Production of functional recombinant tyrosine hydroxylase by the BPV-1 expression plasmids in the cell cultures.
Bovine papilloma virus type-1 (BPV-1)-based expression plasmids TkBPVTH and CGalBPVTH encoding the rat tyrosine hydroxylase (TH) enzyme have been designed for the development of gene therapy for experimental Parkinson's disease. The aim of the present work was to examine the transfection of BPVTH plasmids to express a dopaminergic transgene in the monkey CV1-P fibroblast, rat C6 glioma and human NHA astrocyte cell cultures. The biological function of the transgene was estimated by analyzing the production of recombinant TH mRNA and protein, and the synthesis of L-dopa and dopamine. The highest transfection efficiency was obtained using TkBPVTH plasmids (5 microg). Furthermore, the expression of TkBPVTH plasmids was associated with significant synthesis of TH enzyme and L-dopa in the C6 and NHA cell cultures.
Department of Pharmacology and Toxicology, University of Kuopio, POB 1627, FIN-70211 Kuopio, Finland.
Pasi
P
Lampela
Marko
M
Huotari
Anu
A
Harjula
Lasse
L
Oinonen
Mart
M
Ustav
Seppo
S
Ylä-Herttuala
Pekka T
PT
Männistö
Atso
A
Raasmaja
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000502:physiology; D052688:Bovine papillomavirus 1 / Q000235:genetics*; D002478:Cells, Cultured; D016543:Central Nervous System Neoplasms / Q000235:genetics / Q000473:pathology; D004298:Dopamine / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D005910:Glioma / Q000235:genetics / Q000473:pathology; D006801:Humans; D007980:Levodopa / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000379:methods; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics* / Q000378:metabolism*
D012333:RNA, Messenger; D011994:Recombinant Proteins; D007980:Levodopa; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine