CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion.

ABSTRACT: Here, we present chimera assembly by plasmid recovery and restriction enzyme site insertion (CAPRRESI). CAPRRESI benefits from many strengths of the original plasmid recovery method and introduces restriction enzyme digestion to ease DNA ligation reactions (required for chimera assembly). For this protocol, users clone wildtype genes into the same plasmid (pUC18 or pUC19). After the in silico selection of amino acid sequence regions where chimeras should be assembled, users obtain all the synonym DNA sequences that encode them. Ad hoc Perl scripts enable users to determine all synonym DNA sequences. After this step, another Perl script searches for restriction enzyme sites on all synonym DNA sequences. This in silico analysis is also performed using the ampicillin resistance gene (ampR) fo

SUBMITTER: Santillan O 

PROVIDER: S-EPMC5608506 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature