Characterization of porcine skeletal a-actin gene promoter: expression specificity and regulatory elements

다국어 초록 (Multilingual Abstract)

Identification of essential fragments in the promoter region of genes to drive tissue-specific expression of exogenous genes in the skeletal muscle is obligatory for animal transgenic study. The skeletal a-actin is a major protein of the thin filaments of skeletal muscle fiber. In this study, the specificity and activity of porcine skeletal aactin gene promoter were investigated by approaches of cell transfection and assayed with green fluorescent protein (GFP) and dual luciferase reporter activity, respectively.
The results revealed that the obtained 3717 bp fragment of porcine skeletal a-actin gene promoter drives GFP to be expressed uniquely in murine C2C12 cells, and two segments of 2.0–2.3 and 0.06–0.37 kb in the promoter region were essential for porcine skeletal a-actin gene expression.
The regulatory elements in the 0.06–0.37 kb fragment were further investigated, and when it is deleted or the CArG box within this fragment was mutated, the promoter activity was reduced to 20 or 43 % (P\0.01), respectively, compared to the 3.04 kb segment, suggesting an important role of CArG box in regulating porcine skeletal a-actin gene transcription. The results may provide reference for creating transgenic pigs to express exogenous genes uniquely in pork.

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