Factors affecting polybrene-mediated transfection of cultured Aedes albopictus (mosquito) cells

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Abstract

The polycation 1,5-dimethyl-1,5-diazaundecamethylene polymethobromide (polybrene) is superior to calcium phosphate for the introduction of purified DNA into cultured Aedes albopictus (mosquito) cells. Adsorption of the polybrene-DNA complex to mosquito cells was essentially linear for 6 h. However, the rate of adsorption of DNA increased when the DNA-polybrene mixture was preincubated for several hours prior to addition to cells. A recombinant plasmid carrying an inducible chloramphenicol acetyltransferase gene under the control of a Drosophila heat shock protein (hsp) promoter was used to show that expression of transfected DNA was highest when cells were treated with a freshly prepared polybrene-DNA mixture. Optimal expression was observed in cells transfected with 4-13 μg of DNA per 106 cells; transfection with 24 μg of DNA resulted in reduced CAT expression. Variation in the polybrene-DNA ratio improved transfection with high levels of DNA. In mosquito cells, CAT expression was independent of DNA methylation.

Original languageEnglish (US)
Pages (from-to)535-542
Number of pages8
JournalExperimental Cell Research
Volume166
Issue number2
DOIs
StatePublished - Jan 1 1986

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Hexadimethrine Bromide
Aedes
Culicidae
Transfection
DNA
Adsorption
Drosophila Proteins
Chloramphenicol O-Acetyltransferase
DNA Methylation
Heat-Shock Proteins
Plasmids

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Factors affecting polybrene-mediated transfection of cultured Aedes albopictus (mosquito) cells. / Fallon, Ann Marie.

In: Experimental Cell Research, Vol. 166, No. 2, 01.01.1986, p. 535-542.

Research output: Contribution to journalArticle

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