An integrated strategy for efficient vector construction and multi-gene expression in Plasmodium falciparum.

ABSTRACT:

Background

The construction of plasmid vectors for transgene expression in the malaria parasite, Plasmodium falciparum, presents major technical hurdles. Traditional molecular cloning by restriction and ligation often yields deletions and re-arrangements when assembling low-complexity (A + T)-rich parasite DNA. Furthermore, the use of large 5'- and 3'- untranslated regions of DNA sequence (UTRs) to drive transgene transcription limits the number of expression cassettes that can be incorporated into plasmid vectors.

Methods

To address these challenges, two high fidelity cloning strategies, namely yeast homologous recombination and the Gibson assembly method, were evaluated for constructing P. falciparum vectors. Additionally, some general rules for reliably using the viral 2A

SUBMITTER: Wagner JC 

PROVIDER: S-EPMC3842810 | biostudies-literature | 2013 Oct

REPOSITORIES: biostudies-literature