Efficient transfection of mosquito cells is influenced by the temperature at which DNA-calcium phosphate coprecipitates are prepared.
Transient expression of a heat-shock protein-chloramphenicol acetyltransferase (hsp-CAT) recombinant plasmid was used to define the parameters that influence transfection of cultured mosquito cells using DNA-calcium phosphate coprecipitates. The efficiency of the calcium phosphate procedure was strongly influenced by the growth state of recipient cells, and by the temperature at which the coprecipitate was prepared. Under optimal conditions, which included formation of the DNA-calcium phosphate coprecipitate at 50 degrees C, transfection frequencies were up to tenfold higher than those obtained using the previously described polybrene procedure.
Kjer
K M
KM
Department of Entomology, University of Minnesota, St. Paul 55108.
Fallon
A M
AM
D000330:Aedes; D000818:Animals; D002130:Calcium Phosphates; D002460:Cell Line; D002470:Cell Survival; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics*; D004330:Drosophila / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D013696:Temperature; D014162:Transfection
D002130:Calcium Phosphates; D006360:Heat-Shock Proteins; D011993:Recombinant Fusion Proteins; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; C485838:calcium phosphate, monobasic, anhydrous; D004247:DNA; C020243:calcium phosphate; D015500:Chloramphenicol O-Acetyltransferase; C485829:calcium phosphate, dibasic, anhydrous