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Titolo:
Efficient DNA transfection in neuronal and astrocytic cell lines
Autore:
Ghosh, C; Song, W; Lahiri, DK;
Indirizzi:
Indiana Univ, Sch Med, Dept Psychiat, Lab Mol Neurogenet,Inst Psychiat Res, Indianapolis, IN 46202 USA Indiana Univ Indianapolis IN USA 46202 at Res, Indianapolis, IN 46202 USA Harvard Univ, Childrens Hosp, Sch Med, Boston, MA 02115 USA Harvard Univ Boston MA USA 02115 rens Hosp, Sch Med, Boston, MA 02115 USA
Titolo Testata:
MOLECULAR BIOLOGY REPORTS
fascicolo: 2, volume: 27, anno: 2000,
pagine: 113 - 121
SICI:
0301-4851(200006)27:2<113:EDTINA>2.0.ZU;2-8
Fonte:
ISI
Lingua:
ENG
Soggetto:
AMYLOID PRECURSOR PROTEIN; MEDIATED GENE-TRANSFER; MAMMALIAN-CELLS; CALCIUM-PHOSPHATE; PROMOTER ACTIVITY; ELECTROPORATION; TRANSFORMATION; EXPRESSION;
Keywords:
beta-amyloid precursor protein; calcium phosphate; electroporation; neuroblastoma cells; PC12 cells; promoter; reporter gene;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
27
Recensione:
Indirizzi per estratti:
Indirizzo: Lahiri, DK Indiana Univ, Sch Med, Dept Psychiat, Lab Mol Neurogenet,Inst Psychiat Res, 791 Union Dr, Indianapolis, IN 46202 USA Indiana Univ 791 Union Dr Indianapolis IN USA 46202 N 46202 USA
Citazione:
C. Ghosh et al., "Efficient DNA transfection in neuronal and astrocytic cell lines", MOL BIOL RP, 27(2), 2000, pp. 113-121

Abstract

We have studied different parameters for efficient DNA transfection in various cell types and with different size of the promoter. Here we report that the optimum condition for DNA transfection by electroporation is 350 V/960 muF for PC12, 450V/960 muF C6 cells, and 250 V/500 muF for COS-1 cells. For the human neuroblastoma (SK-N-SH) cells the optimum condition for DNA transfection is by the calcium phosphate method. In promoter mapping studies,a serial deletion approach is commonly used. To optimize transfection we have selected three DNA constructs that varied in size from 4.5 to 12.4 kilobases (kb). We measured the promoter activity of these constructs under conditions of 'equal amount', 'equimolar', and 'equimolar plus carrier DNA to make it equal amount'. We recommend that for comparative purpose, transfection should be carried out under 'equimolar condition' without a need to adjust the total amount of DNA by carrier DNA. Taken together, our results suggest that efficient methods for DNA transfection are important to study gene regulation by devising better ways to deliver DNA into the mammalian cells.

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Documento generato il 04/06/20 alle ore 01:19:17