CAJ | 학술논문

目的：构建 EGFP转基因小鼠及建系,并观察转基因小鼠的细胞及组织荧光发出状况。方法使用 Gateway clone技术构建 eEGFP载体,通过DNA显微注射法将构建好的 EGFP 载体注入受精卵内,转移至假孕母鼠内生出转基因小鼠。将转基因小鼠后代中的阳性小鼠互交,逐步淘汰杂合子小鼠。饲养过程测量小鼠的基本数据。培养转基因小鼠的脊髓源神经干细胞,并取部分常用组织石蜡切边,在荧光显微镜下观察。结果成功构建了 EGFP 转基因小鼠,并在屏障环境下饲养繁殖至7代。与野生型小鼠相比,EGFP转基因小鼠基本数据无明显差异。各常用组织切片在荧光显微镜下均可观察到绿色荧光,从脊髓中培养获得的神经干细胞也可观察到绿色荧光。结论 EGFP小鼠组织在常规处理后可观察到发出绿色荧光的细胞；小鼠脊髓源神经干细胞在体外培养3代仍可观察到绿色荧光,可利用这种自然标记的细胞研究神经干细胞在脊髓损伤修复中所产生的作用。
목적：구건 EGFP전기인소서급건계,병관찰전기인소서적세포급조직형광발출상황。방법사용 Gateway clone기술구건 eEGFP재체,통과DNA현미주사법장구건호적 EGFP 재체주입수정란내,전이지가잉모서내생출전기인소서。장전기인소서후대중적양성소서호교,축보도태잡합자소서。사양과정측량소서적기본수거。배양전기인소서적척수원신경간세포,병취부분상용조직석사절변,재형광현미경하관찰。결과성공구건료 EGFP 전기인소서,병재병장배경하사양번식지7대。여야생형소서상비,EGFP전기인소서기본수거무명현차이。각상용조직절편재형광현미경하균가관찰도록색형광,종척수중배양획득적신경간세포야가관찰도록색형광。결론 EGFP소서조직재상규처리후가관찰도발출록색형광적세포；소서척수원신경간세포재체외배양3대잉가관찰도록색형광,가이용저충자연표기적세포연구신경간세포재척수손상수복중소산생적작용。
Obj ective To establish transgenic mice expressing eEGFP gene and transgenic mouse lin-eages,and to observe fluorescence from tissues and cells of transgenic mice were observed by fluorescence meanwhile.Methods Gateway clone technology was used to construct the eEGFP carrier which was infec-ted by DNA Microinj ection into fertilized ovum,which was then transferred into pseudopregnancy mice to give birth to transgenic mice.We intercrossed the descendants and gradually eliminated heterozygote mice. We measured the basic data in the feeding process.We cultivated cells of transgenic mice,made paraffin sections and observed them under fluorescence microscope.Results We constructed EGFP transgenic mice to the seventh generation.Compared with wild type mice,there were no obvious differences in the basic data of EGFP transgenic mice.Green fluorescence of culture cell and tissue sections was observed under fluorescence microscope.Conclusion It can be observed Green fluorescence of EGFP mouse cell culture in vitro till the third generation,which may be applied to study the role of neural stem cells in the repair of spinal inj ury.