
申请人： NUNHEMS B. V. , DE GROOT, Erik , CHIAPPARINO, Elena
发明人： DE GROOT, Erik , CHIAPPARINO, Elena
IPC分类号： A01H5/08 , A01H5/00
CPC分类号： C12N15/8242 , A01G22/00 , A01H5/00 , A01H5/08 , A01H6/78 , A23L19/00
摘要： The application relates to the field of plant breeding, in particular watermelon breeding. Provided are diploid watermelon plants (and seeds from which these plants can be grown) which produce small, diploid, red watermelon fruits. Also provided are small, diploid watermelon fruits having an average weight of less than 1.8 kg.
摘要翻译：该应用涉及植物育种领域，特别是西瓜育种。提供二倍体西瓜植物（和这些植物可以种植的种子），其产生小的二倍体，红色西瓜水果。还提供了平均重量小于1.8kg的小型二倍体西瓜果实。

9. 发明申请

WO2011112380A3 TWO NOVEL ALKYLRESORCINOL SYNTHASE GENES FROM SORGHUM; CLONING, EXPRESSION, TRANSFORMATION AND CHARACTERIZATION 审中-公开
标题翻译：来自SORGHUM的两种新型碱性磷酸酶合成酶基因; 克隆，表达，转化和表征
公开(公告)号：WO2011112380A3
公开(公告)日：2012-03-29
申请号：PCT/US2011026466
申请日：2011-02-28
申请人： US AGRICULTURE , BAERSON SCOTT R , PAN ZHIQIANG , RIMANDO AGNES M , DAYAN FRANCK E , COOK DANIEL
发明人： BAERSON SCOTT R , PAN ZHIQIANG , RIMANDO AGNES M , DAYAN FRANCK E , COOK DANIEL
IPC分类号： C12N15/52 , A01H5/00 , C12N9/00 , C12N15/63
CPC分类号： A61K31/713 , C12N9/00 , C12N15/8242 , C12N15/825 , C12N15/8257 , C12N15/8279
摘要： Sorghum is considered to be an allelopathic crop species, producing phytotoxins such as the lipid benzoquinone sorgoleone (2-hydroxy-5-methoxy-3-[(Z,Z)-8',l l ', 14'- pentadecatrieneJ-^-benzoquinone) which likely accounts for much of its allelopathic properties. Prior investigations into the biosynthesis of sorgoleone have suggested the participation of one or more alkylresorcinol synthases (ARS), which are type III polyketide synthases (PKS) that produce 5-alkylresorcinols using medium to long-chain fatty acyl-CoA starter units via iterative condensations with malonyl-CoA. Current evidence suggests that sorgoleone biosynthesis occurs exclusively in root hair cells, involving the synthesis of a 5- pentadecatrienyl resorcinol intermediate derived from an unusual 16:3 fatty acyl-CoA starter unit. To characterize the enzymes responsible for the biosynthesis of this alkylresorcinol intermediate, a previously-described expressed sequence tag (EST) database prepared from isolated root hairs was first mined for all PKS-I ike sequences. Quantitative real-time RT- PCR analyses revealed that two of these sequences were preferentially expressed in root hairs, and recombinant enzyme studies demonstrated that both sequences (designated ARSl and ARS2) encode ARS enzymes capable of accepting a variety of fatty acyi-CoA starter units. Furthermore, RNA interference (RNAi) experiments directed against ARSl and ARS2 resulted in the generation of multiple independent transformant events exhibiting dramatically reduced sorgoleone levels. Thus, both ARSl and ARS2 are likely to participate in the biosynthesis of sorgoleone inpJanta. The sequences o?ARSl and ARS2 were also used to identify several rice genes encoding ARSs, which are likely involved in the production of defense-related alkyiresorcinols.
摘要翻译：高粱被认为是一种化感作物，产生植物毒素，如脂质苯醌山梨酮（2-羟基-5-甲氧基-3 - [（Z，Z）-8'，11'，14'-十五碳三烯基苯醌），这可能占其大部分的抗病性。对sorgoleone的生物合成的先前研究表明，一种或多种烷基间苯二酚合酶（ARS）的参与，其是使用中等至长链脂肪酰辅酶A起始单元通过迭代缩合产生5-烷基间苯二酚的III型聚酮化合物合酶（PKS）与丙二酰辅酶A。目前的证据表明，sorgoleone生物合成完全发生在根毛细胞中，涉及合成来自不寻常的16：3脂肪酰辅酶A引发剂单元的5-十五碳烯基间苯二酚中间体。为了表征负责该烷基间苯二酚中间体的生物合成的酶，首先从所有PKS-I序列开始从分离的根毛制备的先前描述的表达序列标签（EST）数据库。定量实时RT-PCR分析显示，这些序列中的两个优先在根毛中表达，重组酶研究表明两个序列（指定为ARS1和ARS2）编码能够接受多种脂肪酰辅酶A起始单位的ARS酶。此外，针对ARS1和ARS2的RNA干扰（RNAi）实验导致产生显着降低的sorgoleone水平的多个独立转化体事件。因此，ARS1和ARS2两者都可能参与了山茱萸碱的生物合成。序列o ARS1和ARS2也用于鉴定编码ARS的几个水稻基因，这些基因可能参与防御相关的烷基苯酚的生产。

