Book chapter
Chapter 17 - Analysis of Picornavirus Internal Ribosome Entry Site Function in Vivo
This chapter describes the use of an efficient transient expression system within mammalian cells for the analysis of the mechanism of picornavirus Ribonucleic acid (RNA) translation. Picornavirus RNAs contain, within their 5' noncoding regions, elements of about 450 bases that direct a cap-independent mechanism of translation.
This type of element is termed as Internal Ribosome Entry Site (IRES). The activity of these elements is usually assessed by determining their ability to direct translation of a second Open Reading Frame (ORF) within a synthetic dicistronic messenger RNA (mRNA). Such assays can be performed in vitro with the use of RNA transcripts to program rabbit reticulocyte lysate (RRL) translation systems.
However, it is often desirable to assay the function of these elements within intact cells. An efficient transient expression system within cells offers a convenient alternative to the cell-free systems. The chapter discusses in detail about the cap-dependent and cap-independent translation. The chapter also explains concepts related to the characteristics of: picornavirus IRES elements, and vaccinia/T7 RNA polymerase expression system.
The chapter analyzes the function of picornavirus IRES in vivo, elaborating upon the difference between cap-dependent and IRES-directed translation..
Language: | English |
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Year: | 1997 |
Pages: | 323-340 |
ISBN: | 0080537049 , 0123911915 , 0125875452 , 1281119326 , 9780080537047 , 9780123911919 , 9780125875455 and 9781281119322 |
Types: | Book chapter |
DOI: | 10.1016/B978-012587545-5/50018-5 |