Adenoviral Vector Construction II: Bacterial Systems

Publisher Summary This chapter describes the construction of adenoviral vectors for bacterial systems. It describes the recent development of novel technologies, which efficiently permit the rapid construction and generation of single or multiply deleted Ad in Escherichia coli. Extensive genetic and molecular analyses of adenovirus (Ad) have resulted in a detailed knowledge of the viral life cycle and the function of the majority of viral proteins, further stimulating the use and modifications of Ad vectors. Three methods, namely, homologous recombination, direct ligation and cosmid technology have evolved to clonally derive Ad constructs in Escherichia coli . The advent of these technologies has enabled the construction and generation of numerous modified Ad vectors carrying many different transgenes. Many applications of these methods can be envisioned, including the use of custom tailored Ad for the in vivo production of recombinant therapeutic proteins or the generation of custom-made libraries in Ad vectors to enable the high-throughput screening and applied functional analysis of many genes in the context of functional genomics projects.