Effects of pH, Ca2+, temperature, and protease pretreatment on interkingdom fusion.
The incubation of carrot protoplasts and cultured Xenopus cells in a protease solution has been shown to enhance their subsequent interkingdom fusion by a high pH/high Ca2+ method. The effects of Ca2+ concentration, pH, and temperature on the frequency of heterokaryon formation have also been studied. Potentially viable heterokaryons have been repeatedly produced at high frequencies (consistently greater than 10%), far exceeding those so far achieved in PEG-mediated fusion. Cell aggregates are readily dispersed after this method of fusion, permitting the accurate estimation of fusion frequencies.
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002459:Cell Fusion / Q000187:drug effects*; D006863:Hydrogen-Ion Concentration; D010447:Peptide Hydrolases / Q000494:pharmacology*; D014675:Vegetables; D014981:Xenopus
D010447:Peptide Hydrolases; D002118:Calcium