Production of combinatorial libraries of fused genes by sequential transposition reactions - art. no. e119

Naumann TA; Goryshin IY; Reznikoff WS

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Production of combinatorial libraries of fused genes by sequential transposition reactions - art. no. e119

机译：通过顺序转座反应产生融合基因的组合文库-艺术。没有。 e119

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The use of in vivo and in vitro transposition reactions to perform non-combinatorial manipulation of DNAs in molecular biology is widespread. In this work we describe a technique that utilizes two sequential, directed transposition reactions in order to carry out combinatorial DNA manipulations. The methodology relies on the use of two different mutant Tn5 transposase proteins that have different transposon end recognition specificities. We demonstrate that the technique can be used to create large libraries of random fusions between two genes. These transpositional fusions are defined by insertion of a 32 bp linker sequence. We applied the technique to a model system, chloramphenicol acetyl transferase, to create functional fusions from N- and C-terminally truncated, non-functional genes. Comparative structural analysis suggests that both sides of the linker are inserted into disordered regions in functional proteins.

机译：体内和体外转座反应在分子生物学中广泛用于DNA的非组合操作。在这项工作中，我们描述了一种技术，该技术利用两个顺序的定向转座反应来进行组合DNA操纵。该方法依赖于使用具有不同转座子末端识别特异性的两种不同的突变Tn5转座酶蛋白。我们证明该技术可用于创建两个基因之间随机融合的大型库。这些转座融合体是通过插入32 bp的接头序列来定义的。我们将该技术应用于模型系统氯霉素乙酰转移酶，以从N和C末端截短的非功能性基因创建功能性融合体。比较结构分析表明，接头的两侧均插入功能蛋白的无序区域。

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《Nucleic Acids Research》 |2002年第21期|共1页
作者
Naumann TA; Goryshin IY; Reznikoff WS;
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正文语种 eng
中图分类细胞形态学;生物化学;
关键词
In-vitro; Tn5 transposition; Mutagenesis; Integration; Efficient; Protein;

机译：体外Tn5转座诱变整合高效蛋白;

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1. Production of combinatorial libraries of fused genes by sequential transposition reactions - art. no. e119 [J] . Naumann TA, Goryshin IY, Reznikoff WS Nucleic Acids Research . 2002,第21期

机译：通过顺序转座反应产生融合基因的组合文库-艺术。没有。 e119
2. Simple and straightforward construction of a mouse gene targeting vector using in vitro transposition reactions - art. no. e52 [J] . Aoyama M, Agari K, Sun-Wada GH, Nucleic Acids Research . 2005,第5期

机译：使用体外转座反应简单，直接构建小鼠基因靶向载体-艺术。没有。 e52
3. Designing gene libraries from protein profiles for combinatorial protein experiments - art. no. e120 [J] . Wang W, Saven JG Nucleic Acids Research . 2002,第21期

机译：从蛋白质谱设计基因库以进行组合蛋白质实验-艺术。没有。 e120
4. Combinatorial Approaches for New Catalyst Discovery: The First Homogeneous Catalysts Derived from Combinatorial Technologies for a Variety of Cross-Coupling Reactions [C] . George Y. Li Conference on the Catalysis of Organic Reactions . 2005

机译：新型催化剂的组合方法：第一种衍生自组合技术的均相催化剂，用于各种交叉偶联反应
5. Optimization of Protein Bioconjugation Reactions Using Combinatorial Peptide Libraries. [D] . Witus, Leah Suzanne. 2012

机译：使用组合肽库优化蛋白质生物偶联反应。
6. Production of combinatorial libraries of fused genes by sequential transposition reactions [O] . Todd A. Naumann, Igor Y. Goryshin, William S. Reznikoff 2002

机译：通过顺序转座反应产生融合基因的组合文库
7. Production of combinatorial libraries of fused genes by sequential transposition reactions [O] . Naumann, Todd A., Goryshin, Igor Y., Reznikoff, William S. 2002

机译：通过顺序转座反应产生融合基因的组合文库

Production of combinatorial libraries of fused genes by sequential transposition reactions - art. no. e119

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