Transient expression in mammalian cells of the bacterial reporter gene encoding mercuric reductase: effects of various regulatory elements.
The effect of several transcriptional regulatory elements on gene expression in mammalian cells was investigated. As a reporter gene we have used the bacterial gene merA coding for the enzyme mercuric reductase. Several plasmids were constructed with different promoter/enhancer sequences (pSV/E, pSV/L, pMT, pRSV or pAd) at the 5' end and different splicing (small intron of the T antigen of SV40 or the second intron of the rabbit beta-globin gene) and/or polyadenylation signals (AEn, ALn or AR beta Gn) at the 3' end of the merA gene. Expression was measured in five different mammalian cell lines. In COS cells the highest level of expression is obtained with pSV/L and the lowest level with pSV/E. In HeLa, CV-1, Ltk-, and CHO cells merA expression is relatively high, under control of pRSV and pMT and relatively low under control of pSV/L and pAd. The introns studied have a negative effect on the expression of merA. The presence of a polyadenylation signal downstream from the gene is essential for its expression. The three different polyadenylation signals studied give a similar stimulatory effect on the level of expression of the merA gene.
Veldhuisen
G
G
TNO Medical Biological Laboratory, Rijswijk, The Netherlands.
Van Dijk
M
M
Meijer
J
J
Enger-Valk
B E
BE
Pouwels
P H
PH
D000818:Animals; D002478:Cells, Cultured; D004742:Enhancer Elements, Genetic; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D005819:Genetic Markers; D010088:Oxidoreductases / Q000235:genetics*; D010430:Pentosyltransferases / Q000032:analysis; D010957:Plasmids; D011061:Poly A / Q000235:genetics; D011401:Promoter Regions, Genetic; D014162:Transfection
D005819:Genetic Markers; D011061:Poly A; D010088:Oxidoreductases; C021551:mercuric reductase; D010430:Pentosyltransferases; C024827:xanthine phosphoribosyltransferase