Robert P. Bennett

ORFDB: an information resource linking scientific content to a high‐quality Open Reading Frame (ORF) collection

The ORFDB (http://orf.invitrogen.com/) represents an ongoing effort at Invitrogen Corporation to integrate relevant scientific data with an evolving collection of human and mouse Open Reading Frame (ORF) clones (Ultimate ORF Clones). The ORFDB serves as a central data warehouse enabling researchers to search the ORF collection through its web portal ORFBrowser, allowing researchers to find the Ultimate ORF clones by blast, keyword, GenBank accession, gene symbol, clone ID, Unigene ID, LocusLink ID or through functional relationships by browsing the collection via the Gene Ontology (GO) Browser. As of October 2003, the ORFDB contains 6200 human and 2870 mouse Ultimate ORF clones. All Ultimate ORF clones have been fully sequenced with high quality, and are matched to public reference protein sequences. In addition, the cloned ORFs have been extensively annotated across six categories: Gene, ORF, Clone Format, Protein, SNP and Genomic links, with the information assembled in a format termed the ORFCard. The ORFCard represents an information repository that documents the sequence quality, alignment with respect to public protein sequences, and the latest publicly available information associated with each human and mouse gene represented in the collection.

Erratum: “Protein delivery using VP22”

On p. 20 of the January 2002 issue, an author was omitted from the correspondence entitled “Protein delivery using VP22” by Robert P. Bennett and Brian Dalby. An additional author, Pamela M. Guy, also contributed to this work. The correct authors for the correspondence are Robert P. Bennett, Brian Dalby, and Pamela M.

10 – PROTEIN EXPRESSION IN MAMMALIAN CELLS USING SINDBIS VIRUS

This chapter focuses on the use of the Sindbis (SIN) virus for heterologous protein expression in mammalian cells. This virus, along with Semliki Forest virus (SFV), has a simple replication pathway and a broad host range that allows very high levels of heterologous protein expression in a variety of eukaryotic cell types. The chapter presents an introduction to the virus and genome structure. The life cycle and host range are also mentioned. Further, the applications of the SIN virus vector system are discussed. It is used for cell biology, immunology, virology, etc. The chapter discusses that SIN virus expression would be useful for a wider variety of applications if the time of expression could be lengthened and if the cytopathic effects could be limited. The SIN virus has the potential to be very useful as an expression tool in vivo . SIN has a very broad host range because it recognizes the high-affinity laminin receptor. Finally, the chapter concludes the discussion with the fact that SIN virus is growing in popularity for heterologous protein expression in tissue-culture cell lines and in vivo . New advancements such as cell-specific targeting and the use of DNA promoters should further the use of SIN as a expression tool in vivo and increase the use of the SIN system as a candidate for gene-therapy protocols.