Importance of SV40 early 5′ distal sequences in directing heterologous gene expression

Abstract

The chloramphenicol acetyltransferase-encoding reporter gene ( cat) is used extensively in assessing the ability of transcriptional regulatory elements ( TRE) to direct gene expression in eukaryotic cells. Two commonly utilized plasmids contain the cat coding sequences under the transcriptional control of the Rous sarcoma virus LTR (pRSV cat) or simian virus 40 early (SV40E) promoter (pSV2 cat). In the present study, we have recloned the RSV-LTR and SV40E TRE into a pUC18 vector. Direct comparison of these TRE in different plasmid vectors, as well as reevaluation of their relative level of cat expression revealed: ( 1) a small but significant increases in SV40E-directed reporter gene expression was observed when the TRE was inserted into the pUC18 vector; and ( 2) a significant increase in SV40E-directed gene expression was realized by inclusion of the 69-bp 5′ of the sequences present in pSV2 cat. These distal sequences are required for maximal activity of the SV40 TRE in the cell lines tested.