Downstream insertion of the adenovirus tripartite leader sequence enhances expression in universal eukaryotic vectors.
A series of universal eukaryotic gene expression vectors was constructed. All vectors contain a viral promoter and enhancer, a polylinker for insertion of the gene of interest and a polyadenylation sequence. To enhance translation, we inserted the tripartite leader sequence of an adenovirus downstream of the promoter. Using the chloramphenicol acetyl transferase (cat) gene as a marker, we show that the strength of various promoters/enhancers in different cell lines differed by two orders of magnitude. The presence of the tripartite leader increased the efficiency of gene expression up to 18-fold. The level of increase is promoter specific and is most likely influenced by additional sequences flanking the tripartite leader sequence.
Sheay
W
W
Dept. of Microbiology, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway 08854.
Nelson
S
S
Martinez
I
I
Chu
T H
TH
Bhatia
S
S
Dornburg
R
R
D016475:3T3 Cells; D000256:Adenoviridae / Q000235:genetics*; D000667:Ampicillin; D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003587:Cytomegalovirus / Q000235:genetics; D004261:DNA Replication; D004285:Dogs; D004351:Drug Resistance / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics; D012516:Osteosarcoma; D010957:Plasmids; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured
D021382:Protein Sorting Signals; D000667:Ampicillin; D015500:Chloramphenicol O-Acetyltransferase