ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2015, Vol. 46 ›› Issue (6): 896-902.doi: 10.11843/j.issn.0366-6964.2015.06.004

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Regulatory Effects of Am80 and TSA on Chicken Stra8 Gene Promoter Activity

LIU Zhi-yong,ZUO Qi-sheng,XIAO Tian-rong,WEI Guang-hui,CHEN Ting-feng,ZHU Rui,ZHANG Zhen-tao,WANG Yi-lin,JIANG Shu-ying,ZHANG Ya-ni,LI Bi-chun*   

  1. (College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China)
  • Received:2013-12-27 Online:2015-06-23 Published:2015-06-23

Abstract:

The aims of this study were to identify the main regulatory regions of the chicken Stra8 gene promoter,to predict the binding sites of the transcription elements,and to investigate the regulatory effects of Am80 and TSA on the chicken Stra8 gene promoter activity.A series of promoter missing mutants were directly subcloned into pGL3-Basic vector,and the recombinant plasmids were transfected into DF-1 and GC-1 cells.By detecting luciferase activity and predicting binding sites of regulatory elements of promoter regulatory region,the optimal promoter fragment was selected to construct recombinant plasmid Stra8-EGFP.Am80 and TSA were applied to screen the best induced concentration,Am80 and TSA with optimal dose singly and in combination were used to induce transfected cells,and the green fluorescence expression levels in induced groups were detected.The chicken Stra8 gene promoter fragment -1 055-+54 bp had stronger activity,and had the binding sites of RARα,RXRα and RARβ;Am80 and TSA singly and in combination were applied to induce the transfected cells,and the luciferase activity was highest in the cells induced by Am80 and TSA in combination.After the cells were transfected by Stra8-EGFP,the expression of green fluorescence were the highest in cells induced by Am80(10-5 mol•L-1) and TSA(10-6 mol•L-1) in combination.The activity and binding sites of regulatory elements of chicken Stra8 gene promoter were successfully analyzed,and combined actions of Am80 and TSA significantly could promote the activity of Stra8 gene promoter.

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