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Preliminary study on TBX1 gene promoter activity

XU Yue-juan1, PU Tian1, CAO Rui-xue1, GUO Qian-qian1, CHEN Sun1, XU Rang1,2, SUN Kun1   

  1. 1.Department of Pediatric Cardiology, 2.Scientific Research Center, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200092, China
  • Online:2013-08-28 Published:2013-09-16
  • Supported by:
    National Basic Research Program of China, “973” Program, 2010CB529501; National Natural Science Foundation of China, 81070135; Shanghai College Leading Discipline Foundation B

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Abstract:

Objective To determine the activity of TBX1 gene promoter, and identify the critical region of TBX1 gene promoter. Methods TBX1 gene promoters with
different length were isolated and amplified by PCR, digested and inserted into pGL3-Basic reporter gene vector. 293T and COS7 cells were transiently
transfected with reporter plasmids containing TBX1 gene promoters with different length using FuGene HD. Forty-two hours later, the activity of TBX1 gene
promoters with different length was compared with luciferase detection. Results There were differences in activity among TBX1 gene promoters with different
length, and the activity of TBX1 gene promoters with 5 kb and 1.2 kb in length was higher. The activity of TBX1 gene promoters varied in different cells,
with the highest activity of TBX1 gene promoter with 5 kb in length in COS7 cells and that with 1.2 kb in length in 297T cells. Conclusion In transcriptional
level, TBX1 promoter could activate the expression of reporter gene. The activity varies among promoters with different length. It is suspected that 1.2 kb
upstream region of transcript start site is critical to TBX1 gene promoter.

Key words: TBX1 gene, promoter, heart development