摘要
目的HVEM突变体和EGFP共表达复制缺陷型腺病毒载体的构建与鉴定。方法RT-PCR获得人HVEM的cDNA,使其与LIGHT分子结合的表位点突变,保留与BTLA结合的关键位点。构建经修饰的HVEM基因的复制缺陷型腺病毒载体Adv-mHVEM-IRES2-EGFP并对其进行鉴定和转染效率测定。结果成功构建HVEM突变体和EGFP复制缺陷型腺病毒载体,经过测序与预期一致。该载体能够表达突变的HVEM和EGFP蛋白,体外培养细胞转染效率达到95%以上。结论构建的腺病毒载体能够共表达目的基因的产物HVEM突变体和EGFP蛋白,对体外细胞具有高效转染能力。
Objective Construction and identification of Adv-mHVEM-IRES2-EGFP.Methods HVEM cDNA bind domain of LIGHT was mutated by RT - PCR and PCR. Adenovirus expression vector Adv-mHVEM-IRES2-EGFP was constructed with mHVEM and EGFP by means of AD-Easy system.The vector was identified and evaluated the efficiency of transfecfion. Results HVEM and EGFP were expressed in the culture cells transfected by the vector whir a transfection efficiency over 95 %. Conclusion The recombinate vector Adv-mHVEM-IRES2-EGFP could express the protein of mHVEM and EGFP in cell line with high transfection efficiency.
出处
《四川医学》
CAS
2006年第7期668-670,共3页
Sichuan Medical Journal
基金
国家自然科学基金资助项目(30571863)
