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[18] Applications of oriP plasmids and their mode of replication

1999; Academic Press; Linguagem: Inglês

10.1016/s0076-6879(99)06020-6

1557-7988

David Mackey, Bill Sugden,

CRISPR and Genetic Engineering

This chapter discusses various applications of oriP plasmids, including the generation of cell lines stably or inducibly expressing a cloned gene. The chapter discusses the study of the regulation of transcription from a cloned promoter and the study of mutagenesis of a particular target sequence. It discusses the isolation of plasmids containing desired cDNAs for which positive elections are available. Replication of oriP requires a single viral trans-acting factor, Epstein-Barr nuclear antigen 1 (EBNA1). In cells expressing EBNA1, plasmids containing oriP replicate during S phase and segregate to daughter cells efficiently. EBNA1 and oriP, therefore, provide a model system for the study of replication and segregation of DNAs in mammalian cells. To provide a framework for the informed application of oriP plasmids, the chapter describes the present understanding of their function.