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Recursive Directional Ligation Approach for Cloning Recombinant Spider Silks.

Methods Mol. Biol.. 2018; 
DinjaskiNina,HuangWenwen,KaplanDav
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Gene Synthesis … 3a, Note 1 ). The A and B block sequences are synthesized as the NheI/SpeI insets (GenScript, Piscataway, NJ, USA) in pUC57 (Kn) … 2. Annealed forward and reverse cloning cassette linker (1:1 v/v) (GenScript, Piscataway, NJ, USA). 3 … Get A Quote

摘要

Recent advances in genetic engineering have provided a route to produce various types of recombinant spider silks. Different cloning strategies have been applied to achieve this goal (e.g., concatemerization, step-by-step ligation, recursive directional ligation). Here we describe recursive directional ligation as an approach that allows for facile modularity and control over the size of the genetic cassettes. This approach is based on sequential ligation of genetic cassettes (monomers) where the junctions between them are formed without interrupting key gene sequences with additional base pairs.

关键词

Cloning,Recombinant DNA techniques,Recursive ligation,Spider