Automated seamless DNA co-transformation cloning with direct expression vectors applying positive or negative insert selection.

ABSTRACT:

Background

Molecular DNA cloning is crucial to many experiments and with the trend to higher throughput of modern approaches automated techniques are urgently required. We have established an automated, fast and flexible low-cost expression cloning approach requiring only vector and insert amplification by PCR and co-transformation of the products.

Results

Our vectors apply positive selection for the insert or negative selection against empty vector molecules and drive strong expression of target proteins in E.coli cells. Variable tags are available both in N-terminal or C-terminal position. A newly developed beta-lactamase (DeltaW290) selection cassette contains a segment inside the beta-lactamase open reading frame encoding a stretch of hydrophilic amino acids that result

SUBMITTER: Olieric N 

PROVIDER: S-EPMC2924254 | biostudies-literature | 2010 Aug

REPOSITORIES: biostudies-literature