CAJ | 학술논문

目的观察外源基因在猪血管内皮细胞及小鼠体内表达情况.方法体外培养猪血管内皮细胞,脂质体转染法将pcDNA3-ICAM-2-Enhancer CD59cDNA转染猪的血管内皮细胞,以未转染的猪血管内皮细胞为阴性对照,人血细胞为阳性对照,经G418筛选(浓度为100 mg/L)获得具有抗性的克隆,通过流式细胞仪检测转染细胞的表达情况.通过显微注射将CD59基因片段导入小鼠授精卵,对出生的小鼠进行基因检测.结果 pcDNA3-ICAM-2-Enhancer-CD59cDNA重组质粒在细胞中表达强度为63.1%;转CD59基因小鼠外周血单核细胞(PBMCs)表达强度为73.38%.结论以人细胞间黏附因子Ⅱ(ICAM-2)为启动子并带增强子的CD59重组基因在猪的血管内皮细胞及小鼠体内均高效特异表达.
목적 관찰외원기인재저혈관내피세포급소서체내표체정황.방법 체외배양저혈관내피세포,지질체전염법장pcDNA3-ICAM-2-Enhancer CD59cDNA전염저적혈관내피세포,이미전염적저혈관내피세포위음성대조,인혈세포위양성대조,경G418사선(농도위100 mg/L)획득구유항성적극륭,통과류식세포의검측전염세포적표체정황.통과현미주사장CD59기인편단도입소서수정란,대출생적소서진행기인검측.결과 pcDNA3-ICAM-2-Enhancer-CD59cDNA중조질립재세포중표체강도위63.1%;전CD59기인소서외주혈단핵세포(PBMCs)표체강도위73.38%.결론 이인세포간점부인자Ⅱ(ICAM-2)위계동자병대증강자적CD59중조기인재저적혈관내피세포급소서체내균고효특이표체.
Objective To investigate the expression of human CD59 protein in transgenic pig aortic endothelial eeUs (PAECs) and mice. Methods The PAECs were cultured in vitro. The pcDNA3ICAM-2-Enhancer-CD59cDNA was transfected into the PAECs by lipofectinnmine mediated method. Normal PAECs served as negative control and human PBMCs as positive control. Stable transfectants were selected using G418 ( concentration 100 rag/L). The CD59 protein expression in transfected cells was as sayed by flow cytometry. CD59 gene fragments were transferred into mouse zygote by micro-injection and new born pups were detected for transgene and genotyped. Results The expression intensity of pcDNA3ICAM-2-Enhancer- CD59eDNA recombinant plasmid in PAECs was 63.1%. The expression intensity on the membrane of monocytes in periphery blood of transgenie mice of CD59 gene was 73.38%. Conclusion Recombinant CD 59 protein using ICAM-2 promoter with enhancer can be exlSt:essed highly, effectively and specifically in PAECs and transgenic mice.