Mg2+和Na+对多核酶系统体外切割反应的影响

田生礼,郑硕;刘士德;徐东平;TakaoOHNUMA

中国生物化学与分子生物学报 ›› 2008, Vol. 24 ›› Issue (07) : 612-618.

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中国生物化学与分子生物学报 ›› 2008, Vol. 24 ›› Issue (07) : 612-618.
研究论文

Mg2+和Na+对多核酶系统体外切割反应的影响

  • 田生礼1) *, 郑 硕1), 刘士德1), 徐东平2), Takao OHNUMA3)
作者信息 +

Effects of Divalent Mg2+ and Monovalent Na+ on Cleavage
Reactions by Multiribozyme System in vitro

  • <SPAN lang=EN-US style=
Author information +
文章历史 +

摘要

为了进一步了解2价Mg2+和1价Na+存在与否的情况下,多核酶系统对底物RNA的切割效率,构建了pGEMCoat′A,pGEMCoat′A196Rz 质粒和pGEMMDR1靶质粒.通过用SP6/T7转录试剂盒在体外转录RNA, 在无细胞系统进行切割反应,反应产物通过6%变性聚丙烯酰胺凝胶电泳,干胶、X光片曝光自显影,利用Image J 生物图像分析软件分析. 结果表明,多核酶系统的切割效率依赖于二价Mg2+的浓度,切割产物随Mg2+浓度的增加而增加,而且具有反应时间的依赖性. 在Na+浓度低于200 mmol/L且单独存在时,没有切割产物生成.相反,在Na+和Mg2+共存时,表现出Na+抑制Mg2+诱导的切割活性,切割效率明显低于Mg2+单独存在时的结果.这些结果提示,在生理环境下,Mg2+对于多核酶系统对底物的切割反应是必需的,而Na+则不是.

关键词

多核酶系统 / Mg2+ / Na+ / RNA切割 / 切割反应

Key words

multiribozyme system / divalent Mg2+ / monovalent Na+ / RNA cleavage / cleavage reaction

引用本文

导出引用
田生礼,郑硕;刘士德;徐东平;TakaoOHNUMA. Mg2+和Na+对多核酶系统体外切割反应的影响[J]. 中国生物化学与分子生物学报, 2008, 24(07): 612-618
<SPANlang=EN-USstyle=. Effects of Divalent Mg2+ and Monovalent Na+ on Cleavage
Reactions by Multiribozyme System in vitro
[J]. Chinese Journal of Biochemistry and Molecular Biology, 2008, 24(07): 612-618

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