解放军医药杂志Issue(5)：22-26,5.DOI:10.3969/j.issn.2095-140X.2014.05.006
携带LMO3基因逆转录病毒载体的构建及其在NIH/3 T3细胞中的表达
Construction of Recombinant Retroviral Vector Carrying LMO3 Gene and Its Expression in NIH/3T3 Cells
摘要
Abstract
Objective To construct recombinant retroviral vector carrying LIM domain only 3 (LMO3) gene and study its expression in NIH/3T3 cells. Methods The retroviral vector pLXSN-LMO3 was identified by restriction en-zyme analysis and DNA sequencing analysis, and then was transfected into retrovirus packaging cell line pA317, and G418 was used to select stable virus-producing cell lines. The recombinant retrovirus was used to infect NIH/3T3 cells in vitro, and the value of viral titer was determined. The NIH/3T3 cells were divided into three groups:experimental group (infected with the pLXSN-LMO3), negative control group (infected with the pLXSN) and control group. The expression of LMO3 in NIH/3T3 cells was identified by the immunofluorescence histochemistry and Western blot methods. Results The pLXSN-LMO3 recombinant retroviral vector had been constructed correctly by restriction enzyme analysis and se-quencing analysis, and the value of titer assayed on NIH3T3 cells was up to an average of 4. 04 × 106 cfu/ml. LMO3 al-bumen expression was detected in NIH/3T3 cells using immunofluorescence histochemistry and Western blot methods, and LMO3 was highly expressed in experimental group, and the differences in LMO3 expression were statistically signifi-cant when compared with those in negative control group and control group (P<0. 05). Conclusion The recombinant retroviral vector carrying LMO3 gene has been constructed successfully, which can highly express LMO3 in NIH/3T3 cells and has potential utility in further gene therapy.关键词
单纯LIM蛋白3/NIH/3T3细胞/逆转录病毒科感染/pLXSNKey words
LIM domain only 3/NIH/3T3 cell/Retroviridae infection/pLXSN分类
医药卫生引用本文复制引用
罗芸,张新宇,万东君,刘晓花,付学锋..携带LMO3基因逆转录病毒载体的构建及其在NIH/3 T3细胞中的表达[J].解放军医药杂志,2014,(5):22-26,5.基金项目
甘肃省自然科学基金资助项目(1107RJZA106) （1107RJZA106）
