中文摘要：

目的构建miR-638慢病毒过表达载体,探讨其对MAPK14基因的靶向调控作用。方法分别构建pSicoR-miR-638及pMIR-Report-MAPK14过表达载体,应用双荧光素酶报告基因活性、Western blot及实时荧光定量PCR（qRT-PCR）法验证miR-638与MAPK14的靶向调控关系。结果经PCR、酶切及测序结果证明成功构建了pSicoR-miR-638及pMIR-ReportMAPK14重组质粒,并通过实验证实miR-638过表达明能显抑制MAPK14的蛋白及mRNA表达（P〈0.05）;抑制miR-638的表达,则MAPK14蛋白及mRNA的表达水平明显上调（P〈0.05）。结论成功构建pSicoR-miR-638慢病毒载体,并证实其可通过靶向作用于MAPK14-3’UTR的特异序列而直接抑制MAPK14基因的表达。

英文摘要：

To construct a lentiviral vector of miR-638 and verify its targeted effect on MAPK14 gene, pSicoRmiR-638 and pMIR-Report-MAPK14 over-expression vectors were constructed. Then the targeted effect of miR-638 on MAPK14 gene was verified by the relative luciferase activity, Western blot and real-time PCR（qRT-PCR）test. The PCR analysis, restriction enzyme digestion analysis and DNA sequencing analysis demonstrated that the recombinant plasmids pSicoR-miR-638 and pMIR-Report-MAPK14 3′-UTR were constructed successfully. Data also showed that over-expression of miR-638 cold suppress the mRNA and protein expression levels of MAPK14significantly（P 0.05）. In conclusion, the lentiviral vector containing miR-638 gene is constructed successfully,which can suppress MAPK14 gene expression by targeting the specific sequence of MAPK14-3′UTR.