人死亡受体5全长基因的构建及转染胶质瘤细胞-core.pdfVIP

25 5 2009 9 [ ] 1000-8861( 2009) 05-059 1-05 5 1, 2# 2 2 2 2 2 2 1* 庄国洪 , 黎之静 , 孟庆宇, 陈彩霞, 王 娟 , 李文珠 , 张佳锴 , 朱 迅 [ ] 5( death receptor 5, DR5) pcDNA 3. 1/DR5, pcDNA3. 1/ GFP/DR5, U 138 PCR DR5 , pcDNA 3. 1/GFP / DR5, pcDNA3. 1/DR5 , , 2 pcDNA 3. 1/GFP/DR5pcDNA 3. 1/GFP U 138, 72 h, ( RT-PCR ) DR5mRNA , DR5 An ti-DR5 U 138 DR5 , U 138, RT-PCR , U 138 DR5 mRNA, Anti-DR5 U 138 DR5 , DR5 , DR5 U 138 [ ] ; ; DR5; [ ] Q782 [] A Construction of full-length DR5 gene and its transfection into glioma cell line ZHUANG Guohong, LI Zh ijing, MENG Q ingyu, CHEN Cai ia, WANG Juan, LIW enzhu, ZHANG Jiakai, ZHU Xun D epartment of Immunology, School of BasicM edicalSciences, Jilin University, Changchun 130021, China [Abstract] Objective T o construct tw o eukaryotic e press ion vectors of hum an DR5 full length gene wh ich nam ed pcDNA 3. 1/ DR5 and pcDNA 3. 1/GFP/DR5, and observe the transfection efficiency after transfecting them into gliom a cell line U 138. M ethods Ful-l length DR5 gene w as constructed by us ing overlapping PCR techniques, for constructing recom binant p lasm ids of pcDNA 3. 1/DR5 and pcDNA3. 1/GFP/DR5. U tilizing liposom e transfection, pcDNA3. 1/DR5 and pcDNA 3. 1/GFP /DR5 recomb inant w ere transduced into U 138, respectively. The levels of DR5 mRNA w ere determ ined 72 h post transfection by relative quantitative RT- PCR, wh ile the e pressions of DR5 in U 138 cell w ere detected by flow cytom etry m ethod. Results The fu ll-length DR5 gene w as constructed successfu lly. The eukaryotic e pression vectors pcDNA 3. 1/DR5 and pcDNA 3. 1/GFP /DR5 w ere transfected into U 138, respectively. The e pression levels of DR5 mRNA and protein w ere s ignificantly increased after transfection. An ti-DR5 cou ld induce U 138 apop tosis obviously. Conclusion The optmi iz