HomeRun Vector Assembly System: a flexible and standardized cloning system for assembly of multi-modular DNA constructs.

ABSTRACT: Advances in molecular and synthetic biology call for efficient assembly of multi-modular DNA constructs. We hereby present a novel modular cloning method that obviates the need for restriction endonucleases and significantly improves the efficiency in the design and construction of complex DNA molecules by standardizing all DNA elements and cloning reactions. Our system, named HomeRun Vector Assembly System (HVAS), employs a three-tiered vector series that utilizes both multisite gateway cloning and homing endonucleases, with the former building individual functional modules and the latter linking modules into the final construct. As a proof-of-principle, we first built a two-module construct that supported doxycycline-induced expression of green fluorescent protein (GFP). Further, with a

SUBMITTER: Li MV 

PROVIDER: S-EPMC4069157 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature