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中国农学通报

中国农学通报 ›› 2014, Vol. 30 ›› Issue (2): 39-44.doi: 10.11924/j.issn.1000-6850.2013-1619

所属专题： 生物技术

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

家蚕BmN细胞S期同步化体系建立

黄蕾 江千令 阚云超 李丹丹   

  • 收稿日期:2013-06-13 修回日期:2013-07-11 出版日期:2014-01-15 发布日期:2014-01-15
  • 基金资助:
    国家自然科学基金“家蚕非编码RNA Bm-15在Notch信号通路及发育过程中的功能研究”(31100938)；国家自然科学基金“调控家蚕发育非编码RNA(non-coding RNA，ncRNA)的功能解析”(31172158)。

Establishment of S Phage Synchronization system for BmN Cells

  • Received:2013-06-13 Revised:2013-07-11 Online:2014-01-15 Published:2014-01-15

摘要： 细胞S期为DNA复制期，探索细胞S期的同步化将为研究各种调控因子对DNA复制、细胞分裂和增殖的影响奠定基础。本研究以家蚕BmN细胞为研究对象，采用脱氧胸苷(TdR)双阻断、脱氧胸苷-诺考达唑(TdR-Nocodazole)双阻断及羟基脲(HU)法，探索BmN细胞S期同步化的最佳体系。结果发现，TdR双阻断法处理对BmN细胞S期的同步化效率为67.2%，TdR-Nocodazole双阻断法处理为69.7%，HU处理对S期同步化效果最好，达90%以上。进一步对HU处理后不同释放时间点的细胞进行研究发现，1 mmol/mL HU处理后释放7 h时BmN细胞S期同步化效率最高，达94.9%。通过不同处理方法研究家蚕BmN细胞的S期同步化效果，发现1 mmol/mL的HU处理16 h，然后于正常培养基中释放7 h，可有效的将BmN细胞同步化在S期，且该处理方法简单，易于操作。

关键词: 品种, 品种

Abstract: S phase of cells is the DNA replication phase, exploring the synchronization of S phase will build foundation for the studies of various factors on the regulation of DNA replication, cell division and proliferation. In this work, three methods (TdR double occlusion, TdR-Nocodazole double occlusion and hydroxyurea occlusion) were used to study the S phage synchronization efficiency of BmN cells. The results showed that S phage sychronization efficiency of TdR double occlusion treatment was 67.2% , TdR-Nocodazole double occlusion was up to 69.7% , while hydroxyurea had the best sychronization efficiency with percentage more than 90%. Furthermore, the detailed conditions of HU occlusion were analyzed. The results showed that the highest S phase synchronization efficiency of BmN were up to 94.9% after 16 h treatment by 1 mmol/mL HU, following by being released in normal medium for 7 h. The results of different methods used to study the S phage synchronization efficiency of BmN cells showed that treatment of 1 mmol/mL HU for 16h and then were released in normal medium for 7h was the best synchronization method for BmN cells with its simple and easy operation.