10. 发明申请

WO2011112380A2 TWO NOVEL ALKYLRESORCINOL SYNTHASE GENES FROM SORGHUM; CLONING, EXPRESSION, TRANSFORMATION AND CHARACTERIZATION 审中-公开
标题翻译：来自SORGHUM的两种新型碱性磷酸酶合成酶基因; 克隆，表达，转化和表征
公开(公告)号：WO2011112380A2
公开(公告)日：2011-09-15
申请号：PCT/US2011/026466
申请日：2011-02-28
申请人： THE UNITED STATES OF AMERICA, AS REPRESENTED BY THE SECRETARY OF AGRICULTURE , BAERSON, Scott, R. , PAN, Zhiqiang , RIMANDO, Agnes, M. , DAYAN, Franck, E. , COOK, Daniel
发明人： BAERSON, Scott, R. , PAN, Zhiqiang , RIMANDO, Agnes, M. , DAYAN, Franck, E. , COOK, Daniel
IPC分类号： C12N15/52 , C12N9/00 , C12N15/63 , A01H5/00
CPC分类号： A61K31/713 , C12N9/00 , C12N15/8242 , C12N15/825 , C12N15/8257 , C12N15/8279
摘要： Sorghum is considered to be an allelopathic crop species, producing phytotoxins such as the lipid benzoquinone sorgoleone (2-hydroxy-5-methoxy-3-[(Z,Z)-8',l l ', 14'- pentadecatrieneJ-^-benzoquinone) which likely accounts for much of its allelopathic properties. Prior investigations into the biosynthesis of sorgoleone have suggested the participation of one or more alkylresorcinol synthases (ARS), which are type III polyketide synthases (PKS) that produce 5-alkylresorcinols using medium to long-chain fatty acyl-CoA starter units via iterative condensations with malonyl-CoA. Current evidence suggests that sorgoleone biosynthesis occurs exclusively in root hair cells, involving the synthesis of a 5- pentadecatrienyl resorcinol intermediate derived from an unusual 16:3 fatty acyl-CoA starter unit. To characterize the enzymes responsible for the biosynthesis of this alkylresorcinol intermediate, a previously-described expressed sequence tag (EST) database prepared from isolated root hairs was first mined for all PKS-I ike sequences. Quantitative real-time RT- PCR analyses revealed that two of these sequences were preferentially expressed in root hairs, and recombinant enzyme studies demonstrated that both sequences (designated ARSl and ARS2) encode ARS enzymes capable of accepting a variety of fatty acyi-CoA starter units. Furthermore, RNA interference (RNAi) experiments directed against ARSl and ARS2 resulted in the generation of multiple independent transformant events exhibiting dramatically reduced sorgoleone levels. Thus, both ARSl and ARS2 are likely to participate in the biosynthesis of sorgoleone inpJαntα. The sequences oϊARSl and ARS2 were also used to identify several rice genes encoding ARSs, which are likely involved in the production of defense-related alkyiresorcinols.
摘要翻译：高粱被认为是一种化感作物，产生植物毒素，如脂质苯醌山梨酮（2-羟基-5-甲氧基-3 - [（Z，Z）-8'，11'，14'-十五碳三烯基苯醌），这可能占其大部分的抗病性。对sorgoleone的生物合成的先前研究表明，一种或多种烷基间苯二酚合酶（ARS）的参与，其是使用中等至长链脂肪酰辅酶A起始单元通过迭代缩合产生5-烷基间苯二酚的III型聚酮化合物合酶（PKS）与丙二酰辅酶A。目前的证据表明，sorgoleone生物合成完全发生在根毛细胞中，涉及合成来自不寻常的16：3脂肪酰辅酶A启动子单元的5-十五碳烯基间苯二酚中间体。为了表征负责该烷基间苯二酚中间体的生物合成的酶，首先从所有PKS-I序列开始从分离的根毛制备的先前描述的表达序列标签（EST）数据库。定量实时RT-PCR分析显示，其中两个序列优先在根毛中表达，重组酶研究表明两个序列（命名为ARS1和ARS2）编码能够接受各种脂肪酰辅酶A起始单位的ARS酶。此外，针对ARS1和ARS2的RNA干扰（RNAi）实验导致产生显着降低的sorgoleone水平的多个独立转化体事件。因此，ARS1和ARS2两者都可能参与了山药中的sorgoleone的生物合成。序列o ARS1和ARS2也用于鉴定编码ARS的几个水稻基因，这些基因可能参与防御相关的烷基苯酚的生产。

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