FGFR2 mRNA 3'UTR萤光素酶报告载体的构建及活性检测

o obtain Constructed the overall length and difference truncations of FGFR2 3'UTR- luciferase reporter vector. For the study of the regulation of FGFR2 in transcription and translation lever in esophageal tumor microenvironment provided a useful tool Methods through extracting mRNA from esophageal cancer associated fibroblasts, reverse transcribed into cDNA, template for PCR amplification of overall length and truncated FGFR2 3 'UTR sequence and then made these sequences cloned into psi-CHECK-2 vector. To constructed the overall length and difference truncations of FGFR2 3'UTR- luciferase reporter vector. After sequencing facility showed correct then the luciferase reporter vector and miRNA mimics were transferred into 293T cells. after 48h test the luciferase activity, determine the miRNA and targeting the function of target genes.Results Successful build the overall length psi - CHECK -2 FGFR23 'UTR and different truncated psi - CHECK - 2-3' UTR FGFR2 luciferase report system and test results show ed that miR - 671-5 p combined with FGFR2 3 'UTR . Conclusion Successful construction FGFR2 mRNA 3 'UTR luciferase report vector. And preliminary evidence that miR - 671-5 p combined with FGFR2 have targeting effect.