Sodium butyrate stimulates the synthesis of firefly luciferase in transfected CHO cells but levels of BiP chaperone are unaffected.
A stably transfected CHO cell line (LUCLEAD) was used where the coding region of native Firefly luciferase was linked to the 3'-UTR of the bovine growth hormone, and the 5'-nucleotides coding for the albumin signal peptide were linked to the N-terminal end of the luciferase coding region. Incubation of cells with 1 or 2 mM sodium butyrate (SB) for 72 h had no effect on cell growth since cultures reached confluency at the same time as control cells. Although cell cultures incubated with SB at a concentration of 4 mM were only about 60% confluent the luciferase content was about 5-fold higher than that in control cells. Cells incubated with either 1 or 2 mM SB showed intermediate levels of luciferase content. The amount of the chaperone BiP in the cells was not affected by incubation with SB. The results indicate that SB can be used to effectively promote synthesis of recombinant luciferase.
Department of Biochemistry and Molecular Biology, University of Bergen, Arstadveien 19, NO-5009 Bergen, Norway.
Ingeborg
I
Winge
Ian F
IF
Pryme
D000818:Animals; D002087:Butyrates / Q000494:pharmacology*; D016466:CHO Cells / Q000187:drug effects* / Q000201:enzymology*; D002352:Carrier Proteins / Q000096:biosynthesis / Q000187:drug effects* / Q000235:genetics; D002449:Cell Aggregation / Q000187:drug effects / Q000502:physiology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D001517:Coleoptera; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D008156:Luciferases / Q000096:biosynthesis / Q000187:drug effects* / Q000235:genetics; D018832:Molecular Chaperones / Q000096:biosynthesis / Q000187:drug effects* / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000187:drug effects* / Q000235:genetics; D013347:Subcellular Fractions
D002087:Butyrates; D002352:Carrier Proteins; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011993:Recombinant Fusion Proteins; D008156:Luciferases