摘要
目的 建立 HRX- EEN融合基因转基因小鼠为新药的筛选提供理想的动物模型。 方法 通过拼接 ,用 3个不同来源的 DNA构建成 HRX- EEN融合基因 ,并将其装入 h CG转基因载体中 ;通过显微注射将上述 DNA导入受精卵雄核中 ,植入养育母鼠输卵管 ,发育获得 G0 代小鼠 ;PCR检测融合基因的整合 ,逆转录 -聚合酶链反应 (reverse transcription- polymerase chain reaction,RT- PCR)检测融合基因的表达。结果 测序证实拼接的 HRX- EEN融合基因正确 ;PCR共检测出 7个 HRX- EEN转基因阳性的 G0 代小鼠 ,将这些小鼠与野生型的 C5 7系小鼠交配 ,结果除 35号死亡外 ,其余 G0 代转基因阳性小鼠均产下 F1代 ,建成 6个单系的 HRX- EEN转基因小鼠。用 RT- PCR方法检测其中 5个系小鼠 HRX- EEN融合基因的表达状况 ,证实该融合基因在 3个单系中稳定表达。表达融合基因的转基因小鼠迄今未发现明显异常。 结论 构建完成的 HRX- EEN转基因小鼠模型能稳定表达。
Objective To study the biological function of fusion gene HRX-EEN and its role in leukemogenesis, and to provide an ideal animal model for anti-leukemia drug screening. Methods HRX-EEN fusion gene was constructed by use of three different DNA fragments, and it was inserted into hCG transgenic vector. G 0 transgenic mice were obtained by microinjection of the recombined DNA into the pronucleus of zygotes, followed by implantation of the injected zygotes into pseudopregnant mice. The integration of the transgene was tested by PCR and its expression by reverse transcription-polymerase chain reaction(RT-PCR). Results The sequence of recombined HRX-EEN gene was confirmed by sequencing. PCR testing revealed a total of 7 G 0 transgenic mice, these mice were then mated with C57 wild type mice. Except mouse No. 35 that died, the others all had their F1 offsprings. From these 6 lines of transgenic mice, HRX-EEN gene was found to be stably expressed in 3 lines by RT-PCR. Up to now, all transgenic mice expressing the fusion gene have no obvious abnormal phenotypes. Conclusion A transgenic mice model in which the HRX-EEN fusion gene can be stably expressed has been established.
出处
《中华医学遗传学杂志》
CAS
CSCD
2003年第6期522-527,共6页
Chinese Journal of Medical Genetics
基金
国家自然科学基金(39970403)
国家杰出青年基金(39925023)
上海市科技发展基金(01DJ14002
99XD14005)
上海市高等学校科学技术发展基金项目(99B11)~~
