| 摘要 |
<p>PURPOSE:To enable to electrically fuse yeast cells with a high yield by dispersing yeast cells of different kinds in a fusion buffer solution containing electrolytes, collecting the cells by applying gravity acceleration, then applying a direct-current voltage. CONSTITUTION:A fusion buffer solution (A) is prepared by adding 0.5 to 0.9M of a nonelectrolyte such as glucose and 0.1 to 1mM of Ca salt or the like. Then, (C) a mixture of the solution A and Saccharomyces cerevisiae SH11509 strain and SH1512 strain (B) is introduced into the cylinder 15 of the centrifugal fusion chamber 2 provided with the mesh electrode 16, the bottom electrode 17, the connector 13, etc., in the cell fusion unit 1. Then the cylinder 15 is sealed with a cap 12, placed on the rotor 3 and fixed to the connector 13 and a voltage is applied by means of the operation panel 5. The rotor 3 is rotated to apply the gravity acceleration higher than 800G for more than three minutes to collect the cells between the electrodes 16 and 17. Then, direct current pulses are applied between the electrodes 16 and 17 to load the direct current voltage of 3.0 to 5.5kV/cm to the collected cells whereby the cells contacting with one another in a point are electrically fused with the cell membrane compression force.</p> |
