家蚕BmN细胞S期同步化体系建立  

Establishment of S Phage Synchronization System for BmN Cells

作　　者：黄蕾[1] 江千令[1] 阚云超[1] 李丹丹[1] 

机构地区：[1]中英.南阳洛桑昆虫生物学联合实验室/南阳师范学院,河南南阳473061

出　　处：《中国农学通报》2014年第2期39-44,共6页Chinese Agricultural Science Bulletin

基　　金：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)

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。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.

关 键 词：细胞同步化 S期 羟基脲 

分 类 号：Q962[生物学—昆虫学]