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    • 6. 发明申请
    • GENETICALLY ENGINEERED MICROBES AND METHODS FOR PRODUCING CITRAMALATE
    • 遗传工程微生物和生产CITRAMALATE的方法
    • WO2017165397A1
    • 2017-09-28
    • PCT/US2017/023380
    • 2017-03-21
    • UNIVERSITY OF GEORGIA RESEARCH FOUNDATION, INC.WU, XianghaoEITEMAN, Mark
    • WU, XianghaoEITEMAN, Mark
    • C12N15/70C12N9/10C12P7/46
    • C12P7/46C12N9/10C12N9/1025C12N9/1029C12N15/70C12Y203/01182C12Y203/03001
    • Provided herein is a genetically engineered microbe which accumulates citramalate. In one embodiment, the microbe includes an exogenous polynucleotide encoding a citramalate synthase which catalyzes the condensation of acetyl CoA and pyruvic acid. Optionally, the microbe also includes a second exogenous polynucleotide encoding a citrate synthase which catalyzes the condensation of acetyl CoA and oxaloacetate, and the citrate synthase activity in the microbe is reduced compared to a control microbe. In one embodiment, the citrate synthase includes at least one amino acid substitution in the acetyl-CoA binding pocket, the mobile loop, the NADH binding site, and the oxaloacetate binding site, or a combination thereof. Also provided herein are methods for using the genetically engineered microbe, including a method is for producing citramalate. The method can further include isolating the citramalate.
    • 本文提供了一种积聚柠苹果酸酯的基因工程微生物。 在一个实施方案中,微生物包括编码柠檬酸苹果酸合酶的外源多核苷酸,其催化乙酰CoA和丙酮酸的缩合。 任选地,微生物还包括编码柠檬酸合酶的第二外源多核苷酸,其催化乙酰CoA和草酰乙酸的缩合,并且微生物中的柠檬酸合酶活性与对照微生物相比降低。 在一个实施方案中,柠檬酸合酶在乙酰-CoA结合袋,移动环,NADH结合位点和草酰乙酸结合位点或其组合中包含至少一个氨基酸取代。 本文还提供了使用基因工程微生物的方法,包括生产柠苹酸的方法。 该方法可以进一步包括分离柠檬酸苹果酸酯。
    • 8. 发明申请
    • PICHINDE VIRUS REVERSE GENETICS SYSTEM AND METHODS OF USE
    • PICHINDE VIRUS逆向遗传系统及其使用方法
    • WO2016048949A1
    • 2016-03-31
    • PCT/US2015/051337
    • 2015-09-22
    • REGENTS OF THE UNIVERSITY OF MINNESOTALY, HinhLIANG, Yuying
    • LY, HinhLIANG, Yuying
    • C12N7/00C12N15/86A61K39/00
    • C12N7/00A61K2039/5256A61K2039/70C12N15/86C12N2760/10043C12N2760/16134
    • Provided herein are genetically engineered Pichinde viruses that include three ambisense genomic segments. The first genomic segment includes a coding region encoding a Z protein and a coding region encoding a L RdRp protein. The second genomic segment includes a coding region encoding a nucleoprotein (NP) and the third genomic segment includes a coding region encoding a glycoprotein. Each of the second and third genomic segments optionally include an additional coding region that may encode an antigen or a detectable marker. Also provided herein is a reverse genetics system for making a genetically engineered Pichinde virus, and a collection of vectors that can be used to produce a genetically engineered Pichinde virus. Further provided are methods for using a reverse genetics system, and methods for producing an immune response in a subject.
    • 本文提供了包括三个ambisense基因组片段的基因工程的Pichinde病毒。 第一基因组片段包括编码Z蛋白的编码区和编码L RdRp蛋白的编码区。 第二基因组片段包括编码核蛋白(NP)的编码区,第三基因组片段包括编码糖蛋白的编码区。 第二和第三基因组片段中的每一个任选地包括可以编码抗原或可检测标记的另外的编码区。 本文还提供了用于制备基因工程的Pichinde病毒的反向遗传学系统,以及可用于产生遗传工程化的Pichinde病毒的载体的集合。 还提供了使用反向遗传学系统的方法和用于在受试者中产生免疫应答的方法。