公开(公告)号：US5798240A

公开(公告)日：1998-08-25

申请号：US584226

申请日：1996-01-11

申请人： Susan A. Martinis ,  Mandana Sassanfar ,  Sunghoon Kim ,  Sang Ho Lee ,  Paul R. Schimmel

发明人： Susan A. Martinis ,  Mandana Sassanfar ,  Sunghoon Kim ,  Sang Ho Lee ,  Paul R. Schimmel

IPC分类号： A61K38/00 ,  C12N9/00 ,  C07H21/04 ,  C12N1/20 ,  C12N15/00

CPC分类号： C12N9/93 ,  A61K38/00 ,  Y10S435/863

摘要： Isolated and/or recombinant nucleic acids encoding mycobacterial methionyl-tRNA synthetase have been characterized. Recombinant DNA constructs and vectors having a sequence which encodes mycobacterial methionyl-tRNA synthetase have been made, and can be used for the construction of tester strains as well as for the production of isolated and/or recombinant methionyl-tRNA synthetases. These enzymes or portions thereof are useful in the biochemical separation of methionine and quantification of methionine or ATP, and for producing antibodies useful in the purification and study of the enzyme, for example. Host cells and methods useful for producing recombinant mycobacterial methionyl-tRNA synthetases are described, as are tester strains, which are cells engineered to rely on the function of the tRNA synthetase encoded by an introduced cloned gene. Tester strains can be used to identify inhibitors of the essential tRNA synthetase enzyme encoded by the introduced cloned gene, and thus provide a means to assess the antimicrobial effect and specificity of the inhibitor without employing slow-growing, pathogenic strains of mycobacteria, such as Mycobacterium tuberculosis.

摘要翻译： 已经表征了编码分枝杆菌甲硫氨酰-tRNA合成酶的分离和/或重组核酸。 已经制备了具有编码分枝杆菌甲硫氨酰-tRNA合成酶的序列的重组DNA构建体和载体，并且可用于构建测试菌株以及用于分离和/或重组甲硫氨酰-tRNA合成酶的产生。 这些酶或其部分可用于甲硫氨酸的生物化学分离和甲硫氨酸或ATP的定量，以及用于产生用于纯化和研究酶的抗体。 描述了可用于产生重组分枝杆菌甲硫氨酰-tRNA合成酶的宿主细胞和方法，试验菌株是被依赖于引入的克隆基因编码的tRNA合成酶的功能而被工程化的细胞。 测试菌株可以用于鉴定由引入的克隆基因编码的必需tRNA合成酶的抑制剂，并且因此提供了评估抑制剂的抗微生物效果和特异性的方法，而不使用分枝杆菌的缓慢生长的致病菌株，例如分枝杆菌 结核。

公开(公告)号：US07785827B2

公开(公告)日：2010-08-31

申请号：US10251648

申请日：2002-09-20

申请人： Susan A. Martinis ,  James M. Briggs ,  Richard S. Mursinna ,  Keun Woo Lee ,  Tommie L. Lincecum ,  Amy M. Williams ,  Yuxin Zhai

发明人： Susan A. Martinis ,  James M. Briggs ,  Richard S. Mursinna ,  Keun Woo Lee ,  Tommie L. Lincecum ,  Amy M. Williams ,  Yuxin Zhai

IPC分类号： C07K14/47 ,  C07H21/04 ,  C12P21/06

CPC分类号： C12N9/93

摘要： A method and composition for tRNA synthetases that activate and aminoacylate nonstandard and noncognate amino acids to tRNA adaptor molecules is described that can be used to generate custom designed protein products for uses in medicinal, therapeutic, diagnostic, biotechnology, engineering, and spectroscopy applications. Some tRNA synthetases naturally misactivate and misaminoacylate noncognate amino acids. Many of these tRNA synthetases, including but not limited to leucyl-, isoleucyl-, and valyl-tRNA synthetases, have evolved proofreading and editing mechanisms to correct these mistakes. Inactivation of the enzyme's editing activity allows and facilitates production and accumulation of tRNAs that are misaminoacylated with nonstandard and noncognate amino acids. These misaminoacylated tRNAs can be used to introduce novel amino acids into proteins.

摘要翻译： 描述了用于将非标准和非认可氨基酸激活和氨基化至tRNA衔接子分子的tRNA合成酶的方法和组合物，其可用于产生用于药物，治疗，诊断，生物技术，工程和光谱学应用的定制设计的蛋白质产物。 一些tRNA合成酶天然失活和非氨基酸化非识别氨基酸。 许多这些tRNA合成酶，包括但不限于亮氨酰，异亮氨酰和缬氨酰-tRNA合成酶，已经发展了校正和编辑机制，以纠正这些错误。 酶的编辑活性的失活允许并促进用非标准和非认知氨基酸错氨基酰化的tRNA的产生和积累。 这些错氨基酰化的tRNA可用于将新的氨基酸引入蛋白质。

公开(公告)号：US5656470A

公开(公告)日：1997-08-12

申请号：US305172

申请日：1994-09-13

申请人： Susan A. Martinis ,  Jiansu Zhang ,  Paul R. Schimmel

发明人： Susan A. Martinis ,  Jiansu Zhang ,  Paul R. Schimmel

IPC分类号： C12N9/00 ,  C07H21/04 ,  C12N1/20 ,  C12N15/00

CPC分类号： C12N9/93

摘要： Isolated and/or recombinant nucleic acids encoding mycobacterial seryl-tRNA synthetase have been characterized. Recombinant DNA constructs and vectors having a sequence which encodes mycobacterial seryl-tRNA synthetase have been made, and can be used for the construction of tester strains as well as for the production of isolated and/or recombinant seryl-tRNA synthetases. These enzymes or portions thereof are useful in the biochemical separation of serine and quantification of serine or ATP, and for producing antibodies useful in the purification and study of the enzyme, for example. Host cells and methods useful for producing recombinant mycobacterial seryl-tRNA synthetases are described, as are tester strains, which are cells engineered to rely on the function of the tRNA synthetase encoded by an introduced cloned gene. Tester strains can be used to identify inhibitors of the essential tRNA synthetase enzyme encoded by the introduced cloned gene, and thus provide a means to assess the antimicrobial effect and specificity of the inhibitor without employing slow-growing, pathogenic strains of mycobacteria, such as Mycobacterium tuberculosis.

摘要翻译： 已经表征了编码分枝杆菌丝氨酰-tRNA合成酶的分离和/或重组核酸。 已经制备了具有编码分枝杆菌丝氨酰-tRNA合成酶的序列的重组DNA构建体和载体，并且可用于构建测试菌株以及分离和/或重组丝氨酰-tRNA合成酶的生产。 这些酶或其部分可用于例如丝氨酸的生物化学分离和丝氨酸或ATP的定量，以及用于产生用于纯化和研究酶的抗体。 描述了可用于产生重组分枝杆菌丝氨酰-tRNA合成酶的宿主细胞和方法，试验菌株是被依赖于引入的克隆基因编码的tRNA合成酶的功能而被工程化的细胞。 测试菌株可以用于鉴定由引入的克隆基因编码的必需tRNA合成酶的抑制剂，并且因此提供了评估抑制剂的抗微生物效果和特异性的方法，而不使用分枝杆菌的缓慢生长的致病菌株，例如分枝杆菌 结核。