Transfection of cultured cells of the cotton boll weevil, Anthonomus grandis, with a heat-shock-promoter-chloramphenicol-acetyltransferase construct.
Expression of heat shock proteins (hsp) in the BRL-AG-3C cell line from the cotton boll weevil was examined. It was determined that the maximal expression of endogenous hsp occurred at 41 degrees C. Various transfection methods were then compared using this cell line in conjunction with a transiently expressed bacterial gene marker (chloramphenicol acetyltransferase) which was under the control of the Drosophila hsp 70 gene promoter. The cationic lipid preparation Lipofectin was found to be very efficient at transfecting the boll weevil cells. Polylysine and 20-hydroxyecdysone-conjugated polylysine were moderately effective, whereas polybrene and electroporation, under the conditions reported herein, were ineffective at transfecting this cell line.
Stiles
B
B
USDA, ARS, Bioscience Research Laboratory, Fargo, North Dakota 58105.
Heilmann
J
J
Sparks
R B
RB
Santoso
A
A
Leopold
R A
RA
D000818:Animals; D002478:Cells, Cultured; D001517:Coleoptera / Q000235:genetics*; D004441:Ecdysterone / Q000235:genetics; D005822:Genetic Vectors; D006360:Heat-Shock Proteins / Q000235:genetics*; D006583:Hexadimethrine Bromide / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000235:genetics; D011107:Polylysine / Q000235:genetics; D011401:Promoter Regions, Genetic; D014162:Transfection
D006360:Heat-Shock Proteins; D010714:Phosphatidylethanolamines; D011107:Polylysine; D006583:Hexadimethrine Bromide; D004441:Ecdysterone; C020888:1,2-dielaidoylphosphatidylethanolamine