氟化物对限制性内切酶酶切活性改变的研究

Study on the Effect of Fluoride on Restriction Endonuclease Activity

目的 :探讨氟化物对限制性内切酶酶切作用改变的影响。方法 :常规方法提取纯化PBR32 2质粒 ;将NaF与PBR32 2质粒 37℃共同作用 1h ,再用限制性内切酶BamH1 对PBR32 2质粒进行酶切 3h ,同时将NaF作用的PBR32 2用层析柱纯化后再进行酶切反应 ;将酶切产物进行电泳 ,观察结果并拍照。结果 :随着NaF浓度的增加 ,BamH1 对PBR32 2的酶切作用减弱 ,当加入 5mmol LNaF时 ,BamH1 几乎无酶切活性 ,而对加入 5mmol LNaF的PBR32 2用层析柱进行纯化 ,以除去NaF ,再对其进行酶切 ,可完全切开。而且氟离子可导致酶切片段在电泳中显示拖尾现象。结论

Objective To explore the effect of fluoride on the restriction endonuclease.Methods The PBR322 plasmid was amplified by using normal molecular techniques and incubated with NaF for 1 h at 37℃.After that,it was enzymed by BamH 1.The PBR322 plasmid incubated with NaF was purified and then enzymed by BamH 1.Results The enzyming effect of BamH 1 on PBR322 was decreasing when the NaF concentration increased.However,when the PBR322 was purified,its enzyming effect by BamH1 did not change.The fluoride induced the DNA cleavage phenomenon of enzymed fragments.Conclusion The fluoride could inhibit the activity of restriction endonuclease. China J Cancer Prev Treat,2003,10(5):463-464