可诱导心肌特异性表达TRX蛋白转基因小鼠模型的建立  被引量:1

Establishment of transgenic mice model with myocardium-specific expression of TRX protein

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作  者:孙煜 李成刚 周立[3] 

机构地区:[1]湖北省食品药品监督管理局药品审评认证中心,湖北武汉430071 [2]武汉市刚子科贸有限公司,湖北武汉430071 [3]武汉大学动物实验中心/ABSL-3实验室,湖北武汉430071

出  处:《海南医学》2014年第8期1093-1096,共4页Hainan Medical Journal

摘  要:目的建立心肌特异性、高效表达鼠源硫氧还蛋白-1(Thioredoxin 1,Trx-1)的转基因小鼠模型。方法将TRE-Tight-Trx-1与能够控制TRE-Tight下游基因在心肌特异性表达的α-MHC-rtTA-hGH转基因载体分别显微注射入C57BL/6小鼠受精卵细胞中得到的分别含有一段转基因载体的转基因小鼠,再将这两种转基因小鼠进行交配,获得同时含有TRE-Tight-Trx-1和α-MHC-rtTA-hGH这两段基因的双阳性子代转基因小鼠。使用强力霉素(Dox)持续诱导6周龄双阳性小鼠6周,再用Western blot方法检测转基因小鼠心肌细胞中Trx-1表达量。结果与野生型C57BL/6小鼠比较,经过强力霉素诱导的双阳转基因小鼠心肌细胞中Trx-1有高表达量(P<0.05)。结论我们得到了可诱导、高效表达Trx-1的转基因小鼠系,为心肌肥大疾病的研究和治疗提供新的研究思路。Objective To establish Thioredoxin 1 (Trx-1) transgenic mice model with myocardium-specifie high expression ofThioredoxin (Trx-1). Methods TRE-Tight-Trx-I and ct-MHC-rtTA-hGH gene vector which spe- cifically controls the expression of TRE-Tight downstream gene in cardiomyocytes were microinjected into the zygote of C57BL/6 mice respectively for establishing the transgenic mice with transgenic vectors. After mating, a double-pos- itive offspring with bothTRE-Tight-Trx-1 and α-MHC-rtTA-hGH gene were obtained. And then doxycycline was add- ed to feed the double positive transgenic mice aged over 6 weeks for 6 weeks, where the concentration of Trx-1 in car- diomyocytes was detected by Western Blot. Results The transgenic mice feeding with doxycycline showed high ex- pression levels of Trx-1 in cardiomyocytes compared with wild type C57BL/6 mice. Conclusion We established a Trx-1 transgenie mice model with inducible and high expression of Trx-1, which provided a new pattern of thinking in the research and therapy for cardiac hypertrophy.

关 键 词:硫氧还蛋白-l α-肌球蛋白重链 强力霉素 Tet-on/pTRE-Tight诱导表达系统 

分 类 号:R-332[医药卫生]

 

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