CXCR4启动子报告载体的构建

摘要

目的：构建并鉴定含人 CXCR4基因启动子的荧光素酶的报告载体。方法采用克隆人 CXCR4基因启动子的序列，构建报告载体 PgL4.17-CXCR4，以其转染 Jurkat 细胞，然后测定荧光素酶的活性。结果扩增得到的 CXCR4启动子序列与克隆人 PgL4.17-CXCR4的 CXCR4启动子序列和 GenBank 报道的一致。实验组荧光素酶的表达活性是874809.0200±39510.6246，与空白组的465.9000±26.2501和对照组的37981.9800±2384.3412总体比较差异有统计学意义（F =4678.919，P <0.001）。结论按照本实验方案，含人 CXCR4基因启动子的荧光素酶的 PgL4.17-CXCR4报告载体被成功构建。%Objective To construct and identify a luciferase reporter vector PgL4. 17-CXCR4 containing human CXCR4 promoter. Methods The sequence of CXCR4 promoter gene was cloned,and was inserted into the reporter vector PgL4. 17 to construct luciferase reporter vector PgL4. 17-CXCR4 ,the recombinant plasmids were transfected into Jurkat cells. The luciferase activity was evaluated. Results CXCR4 promoter gene fragment was amplified by PCR. The insertion sequence of CXCR4 promoter in the experimental group was 874 809. 020 0 ± 39 510. 624 6,was 465. 900 0 ± 26. 250 1 in the blank group,and was 37 981. 980 0 ± 2 384. 341 2 in the control group( F =4 678. 919,P < 0. 001). Conclusion Luciferase reporter vector PgL4. 17-CXCR4 containing human CXCR4 pro-moter can be constructed successfully.