Abstract:

Objective    To construct the eukaryotic expression vectors of Upstream Stimulatory Factor 2 (USF2) and its truncated genes，and identify the functional region of USF2 protein down regulating the Smurf1/2 transcriptional level. Methods   Using polymerase chain reaction and gene recombination techniques，USF2 gene and its two truncated genes USF2(1-235aa) and USF2(236-346aa) were amplified and cloned into the pCMV-Myc vector respectively, and then transfected into MCF7 cells to detect their expressions. The 236-346aa region at the C terminal of USF2 inhibiting the Smurf1/2 transcription was explored by Western blot and real-time PCR techniques. Results   USF2 and its truncated genes were successfully cloned into the pCMV-Myc vector. Through Western blot and real-time PCR, the 236-346aa region at the C terminal of USF2 inhibiting the transcriptional level of Smurf1/2 was proved. Conclusion   USF2 suppresses the Smurf1/2 transcriptional level through the 236-346aa region at the C terminal．

Key words: Upstream transcription factor 2; Smad ubiquitination regulator factor 1/2; Transcriptional level； Truncated genes