Objective: To build with wild type and mutant integrins beta 1(integrin beta 1

ITGβ1)-3'end of translation region (UTR) dual luciferase reporter gene expression system (DLR) (pMIR-ITGβ1-3' UTR)

through the dual luciferase report gene detects the kits ITGβ1

this study miR-29 ITGβ1 gene targeting regulatory role.Which based on regulating ITGβ1 transcription level after the screening of tanshinone. Method: Extraction of whole blood RNA

reverse transcriptase mRNA into cDNA

for the template

RT-PCR amplification ITGβ1-3'UTR fragments

after enzyme digestion to connect to the luciferase report carrier pMIR-Report

construct pMIR ITGβ1-3' UTR luciferase reporter gene vector and mutation carriers and identified.Will pMIR-Report Luciferase control

constructed pMIR ITGβ1-3'UTR and mutation carriers respectively with miR-mimics a total of 29 transfection to ovarian granulosa cells

with double Luciferase experimental analysis of the mechanism of action of miR-29 and ITGβ1. Result: By enzyme cutting and gene sequencing method confirmed by construction of plasmid sequence is correct;dual luciferase experiments have established that miR-29 can combine in ITGβ1-3'UTR.Successful build pMIR ITGβ1-3'UTR dual luciferase report gene

confirmed by the method of enzyme cutting and gene sequencing constructed plasmid sequence is correct.Cloning of DNA sequence fragment size and consistent with the reported in Genbank. Conclusion: The luciferase report gene build successful

miR-29 can combine after transcription ITGβ1-3'UTR.