Recursive Directional Ligation Approach for Cloning Recombinant Spider Silks

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1777)

Abstract

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.

Key words

Cloning Recombinant DNA techniques Recursive ligation Spider silk 

Notes

Acknowledgments

We thank the NIH (P41 EB002520), the AFOSR, and the NSF for support of this work.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Biomedical EngineeringTufts UniversityMedfordUSA

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