水动力转染鸡PPARγ基因在小鼠肝脏中的表达  

Expression of chicken PPARγ gene by hydrodynamics-based transfection in mouse liver

作　　者：王启贵[1] 王娉[1] 庄淑珍[1] 郑耀虎[1] 李辉[2] 张富春[1] 

机构地区：[1]新疆大学新疆生物资源基因工程重点实验室,新疆乌鲁木齐830046 [2]东北农业大学动物科学与技术学院,黑龙江哈尔滨150030

出　　处：《扬州大学学报（农业与生命科学版）》2007年第1期26-29,共4页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：中国博士后科学基金资助项目(2005037018)

摘　　要：采用水动力转染技术将含有家鸡PPARγ基因的真核表达质粒、表达该基因siRNA的干扰质粒注射到小鼠体内,并利用RT-PCR方法检测PPARγ基因在小鼠脏器中的表达情况,以建立检测重组质粒瞬时表达和筛选siRNA序列的有效方法。结果表明:注射重组质粒pcDNA3/PPARγ后家鸡PPARγ基因在小鼠的肝脏中高效表达;同时注射重组质粒pcDNA3/PPARγ和表达该基因siRNA序列的干扰质粒pGenesil-1/PPARγshRNA1后,家鸡PPARγ基因在小鼠肝脏中的表达明显地受到抑制。与传统的真核细胞转染技术相比,水动力转染技术具有快速、简单、方便、高效、安全、成本低等优点,可作为外源重组DNA表达的检测和siRNA的筛选等一种有效的方法。In order to establish an efficient method for detecting recombination DNA expression and screening siRNA sequence, hydrodynamics-based transfection was studied in mouse. The eukaryotic expression plasmid pcDNA3/PPARγ containing chicken PPARγ gene and interference plasmid pGenesil-1/PPARγshRNA1 with siRNA of PPARγ gene were transfected into mice by Hydrodynamics based transfection. Then the expression of PPARγ gene was detected by RT-PCR in mouse viscus. The chicken PPARγ gene was highly expressed in mouse liver tissue after injecting pcDNA3/PPARγ, and expression of the chicken PPARγ gene was significantly suppressed in mouse liver tissue after injecting the plasmid mixture of pcDNA3/PPARγ and pGenesil-1/PPARγshRNA1. Hydrodynamics-based transfection is a novel developed gene transfer technique in vivo, and has lots of advantages, such as convenience, simplicity, high efficiency, safety, and low cost, etc. compared with the traditional eukaryocyte transfection techniques, and can be used as a valuable tool for detecting recombination DNA expression and screening siRNA in live animals.

关 键 词：小鼠 PPARΓ基因 水动力转染 表达 

分 类 号：Q786[生物学—分子生物学]