摘要
目的:观察能否应用高压注射体内转染法实现外源DNA在肝脏中高效表达,为嗜肝病毒分子生物学研究奠定方法学基础。方法:将大容量乙肝病毒表面抗原表达质粒pVAX1/SDNA溶液,通过尾静脉快速注射入小鼠体内,用免疫组化检测小鼠心、肺、肝、脾、肾组织中HBsAg的表达情况。结果:经小鼠尾静脉高压注射pVAX1/S质粒DNA后,HBsAg主要在肝脏表达;且高压注射组明显强于缓慢注射组;高压注射5μgpVAX1/S质粒DNA后8小时,在肝脏观察到较强的HBsAg表达。结论:通过尾静脉高压注射,能实现HBsAg主要在肝脏高效表达。
Objective： To investigate whether an efficient expression of exogenous DNA in liver can be achieved by intravenous injection of DNA using a hydrodynamics- based in vivo transfection method, so as to provide the methodological bases for molecular biological studies of hepadnavirus. Method： Mice were rapidly injected with a large volume of pVAX1/S plasmid DNA solution via tail vein. The expression of HBsAg in internal organs including heart, lung, liver, spleen and kidney was detected by immunohistochemical technique. Result： The highest HBsAg expression level was observed in liver by systemic administration of pVAX1/S DNA using a hydrodynamics - based procedure. The HBsAg expression by hydrodynamics - based procedure was significantly higher than that by slow injection method. Eight hours after injection of 5μg pVAX1/S by hydrodynamics - based method, an obvious HBsAg expression was detected. Conclusion： Using hydrodynamics- based method, an efficient expression of HBsAg in hepatocytes can be achieved.
出处
《华西医学》
CAS
2005年第4期697-699,共3页
West China Medical Journal
基金
国家杰出青年科学基金(编号30325036)。
关键词
转染
高压注射体内转染
免疫组化
transfection
hydrodynamics- based transfection
immunohistochemistry
