摘要
目的构建稳定表达Flt3分子的转基因细胞株。方法将Flt3基因重组入逆转录病毒载体pEGZ-term,通过293T细胞的包装获得具有感染能力的完整重组病毒载体,进而采用“一箭双星法”即同时感染BaF和L929细胞,构建Flt3转基因细胞株。结果成功获得稳定表达Flt3的转基因细胞株BaF/Flt3和L/Flt3。结论采用“一箭双星法”成功构建Flt3转基因细胞株,为以Flt3为靶分子的研究奠定了物质基础,也为类似的高难度的转基因细胞株的构建提供了有价值的经验。
Objective To construct transgenic cell line that stably expressing Fh3. Methods Fh3 gene was subcloned into retroviral expressing vector pEGZ-term. Whole combinated retroviral expressing vector was integrated by package cell; 293T cell, and then Fh3 transgenic cell line was constructed by transfecting BaF Cells and L929 cells at same time. So this new strategy was named “kill two birds with one stone”. Results Transgenic cell lines those stably expressing Fit3, including BaF/Fh3 and L/Flt3, were constructed successfully. Condusion Fh3 transgenic cell lines were established successfully. It provided valuable tools to study Fh3 biological functions. The“ kill two birds with one ston” strategy provided an effective solution to the low efficiency of transgenic cell construction of the suspending cell.
出处
《中国血液流变学杂志》
CAS
2005年第3期344-347,共4页
Chinese Journal of Hemorheology
基金
国家自然基金青年科学基金项目(30400394)
江苏省放射医学与防护重点实验室开放经费资助项目(KJS05031)
