N-linked protein glycosylation is the most common post-translational modification made to secretory and membrane proteins of eukaryotes and serves to expand the diversity of the proteome for these organisms. The process of N-linked glycosylation was originally considered to be restricted to eukaryotes, however recent evidence confirms that a similar, albeit less complex, process is performed in certain prokaryotes. Like its eukaryotic counterpart, prokaryotic N-glycan addition occurs via the attachment of glycan structures to target proteins by an oligosaccharyltransferease (OST) at an Asn-Xaa-Ser/Thr consensus sequence.  Also similar is the fact that both prokaryotic and eukaryotic OST systems attach glycans in functionally equivalent extracytoplasmic compartments, namely the endoplasmic reticulum (ER) and periplasm, respectively. There are also many notable differences between the two systems that serve to limit the utility of the bacterial glycosylation machinery for biotechnology applications in which human-like glycosylation patterns are desired. To address this and other limitations, we have engineered a collection of genetic tools for manipulation of the bacterial N-linked glycosylation machinery. First, using synthetic E. coli strains that can perform recombinant N-linked protein glycosylation, we have engineered two protein display strategies for directed evolution of the glycosylation pathway such as the OST or the carbohydrate biosynthesis enzymes. Second, we have developed a novel in vivo two-hybrid-like assay for analyzing protein-carbohydrate interactions directly in the periplasm of living cells. These approaches represent the first crucial steps towards engineering bacterial cells with human-like bacterial glycosylation systems and are expected to open the door to glycoengineering in Gram-negative bacteria.