A new method to customize protein expression vectors for fast, efficient and background free parallel cloning.

ABSTRACT:

Background

Expression and purification of correctly folded proteins typically require screening of different parameters such as protein variants, solubility enhancing tags or expression hosts. Parallel vector series that cover all variations are available, but not without compromise. We have established a fast, efficient and absolutely background free cloning approach that can be applied to any selected vector.

Results

Here we describe a method to tailor selected expression vectors for parallel Sequence and Ligation Independent Cloning. SLIC cloning enables precise and sequence independent engineering and is based on joining vector and insert with 15-25 bp homologies on both DNA ends by homologous recombination. We modified expression vectors based on pET, pFastBac and pTT b

SUBMITTER: Scholz J 

PROVIDER: S-EPMC3598636 | biostudies-literature | 2013 Feb

REPOSITORIES: biostudies-literature