Use of the Gateway System for Protein Expression in Multiple Hosts

The Gateway cloning method allows a gene to be cloned and subsequently transferred into any vector by in vitro site-specific recombination. It does not necessarily follow, however, that a gene can be cloned once and expressed in all the available host-vector combinations with uniformly satisfactory results. This is because different organisms have different mechanisms of translating mRNA into protein, and also because choices always have to be made when designing an expression construct, for example, the presence or absence of a stop codon. This unit reviews Gateway cloning, summarizes aspects of protein expression that limit the universality of the use of one clone in many vectors and hosts, and discusses how conflicts between the structure of a Gateway clone of a gene and the rules of protein expression can be minimized or resolved.The Gateway cloning method allows a gene to be cloned and subsequently transferred into any vector by in vitro site-specific.