Electroporation of rat pituitary (GH) cell lines: optimal parameters and effects on endogenous hormone production.
An efficient electroporation procedure was established for the genetic transformation of two clonal strains of hormone producing rat pituitary cells (GH12C1 and GH3). We used the bacterial chloramphenicol acetyltransferase (CAT) gene as reporter gene to determine optimal conditions for electroporation. The conditions found to be optimal, measured as expression of the highest CAT activity, were 240-300 V and a DNA concentration of 30-60 micrograms/ml in sucrose buffer. Cell viability was then about 50 per cent. Maximum CAT activity was seen 24 hours after electroporation. The electroporation procedure, in the presence or absence of DNA, caused a transient decrease in endogenous growth hormone (GH) and prolactin (PRL) production.
Paulssen
R H
RH
Institute of Medical Biochemistry, University of Oslo, Norway.
Paulssen
E J
EJ
Aleström
P
P
Gautvik
K M
KM
D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D004558:Electric Stimulation; D013006:Growth Hormone / Q000096:biosynthesis*; D010911:Pituitary Neoplasms; D010957:Plasmids; D011388:Prolactin / Q000096:biosynthesis*; D011401:Promoter Regions, Genetic; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism / Q000502:physiology
D011994:Recombinant Proteins; D011388:Prolactin; D013006:Growth Hormone; D015500:Chloramphenicol O-Acetyltransferase