Activity assays of nine heterogeneous promoters in neural and other cultured cells.
To express high levels of proteins encoded by transfected DNA constructs in a variety of cultured cells, including neuronal cells, the activities of nine different promoters were evaluated using Escherichia coli beta-galactosidase (beta-gal) (LacZ) as a reporter gene. These nine promoters were categorized into three distinct groups (high, intermediate, and low expresser), in terms of the levels of beta-gal expression. An expression vector containing the cytomegalovirus enhancer and the chick beta-actin promoter (high expresser) showed the highest levels of expression, followed by vectors containing the cytomegalovirus promoter/enhancer and the SV40 promoter/enhancer (intermediate expresser). The rest of the promoters (thymidine kinase, adenovirus, murine proliferative sarcoma virus, nerve growth factor receptor, Rous sarcoma and mouse mammary tumor virus, and beta-amyloid precursor protein) expressed low levels of beta-gal. These results were consistent for eight different cell types. A particularly attractive model is the stem cell, P19; cultures differentiating into progeny consisting predominantly of cholinergic neurons could be readily transfected with expression vectors using liposomes and expressed beta-gal without significant morphologic changes of the differentiated neurons. The systems should be useful for the study of promoters and various expressed proteins, including those involved in axonal transport.
Fukuchi
K
K
Department of Pathology, University of Washington, Seattle.
Hearn
M G
MG
Deeb
S S
SS
Smith
A C
AC
Dang
N
N
Miyazaki
J
J
Bothwell
M
M
Martin
G M
GM
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002522:Chlorocebus aethiops; D004742:Enhancer Elements, Genetic; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005069:Evaluation Studies as Topic; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D014162:Transfection; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000235:genetics
D014212:Tretinoin; D001616:beta-Galactosidase