CAJ | 학술논문

目的为明确EC-SOD在动脉粥样硬化发生发展中的作用,课题组体外构建pCDNA 3.1/EC-SOD真核表达载体,并观察其在THP-1单核源性巨噬细胞中的表达情况。方法重组质粒经PCR、酶切以及序列分析正确无误后,借助Lipofectamine 2000将转染到THP-1单核源性巨噬细胞中,24h后倒置荧光显微镜下观察转染情况。结果重组质粒经酶切鉴定、PCR和测序分析后,证实pCDNA 3.1/EC-SOD质粒构建成功。将pCDNA 3.1/EC-SOD通过脂质体转入THP-1单核源性巨噬细胞后,倒置荧光显微镜下观察转染效率。结论 pCDNA 3.1/EC-SOD载体构建成功,并成功在THP-1单核源性巨噬细胞中表达,为研究EC-SOD在动脉粥样硬化发生发展中的作用奠定了基础。
목적위명학EC-SOD재동맥죽양경화발생발전중적작용,과제조체외구건pCDNA 3.1/EC-SOD진핵표체재체,병관찰기재THP-1단핵원성거서세포중적표체정황。방법중조질립경PCR、매절이급서렬분석정학무오후,차조Lipofectamine 2000장전염도THP-1단핵원성거서세포중,24h후도치형광현미경하관찰전염정황。결과중조질립경매절감정、PCR화측서분석후,증실pCDNA 3.1/EC-SOD질립구건성공。장pCDNA 3.1/EC-SOD통과지질체전입THP-1단핵원성거서세포후,도치형광현미경하관찰전염효솔。결론 pCDNA 3.1/EC-SOD재체구건성공,병성공재THP-1단핵원성거서세포중표체,위연구EC-SOD재동맥죽양경화발생발전중적작용전정료기출。
To investigate the ef ect of EC-SOD in development of atherosclerosis (As), we recombination vector of pCDNA 3.1/EC-SOD in vitro and to observe its expression in monocyte-derived macrophages. METHODS EC-SOD fragment were cloned and linked to plasmid pCDNA 3.1 in vitro. After plasmid polymerase chain reaction (PCR), Restriction enzyme digestion and sequencing to confirm the vector of EC-SOD was successful y constructed, with the help of Lipofectamine 2000 we transfected the vector to macrophages, and we detected the transfection ef iciency with fluorescence. CONCLUSION The vector of EC-SOD was successful y constructed and its working in macrophages, this work was a great base for study the role of EC-SOD in development of As.