高圧窒素ボンベによる肝細胞核及び腹水肝癌細胞核の破砕とRNAポリメラーゼ活性におよぼす影響

細胞核の破砕方として高圧窒素ガスボンベを用いて,その破砕効果を検討した.ラット肝細胞核を窒素で加圧,解除によって破砕した. 3mM MgCl(2)の存在下では末破壊核,核小体が残った. 0.02mM MgCl(2)存在下ではこれらは完全に破壊された. AH-130細胞核をRNAポリメラーゼ抽出のために超音波処理すると,RNAポリメラーゼは失活した. 他方,同様の条件下で高圧窒素ガスの加圧及び解除によって失活することなくRNAポリメラーゼを抽出した.

Nitrogen pressure bomb was used for the disruption of nuclei. The effects of disruption were checked by electron microscopy and estimtion of released RNA-polymerase. Disruption of rat liver nuclei was incomplete in the presence of 3mM Mg(++) and complete in the presence of 0.02mM Mg(++), in which nuclear membrane and nucleoli were completely disrupted. For releasing RNA polymerase from AH-130 nuclei, disruption of nuclei was done by sonication in the presenec of 0.3M ammonium sulphate. RNA-polymerase activity was lost makedly by sonication. In the same condition AH-130 nuclei was disrupted by nitrogen pressure bomb, in which RNA polymerase activity was not lost and it was recovered in the supernatant fractino with the yield of 90%.