Kjer K M, Fallon A M
Department of Entomology, University of Minnesota, St. Paul 55108.
Arch Insect Biochem Physiol. 1991;16(3):189-200. doi: 10.1002/arch.940160305.
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.
利用热休克蛋白-氯霉素乙酰转移酶（hsp-CAT）重组质粒的瞬时表达来确定影响使用DNA-磷酸钙共沉淀物转染培养的蚊细胞的参数。磷酸钙方法的效率受到受体细胞生长状态以及制备共沉淀物时温度的强烈影响。在最佳条件下，包括在50摄氏度形成DNA-磷酸钙共沉淀物，转染频率比使用先前描述的聚凝胺方法获得的频率高十倍。
