核定位信号突变型P21真核表达载体的构建与其对HepG2_2_15细胞病毒复制的.pdfVIP

2010 26 4 387 P21 H epG2215 1, 2 1 1 1 邱荣元 , 何生松 , 陈 锋 , 庞 然 ( 1华中科技大学同济医学院附属协和医院肝病科, 武汉 430023; 2岳阳市 二人民医院消化内科, 湖南 岳阳 414000) P2 1 HBV P2 1, DNA , P2 1 pDsRed1- C1, pDsRed1- C1- p21NLS- , DNA , H epG2215, RT - PCR pDsRed1- C1- p21NLS- , , ELISA HB sAgH BeAg pDsRed1- C1- p21NLS- , , , (P 001) P21, H epG2215 P2 1, P2 1 ; , ; ; H epG2215 + R5126 2 A 1001- 5256( 2010) 04- 0387- 05 Construction of eukaryotic expression vector of P2 1mutation type by nuclear location signaland its effect on virus replication inH epG2215 Cells QIUR ong - yuan, HE Sheng - song, CHEN Feng, etal. (D ep artm ent of H ep atology, UnionH osp ital, ongj iM ed ical College, Huazhong University of Science and echnology, W uhan, 430022, China) Abstract: Objective T o construct the eukar otic expression vector of pDsRed 1- C1- p2 1NLS- gene and stud its func tion. M ethods The mutation w ith nuclear localization signal of p21w as obtained b site- directedm utagenesis. p2 1NLS - w as d irectl cloned into eukar otic expressive plasm id pDsRed1- C1 and transfected intoH epG2 Cells b liposom e- m ed iated DNA transfection. The p21 NLS- gene express ion, intracellu lar localization of the p21 and HBV antigens w ere detected b RT - PCR, fluorescence m icroscope and EL ISA, respectivel . esults The express ion of pDsRed1- C1- p2 1NLS- was m ain l localized in the c toplasm compared to pDsRed1- C1- p2 1WT wh ich w as localized in the nucleus. The expression of pDsRed1- C1- p21NLS- mi proved the rep lication ofH BV in H epG2. 2. 15. Conclusion The differ ent intracellular localization of p21 has various mi pacts on replication ofHBV in H epG2215 cells. K ey words: nuclear