[Studies on the transcriptional regulatory elements of the hCG beta gene cluster using a choriocarcinoma cell line which expresses hCG beta eutopically].
We previously identified the promoter region for hCG beta gene with mouse adrenal cell line Y1. However, it was not know why this cell line, which does not express hCG beta eutopically, expresses hCG beta when the gene is transfected exogenously. Therefore, to confirm the results obtained in Y1 cell experiments, it was important to establish a system in which the activity of hCG beta transcriptional regulatory elements can be tested in cells which express hCG beta eutopically. Here we report success in establishing such a system, using constructs containing hCG beta upstream elements in front of the promotorless chloramphenicol acetyl transferase (CAT) gene and also carrying the neomycin resistance gene. These constructs were transfected stably into JAr choriocarcinoma cells which express hCG beta eutopically. Then the expression of CAT in each transfected cell line was examined. The results of this experiment confirmed the presence of basic promoter elements within 78bp of the transcriptional initiation site, which we had suggested previously as a result of Y1 experiments. The results also suggest the existence of additional transcriptional regulatory elements further upstream. It was also confirmed that gene 7 is an inactive gene.
Otani
T
T
Department of Obstetrics and Gynecology, Kobe University School of Medicine.
Boime
I
I
Mochizuki
M
M
D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002822:Choriocarcinoma / Q000235:genetics*; D006063:Chorionic Gonadotropin / Q000235:genetics*; D018997:Chorionic Gonadotropin, beta Subunit, Human; D002874:Chromosome Mapping; D005260:Female; D015870:Gene Expression; D005809:Genes, Regulator; D006801:Humans; D005810:Multigene Family; D010446:Peptide Fragments / Q000235:genetics*; D010957:Plasmids; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014162:Transfection; D014594:Uterine Neoplasms / Q000235:genetics*
D006063:Chorionic Gonadotropin; D018997:Chorionic Gonadotropin, beta Subunit, Human; D010446:Peptide Fragments; D015500:Chloramphenicol O-Acetyltransferase