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Efficient transfection of mosquito cells is influenced by the temperature at which DNA‐calcium phosphate coprecipitates are prepared
Author(s) -
Kjer Karl M.,
Fallon Ann Marie
Publication year - 1991
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.940160305
Subject(s) - transfection , calcium , phosphate , biology , recombinant dna , dna , microbiology and biotechnology , plasmid , biochemistry , chloramphenicol , hsp70 , heat shock protein , biophysics , chemistry , gene , organic chemistry , antibiotics
Trnsient expression of a heat‐shock protein‐chloramphenicol acteyltrans‐Perase (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°C, transfection frequencies were up to tenfold higher than those obtained using the previously described polybrene procedure.